59 results
Search Results
2. Decrease in Heparan Sulphate Binding in Tropism-Retargeted Oncolytic Herpes Simplex Virus (ReHV) Delays Blood Clearance and Improves Systemic Anticancer Efficacy.
- Author
-
Vannini, Andrea, Parenti, Federico, Forghieri, Cristina, Vannini, Gaia, Barboni, Catia, Zaghini, Anna, Gianni, Tatiana, and Campadelli-Fiume, Gabriella
- Subjects
TREATMENT of lung tumors ,ONCOLYTIC virotherapy ,RODENTS ,RESEARCH funding ,HERPESVIRUSES ,GLYCOPROTEINS ,TREATMENT effectiveness ,IN vivo studies ,BLOOD cells ,METASTASIS ,INTRAVENOUS therapy ,CHONDROITIN sulfates ,ENDOTHELIAL cells ,ANIMAL experimentation ,GENETIC mutation ,GLYCOSAMINOGLYCANS - Abstract
Simple Summary: Oncolytic herpes simplex viruses (oHSVs) employ as natural entry receptors nectin1 or HVEM. In contrast, retargeted oHSVs (ReHVs) employ as an entry receptor a tumor-associated antigen that no longer interacts with nectin1/HVEM and, thus, spares normal cells. Prior to entry, both oHSVs and ReHVs attach to cells by interaction of gC and gB glycoproteins with heparan sulphates and chondroitin sulphates (glycosaminoglycans–GAGs). The systemic delivery of oncolytic viruses would be the ideal route for hard-to-reach or metastatic cancers that constitute unmet clinical needs. In an accompanying paper, we reported that the blood complement and the antiviral neutralizing antibodies represent the major blood factors that inactivate systemically administered ReHVs. Here, we asked whether ReHV adsorption to GAGs acts as a sink and subtracts the virus from circulation. We report that a genetic modification in gC, which reduced its interaction with GAGs, resulted in a longer half-life of circulating ReHV and a higher anticancer efficacy of systemically (but not intratumorally) administered ReHV. The role of the interaction with cell-surface glycosaminoglycans (GAGs) during in vivo HSV infection is currently unknown. The rationale of the current investigation was to improve the anticancer efficacy of systemically administered retargeted oHSVs (ReHVs) by decreasing their binding to GAGs, including those of endothelial cells, blood cells, and off-tumor tissues. As a proof-of-principle approach, we deleted seven amino acids critical for interacting with GAGs from the glycoprotein C (gC) of R-337 ReHV. The modification in the resulting R-399 recombinant prolonged the half-life in the blood of systemically administered R-399 and enhanced its biodistribution to tumor-positive lungs and to the tumor-negative liver. Ultimately, it greatly increased the R-399 efficacy against metastatic-like lung tumors upon IV administration but not against subcutaneous tumors upon IT administration. These results provide evidence that the increased efficacy seen upon R-399 systemic administration correlated with the slower clearance from the circulation. To our knowledge, this is the first in vivo evidence that the partial impairment of the gC interaction with GAGs resulted in a prolonged half-life of circulating ReHV, an increase in the amount of ReHV taken up by tissues and tumors, and, ultimately, an enhanced anticancer efficacy of systemically administered ReHV. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
3. Derivatives of Betulin and Betulinic Acid Containing a Phosphonate Group—In Silico Studies and Preliminary In Vitro Assessment of Antiviral Activity.
- Author
-
Bębenek, Ewa, Pęcak, Paweł, Kadela-Tomanek, Monika, Orzechowska, Beata, and Chrobak, Elwira
- Subjects
BETULINIC acid ,BETULIN ,ANTIVIRAL agents ,PHOSPHONATES ,MOLECULAR docking ,VIRUS diseases ,ANIMAL experimentation - Abstract
Viral diseases affecting both humans and animals are a serious public problem. Chemical modifications of the structure of compounds of natural origin, e.g., betulin, seem to be a promising model in the search for new antiviral agents. The subject of our work was to conduct preliminary tests on the antiviral activity of phosphonic derivatives of betulin and betulinic acid and to assess the pharmacokinetic profile of target compounds. Human (HHV-1, HAdV-5) and animal viruses (BEV, VSV) were used in the in vitro tests. Additionally, this paper presents the results of research using in silico methods (ADMET and molecular docking). Two compounds (betulin 29-phosphonate 3 and 3-(3′,3′-dimethylsuccinyl)betulin acid 29-phosphonate 8a) showed antiviral activity against BEV, and compound 3 was also active against HAdV-5. For compound 3, which showed advantageous pharmacokinetic parameters, molecular docking was performed to determine possible interactions with the cellular target HAdV-5 endopeptidase, which plays an important role in various functions of the virus. Selecting the most active derivatives makes it possible to plan tests on an animal model. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
4. Triptolide Reduces Neoplastic Progression in Hepatocellular Carcinoma by Downregulating the Lipid Lipase Signaling Pathway.
- Author
-
Chang, Wei, Wang, Jingjing, You, Yuanqi, Wang, Hongqian, Xu, Shendong, Vulcano, Stephen, Xu, Changlu, Shen, Chenlin, Li, Zhi, and Wang, Jie
- Subjects
LIPID metabolism ,LIPASES ,EXPERIMENTAL design ,TERPENES ,CELL migration ,ANALYSIS of variance ,CARCINOGENESIS ,ANTI-inflammatory agents ,ANIMAL experimentation ,MICROBIOLOGICAL assay ,IMMUNOHISTOCHEMISTRY ,APOPTOSIS ,CELLULAR signal transduction ,RESEARCH funding ,CELL lines ,POLYMERASE chain reaction ,DATA analysis software ,HEPATOCELLULAR carcinoma ,MICE ,PHARMACODYNAMICS ,DISEASE risk factors - Abstract
Simple Summary: TP is a widely utilized anticancer medication, particularly effective in treating HCC. Thorough investigation into TP's molecular mechanism in HCC treatment is crucial for precise and combination therapy. In this paper, we have unveiled a novel mechanism of TP in HCC treatment, where it inhibits tumor growth by reducing lipid accumulation. While the inhibition of P53 gene activity is commonly thought to be the reason for TP's effectiveness in treating HCC, this new mechanism serves as a significant complement to the current understanding, paving the way for innovative approaches to HCC treatment. Hepatocellular carcinoma (HCC), which is the third leading cause of cancer-related mortality in the world, presents a significant medical challenge. Triptolide (TP) has been identified as an effective therapeutic drug for HCC. However, its precise therapeutic mechanism is still unknown. Understanding the mechanism of action of TP against HCC is crucial for its implementation in the field of HCC treatment. We hypothesize that the anti-HCC actions of TP might be related to its modulation of HCC lipid metabolism given the crucial role that lipid metabolism plays in promoting the progression of HCC. In this work, we first demonstrate that, both in vitro and in vivo, TP significantly reduces lipid accumulation in HCC cells. Additionally, we notice that lipoprotein lipase (LPL) expression is markedly upregulated in HCC, and that its levels are positively connected with the disease's progression. It is interesting to note that TP dramatically reduces LPL activity, which in turn prevents HCC growth and reduces lipid accumulation. Additionally, the effect of TP on LPL is a direct correlation. These results definitely demonstrate that TP protects hepatocytes against abnormal accumulation of lipids by transcriptionally suppressing LPL, which reduces the development of HCC. This newly identified pathway provides insight into the process through which TP exerts its anti-HCC actions. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
5. Continuous Electrode Models and Application of Exact Schemes in Modeling of Electrical Impedance Measurements.
- Author
-
Vizvari, Zoltan, Klincsik, Mihaly, Odry, Peter, Tadic, Vladimir, Gyorfi, Nina, Toth, Attila, and Sari, Zoltan
- Subjects
ELECTRIC impedance ,ELECTRODES ,ANIMAL experimentation ,CONTINUOUS functions ,POTENTIAL functions ,DIFFERENTIAL equations - Abstract
The crucial issue in electrical impedance (EI) measurements lies in the galvanic interaction between the electrodes and the investigated material. This paper brings together the basic and applied research experience and combines their results with excellent properties. Consequently, innovative precise methodologies have emerged, enabling the direct modeling of EI measurements, free from the inaccuracies often associated with numerical approaches. As an outcome of the efficiency and robustness of the applied method, the conductivity of the material and the electrodes are represented by a common piecewise function, which is used to solve the differential equation modeling of the EI measurement. Moreover, this allows the possibility for modeling the conductivity of electrodes with continuous functions, providing an important generalization of the Complete Electrode Model (CEM), which has been widely used so far. The effectiveness of the novel approach was showcased through two distinct case studies. In the first case study, potential functions within both the material and the electrodes were computed using the CEM. In the second case study, calculations were performed utilizing the newly introduced continuous electrode model. The simulation results suggest that the new method is a powerful tool for biological research, from in vitro experiments to animal studies and human applications. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
6. Toward Transparency on Animal Experimentation in Switzerland: Seven Recommendations for the Provision of Public Information in Swiss Law.
- Author
-
Lüthi, Nicole, Rodriguez Perez, Christian, Persson, Kirsten, Elger, Bernice Simone, and Shaw, David
- Subjects
ANIMAL welfare laws ,ANIMAL experimentation ,ANIMAL breeding ,SCIENTIFIC community ,ANIMAL breeds - Abstract
Simple Summary: Simple Summary: In Switzerland, the importance of transparency in animal experimentation is emphasized by the Swiss Federal Council, in recognition of the public's great interest in this matter. Current Swiss law requires institutions and the government to inform the public about various aspects, such as the number of animals used in experiments, their species, or the severity of harm of the experiment. However, much relevant information is missing, such as the number of breeding or surplus animals, the fate of the animals (both in facilities and after an experiment), and how the balancing of animal harm and human benefits has been performed to justify a particular experiment. Considering that the Swiss government has a duty to provide information on animal experimentation conducted on the public's behalf, such information should be disclosed. If Switzerland is to move toward more transparency in public information on animal experimentation, an update of the legal requirements is needed. In this article, we give recommendations for Swiss law to move toward more transparency in public information. In Switzerland, the importance of transparency in animal experimentation is emphasized by the Swiss Federal Council, recognizing the public's great interest in this matter. Federal reporting on animal experimentation indicates a total of 585,991 animals used in experiments in Switzerland in 2022. By Swiss law, the report enables the public to learn about many aspects such as the species and degree of suffering experienced by the animals, but some information of interest to the public is missing, such as the fate of the animals at the end of the experiment (e.g., euthanized, rehomed in a private home, reused in another experiment). When it comes to animals bred in facilities but not used in experiments, further information of interest is not required to be made public according to Swiss law, for example, the number and fate of "surplus" animals (i.e., animals bred but not used in experiments for a variety of reasons such as not carrying the phenotypical properties needed). Considering that the Swiss government has a duty to provide a full accounting of animal experimentation conducted on the public's behalf, further relevant information should be disclosed. While efforts toward transparency, such as the STAAR Agreement, have been made in the scientific community, these mostly reflect the legal requirements already in force. If Switzerland is to move toward more transparency in public information on animal experimentation, an update of the legal requirements is needed. In this article, we give recommendations for Swiss law to move toward more transparency in public information on seven aspects: (1) the fate of the animals at the end of the experiment; (2) the sources of funding for animal experimentation; (3) the harm-benefit analysis performed by researchers and ethics committees to justify an experiment using animals; (4) the number of breeding/surplus animals; (5) the fate of breeding/surplus animals; (6) the harms experienced by animals in facilities; and (7) the funding of animal facilities. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
7. Evaluation of the Mammalian Aquaporin Inhibitors Auphen and Z433927330 in Treating Breast Cancer.
- Author
-
Charlestin, Verodia, Tan, Elijah, Arias-Matus, Carlos Eduardo, Wu, Junmin, Miranda-Vergara, Maria Cristina, Lee, Mijoon, Wang, Man, Nannapaneni, Dharma T., Tennakoon, Parinda, Blagg, Brian S. J., Ashfeld, Brandon L., Kaliney, William, Li, Jun, and Littlepage, Laurie E.
- Subjects
THERAPEUTIC use of antineoplastic agents ,PROTEINS ,CARRIER proteins ,CANCER invasiveness ,GOLD compounds ,MITOCHONDRIA ,RESEARCH funding ,RECEIVER operating characteristic curves ,BREAST tumors ,ANTINEOPLASTIC agents ,CELL physiology ,POLYMERASE chain reaction ,TRANSCRIPTION factors ,TAMOXIFEN ,DESCRIPTIVE statistics ,METASTASIS ,GENE expression ,MICE ,CELL lines ,EXPERIMENTAL design ,RNA ,IMMUNOHISTOCHEMISTRY ,ANIMAL experimentation ,MOLECULAR structure ,WESTERN immunoblotting ,DRUG efficacy ,CELL survival ,OVERALL survival ,PHARMACODYNAMICS ,CHEMICAL inhibitors - Abstract
Simple Summary: Aquaporins (AQPs) have emerged as potential predictors of response to cancer therapy and as targets for increasing sensitivity to treatment. However, systemic therapeutic inhibition using pan-AQP or AQP-specific inhibitors has not been characterized for efficacy in treating breast cancer or as a component of combination treatment. We evaluated AQP inhibition using established AQP inhibitors in cytotoxicity and pharmacologic assays. This study identified AQPs as a targetable vulnerability in breast cancer. AQP inhibitors can increase therapeutic efficacy, both as single agents and in a combination therapy strategy, in treating breast cancer progression and metastasis. These results provide significant insights that support the future development of improved AQP inhibitors. AQPs contribute to breast cancer progression and metastasis. We previously found that genetic inhibition of Aqp7 reduces primary tumor burden and metastasis in breast cancer. In this study, we utilized two AQP inhibitors, Auphen and Z433927330, to evaluate the efficacy of therapeutic inhibition of AQPs in breast cancer treatment. The inhibitors were evaluated in breast cancer for both cytotoxicity and metabolic stability assays across both murine and human breast cancer cell lines. Both AQP inhibitors also affected the expression of other AQP transcripts and proteins, which demonstrates compensatory regulation between AQP family members. As a single agent, Auphen treatment in vivo extended overall survival but did not impact primary or metastatic tumor burden. However, Auphen treatment made cells more responsive to chemotherapy (doxorubicin) or endocrine treatment (tamoxifen, fulvestrant). In fact, treatment with Tamoxifen reduced overall AQP7 protein expression. RNA-seq of breast cancer cells treated with Auphen identified mitochondrial metabolism genes as impacted by Auphen and may contribute to reducing mammary tumor progression, lung metastasis, and increased therapeutic efficacy of endocrine therapy in breast cancer. Interestingly, we found that Auphen and tamoxifen cooperate to reduce breast cancer cell viability, which suggests that Auphen treatment makes the cells more susceptible to Tamoxifen. Together, this study highlights AQPs as therapeutic vulnerabilities of breast cancer metastasis that are promising and should be exploited. However, the pharmacologic results suggest additional chemical refinements and optimization of AQP inhibition are needed to make these AQP inhibitors appropriate to use for therapeutic benefit in overcoming endocrine therapy resistance. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
8. Neuro-Oncologic Veterinary Trial for the Clinical Transfer of Microbeam Radiation Therapy: Acute to Subacute Radiotolerance after Brain Tumor Irradiation in Pet Dogs.
- Author
-
Eling, Laura, Kefs, Samy, Keshmiri, Sarvenaz, Balosso, Jacques, Calvet, Susan, Chamel, Gabriel, Drevon-Gaud, Renaud, Flandin, Isabelle, Gaudin, Maxime, Giraud, Lucile, Laissue, Jean Albert, Pellicioli, Paolo, Verry, Camille, Adam, Jean-François, and Serduc, Raphaël
- Subjects
HETEROCYCLIC compounds ,GLIOMAS ,RADIOTHERAPY ,PETS ,RESEARCH funding ,DATA analysis ,T-test (Statistics) ,BRAIN ,QUESTIONNAIRES ,PHENOBARBITAL ,DOGS ,MAGNETIC resonance imaging ,CANCER patients ,DESCRIPTIVE statistics ,TREATMENT effectiveness ,RADIATION dosimetry ,PREDNISOLONE ,ANIMAL experimentation ,QUALITY of life ,SEIZURES (Medicine) ,STATISTICS ,RADIATION doses ,DATA analysis software ,BRAIN tumors - Abstract
Simple Summary: The benefit of spatially fractionated radiotherapy for brain tumors is maximized through Synchrotron Microbeam Radiation Therapy (MRT). In 2021, the clinical transfer phase of MRT began: the first brain-tumor-bearing dog patients were treated under clinical conditions in view of the forthcoming clinical transfer. As a primary endpoint, the tolerance of normal brain tissues to MRT was evaluated, while the efficacy in reducing tumor volume was considered as a secondary endpoint. We here present acute to subacute neurologic radiotolerance and tumor volume reduction after MRT for brain tumor treatment in canine patients included in our ongoing veterinary trial, proving that MRT is a safe tool for spontaneous brain tumor treatment in dogs. Synchrotron Microbeam Radiation Therapy (MRT) has repeatedly proven its superiority compared with conventional radiotherapy for glioma control in preclinical research. The clinical transfer phase of MRT has recently gained momentum; seven dogs with suspected glioma were treated under clinical conditions to determine the feasibility and safety of MRT. We administered a single fraction of 3D-conformal, image-guided MRT. Ultra-high-dose rate synchrotron X-ray microbeams (50 µm-wide, 400 µm-spaced) were delivered through five conformal irradiation ports. The PTV received ~25 Gy peak dose (within microbeams) per port, corresponding to a minimal cumulated valley dose (diffusing between microbeams) of 2.8 Gy. The dogs underwent clinical and MRI follow-up, and owner evaluations. One dog was lost to follow-up. Clinical exams of the remaining six dogs during the first 3 months did not indicate radiotoxicity induced by MRT. Quality of life improved from 7.3/10 [±0.7] to 8.9/10 [±0.3]. Tumor-induced seizure activity decreased significantly. A significant tumor volume reduction of 69% [±6%] was reached 3 months after MRT. Our study is the first neuro-oncologic veterinary trial of 3D-conformal Synchrotron MRT and reveals that MRT does not induce acute to subacute radiotoxicity in normal brain tissues. MRT improves quality of life and leads to remarkable tumor volume reduction despite low valley dose delivery. This trial is an essential step towards the forthcoming clinical application of MRT against deep-seated human brain tumors. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
9. Radiosynthesis and Preclinical Evaluation of 18 F-Labeled Estradiol Derivatives with Different Lipophilicity for PET Imaging of Breast Cancer.
- Author
-
Friedel, Anna, Prante, Olaf, and Maschauer, Simone
- Subjects
BREAST tumor diagnosis ,CLINICAL drug trials ,BIOLOGICAL models ,IN vitro studies ,RADIOPHARMACEUTICALS ,RESEARCH funding ,BREAST tumors ,ANTINEOPLASTIC agents ,POSITRON emission tomography ,TREATMENT effectiveness ,IN vivo studies ,CANCER cell culture ,RADIOISOTOPES ,DESCRIPTIVE statistics ,ESTRADIOL ,MICE ,CELL lines ,ANIMAL experimentation ,MOLECULAR structure ,FLUORINE isotopes ,DRUG development ,COMPARATIVE studies ,PHARMACODYNAMICS - Abstract
Simple Summary: Breast cancer is one of the most prevalent forms of cancer diagnosed in women worldwide. Since the estradiol receptor (ER) is overexpressed in 75% of breast tumors, it is a reasonable target for tumor diagnosis and therapy. This study focuses on the development and preclinical evaluation of readily synthesized
18 F-labeled estradiol derivatives with different lipophilicity. The least hydrophilic derivative,18 F-TA-Glyco-EE, showed the highest cellular uptake in ER-positive breast cancer cells. The in vivo PET imaging of breast tumor-bearing mice demonstrated the desired rapid clearance of the tracer from the excretory organ through the liver. The in vitro autoradiography of ER-positive tumor sections confirmed the high specific binding of18 F-TA-Glyco-EE. In conclusion,18 F-TA-Glyco-EE may be a promising candidate for imaging of ER-positive breast cancer. About 75% of breast tumors show an overexpression of the estradiol receptor (ER), making it a valuable target for tumor diagnosis and therapy. To date, 16α-[18 F]fluoroestradiol (FES) is the only FDA-approved imaging probe for the positron emission tomography (PET) imaging of ER-positive (ER+) breast cancer. However, FES has the drawback of a high retention in the liver. Therefore, the aim of this study was the development and preclinical evaluation of estradiol (E2) derivatives with different lipophilicity. Three18 F-labeled prosthetic groups (two glycosyl and one PEG azide) were chosen for conjugation with ethinyl estradiol (EE) by18 F-CuAAC (Cu-catalyzed azide-alkyne cycloaddition). The cellular uptake in ER+ MCF-7 tumor cells was highest for the less hydrophilic derivative (18 F-TA-Glyco-EE). In nude mice bearing different breast tumors (ER+ MCF-7 and T47D versus ER− MDA-MB-231),18 F-TA-Glyco-EE revealed a high uptake in the liver (13%ID/g, 30 min p.i.), which decreased over 90 min to 1.2%ID/g, indicating fast hepatobiliary clearance. The statistically significant difference of18 F-TA-Glyco-EE uptake in T47D compared to MDA-MB-231 tumors at 60–90 min p.i. indicated ER-specific uptake, whereas in vivo PET imaging did not provide evidence for specific uptake of18 F-TA-Glyco-EE in MCF-7 tumors, probably due to ER occupation by E2 after E2-dependent MCF-7 tumor growth in mice. However, in vitro autoradiography revealed a high specific binding of18 F-TA-Glyco-EE to ER+ tumor slices. We conclude that18 F-TA-Glyco-EE, with its increased hydrophilicity after deacetylation in the blood and thus rapid washout from non-target tissues, may be a viable alternative to FES for the PET imaging of breast cancer. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
10. Characterization of a Syngeneic Orthotopic Model of Cholangiocarcinoma by [ 18 F]FDG-PET/MRI.
- Author
-
Zachhuber, Lena, Filip, Thomas, Mozayani, Behrang, Löbsch, Mathilde, Scheiner, Stefan, Vician, Petra, Stanek, Johann, Hacker, Marcus, Helbich, Thomas H., Wanek, Thomas, Berger, Walter, and Kuntner, Claudia
- Subjects
BIOLOGICAL models ,LIVER tumors ,RADIOPHARMACEUTICALS ,RESEARCH funding ,CHOLANGIOCARCINOMA ,DEOXY sugars ,POSITRON emission tomography ,MAGNETIC resonance imaging ,MICE ,METASTASIS ,BLOOD sugar ,ANIMAL experimentation - Abstract
Simple Summary: Cholangiocarcinoma (CCA) is a type of liver cancer with few treatment options and low survival rates in advanced stages. Our study developed a mouse model to study this cancer type by implanting CCA cells into the liver of mice. We used advanced imaging techniques (MRI and PET scans) to monitor tumor growth and metabolism over four weeks. We observed that tumors became visible early and grew steadily over time. PET scans showed increasing tumor activity, and blood tests revealed liver damage. Most mice developed lung metastases after four weeks. Our research shows that combining MRI and PET scans effectively tracks CCA progression in mice, providing valuable insights into cancer development and investigating potential treatments. Cholangiocarcinoma (CCA) is a type of primary liver cancer originating from the biliary tract epithelium, characterized by limited treatment options for advanced cases and low survival rates. This study aimed to establish an orthotopic mouse model for CCA and monitor tumor growth using PET/MR imaging. Murine CCA cells were implanted into the liver lobe of male C57BL/6J mice. The imaging groups included contrast-enhanced (CE) MR, CE-MR with static [
18 F]FDG-PET, and dynamic [18 F]FDG-PET. Tumor volume and FDG uptake were measured weekly over four weeks. Early tumor formation was visible in CE-MR images, with a gradual increase in volume over time. Dynamic FDG-PET revealed an increase in the metabolic glucose rate (MRGlu) over time. Blood analysis showed pathological changes in liver-related parameters. Lung metastases were observed in nearly all animals after four weeks. The study concludes that PET-MR imaging effectively monitors tumor progression in the CCA mouse model, providing insights into CCA development and potential treatment strategies. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
11. Exploring Canine Mammary Cancer through Liquid Biopsy: Proteomic Profiling of Small Extracellular Vesicles.
- Author
-
Novais, Adriana Alonso, Tamarindo, Guilherme Henrique, Melo, Luryan Mikaelly Minotti, Balieiro, Beatriz Castilho, Nóbrega, Daniela, dos Santos, Gislaine, Saldanha, Schaienni Fontoura, de Souza, Fabiana Ferreira, Chuffa, Luiz Gustavo de Almeida, Bracha, Shay, and Zuccari, Debora Aparecida Pires de Campos
- Subjects
BREAST cancer prognosis ,BREAST tumor diagnosis ,PROTEIN analysis ,EXTRACELLULAR vesicles ,BIOLOGICAL models ,CANCER relapse ,CENTRIFUGATION ,LIQUID chromatography-mass spectrometry ,RESEARCH funding ,ELECTRON microscopy ,BODY fluid examination ,TUMOR markers ,DOGS ,DISEASE remission ,DESCRIPTIVE statistics ,PROTEOMICS ,ANIMAL experimentation ,MASS spectrometry ,GENE expression profiling ,RESEARCH methodology ,EXOSOMES - Abstract
Simple Summary: We studied canine mammary tumors to better understand similar human breast cancer using a technique called liquid biopsy, which analyzes blood samples to detect disease, focusing on the detection of tiny particles called small extracellular vesicles. These structures are very interesting because they can carry proteins that may indicate the presence of cancer. In this study, we collected blood from healthy dogs, dogs with benign and malignant CMTs, and those in remission and also with recurrence. We found no differences in the size or amount of the vesicles among the groups but identified specific proteins that could serve as markers for cancer. These proteins could potentially help in the diagnosis, prognosis and monitoring of mammary cancer. (Background). Canine mammary tumors (CMTs) have emerged as an important model for understanding pathophysiological aspects of human disease. Liquid biopsy (LB), which relies on blood-borne biomarkers and offers minimal invasiveness, holds promise for reflecting the disease status of patients. Small extracellular vesicles (SEVs) and their protein cargo have recently gained attention as potential tools for disease screening and monitoring. (Objectives). This study aimed to isolate SEVs from canine patients and analyze their proteomic profile to assess their diagnostic and prognostic potential. (Methods). Plasma samples were collected from female dogs grouped into CMT (malignant and benign), healthy controls, relapse, and remission groups. SEVs were isolated and characterized using ultracentrifugation (UC), nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Proteomic analysis of circulating SEVs was conducted using liquid chromatography–mass spectrometry (LC–MS). (Results). While no significant differences were observed in the concentration and size of exosomes among the studied groups, proteomic profiling revealed important variations. Mass spectrometry identified exclusive proteins that could serve as potential biomarkers for mammary cancer. These included Inter-alpha-trypsin inhibitor heavy chain (ITIH2 and ITI4), phosphopyruvate hydratase or alpha enolase (ENO1), eukaryotic translation elongation factor 2 (eEF2), actin (ACTB), transthyretin (TTR), beta-2-glycoprotein 1 (APOH) and gelsolin (GSN) found in female dogs with malignant tumors. Additionally, vitamin D-binding protein (VDBP), also known as group-specific component (GC), was identified as a protein present during remission. (Conclusions). The results underscore the potential of proteins found in SEVs as valuable biomarkers in CMTs. Despite the lack of differences in vesicle concentration and size between the groups, the analysis of protein content revealed promising markers with potential applications in CMT diagnosis and monitoring. These findings suggest a novel approach in the development of more precise and effective diagnostic tools for this challenging clinical condition. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
12. Glioma-Stem-Cell-Derived Exosomes Remodeled Glioma-Associated Macrophage via NEAT1/miR-125a/STAT3 Pathway.
- Author
-
Pan, Tong, Xie, Dong-Kun, Li, Juan, Qiang, Yu-Jie, Fan, Song-Yuan, Wang, Ting-Ting, Han, Yuan-Yuan, Zang, Jian, Yang, Yang, Zhao, Jun-Long, Li, San-Zhong, and Wu, Shuang
- Subjects
GLIOMA treatment ,GLIOMAS ,MACROPHAGES ,RADIOTHERAPY ,DRUG resistance in cancer cells ,RESEARCH funding ,MICRORNA ,CELLULAR signal transduction ,MICE ,CANCER chemotherapy ,ANIMAL experimentation ,DNA damage ,STEM cells ,STAT proteins ,EXOSOMES ,IMMUNOSUPPRESSION ,DISEASE progression - Abstract
Simple Summary: Glioblastomas (GBMs) are considered the most lethal cancer in the central nervous system (CNS), whose malignant phenotypes are majorly attributed to glioma stem cells (GSCs). Despite combined surgical radiotherapy with temozolomide chemotherapy and tumor-treating fields (TTFs), the tumor almost always recurs near the resection site. Besides the contribution of GSCs, the tumor microenvironment (TME) also plays an important role in glioma recurrence. Our work has demonstrated that GSC-derived exosomes carry lncRNA NEAT1 to promote the M2 polarization of glioma-associated macrophages (GAMs). Further mechanism exploration indicated that NEAT1 represses the expression of miR-125a in GAMs significantly. The decrease in miR-125a induces the elevation of target gene STAT3, which is required for macrophage M2 polarization. The development of M2-like GAMs contributes to the immunosuppressive microenvironment and glioma progression. Our findings elucidate the functions and mechanisms of the crosstalk between GSCs and GAMs via exosomes, providing new therapeutic targets and strategies for glioma. Glioblastoma (GBM), as the most common primary brain tumor, usually results in an extremely poor prognosis, in which glioma stem cells (GSCs) and their immunosuppressive microenvironment prominently intervene in the resistance to radiotherapy and chemotherapy that directly leads to tumor recurrence and shortened survival time. The specific mechanism through which exosomes generated from GSCs support the creation of an immunosuppressive microenvironment remains unknown, while it is acknowledged to be engaged in intercellular communication and the regulation of the glioma immunosuppressive microenvironment. The elevated expression of LncRNA-NEAT1 was found in glioma cells after radiotherapy, chemotherapy, and DNA damage stimulation, and NEAT1 could promote the malignant biological activities of GSCs. Emerging evidence suggests that lncRNAs may reply to external stimuli or DNA damage by playing a role in modulating different aspects of tumor biology. Our study demonstrated a promotive role of the carried NEAT1 by GSC-derived exosomes in the polarization of M2-like macrophages. Further experiments demonstrated the mediative role of miR-125a and its target gene STAT3 in NEAT1-induced polarization of M2-like macrophages that promote glioma progression. Our findings elucidate the mechanism by which GSCs influence the polarization of M2-like macrophages through exosomes, which may contribute to the formation of immunosuppressive microenvironments. Taken together, our study reveals the miR-125a-STAT3 pathway through which exosomal NEAT1 from treatment-resistant GSCs contributes to M2-like macrophage polarization, indicating the potential of exosomal NEAT1 for treating glioma. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
13. Modulating Tumor Immunity by Targeting Tumor Fibrotic Stroma and Angiogenic Vessels for Lung Cancer Treatment.
- Author
-
Yuan, Yi, Mishra, Falguni, Li, Bin, Peng, Guangda, Chan, Payton, Yang, Jenny, and Liu, Zhiren
- Subjects
TREATMENT of lung tumors ,VASCULAR endothelial growth factors ,CANCER ,MACROPHAGES ,RESEARCH funding ,IMMUNOTHERAPY ,APOPTOSIS ,PROGRAMMED death-ligand 1 ,CANCER cell culture ,DESCRIPTIVE statistics ,IMMUNE checkpoint inhibitors ,FIBROBLASTS ,MICE ,LUNG tumors ,ANIMAL experimentation ,ONE-way analysis of variance ,DATA analysis software ,SURVIVAL analysis (Biometry) - Abstract
Simple Summary: Yuan et al. present a strategy to modulate tumor immunity by simultaneously depleting CAFs and tumor angiogenic vessels using a rationally designed protein that induces integrin α
v β3 -expressing cell apoptosis. The study offers a unique opportunity for the enhancement of cancer immunotherapies, especially for patients with a tumor of dense stroma and high angiogenesis. Fibrotic stroma and angiogenic tumor vessels play an important role in modulating tumor immunity. We previously reported a rationally designed protein (ProAgio) that targets integrin αv β3 at a novel site. ProAgio induces the apoptosis of cells that express high levels of the integrin. Both activated cancer-associated fibroblasts (CAFs) and angiogenic endothelial cells (aECs) in tumors express high levels of integrin αv β3 . ProAgio simultaneously and specifically induces apoptosis in CAFs and aECs in tumors. We provide evidence here that the depletion of CAFs and the elimination of leaky tumor angiogenic vessels by ProAgio alter tumor immunity. ProAgio reduces CD4 + Treg and Myeloid-derived suppressor cells (MDSCs), increases CD8 + T-cells, and increases the M1/M2 macrophage ratio in the tumor. The depletion of dense fibrotic stroma (CAFs) by ProAgio decreases the Programmed Death Ligand 1 (PDL-1) levels in the stroma areas surrounding the tumors, and thus strongly increases the delivery of anti-PDL-1 antibody to the target cancer cells. The impact of ProAgio on tumor immunity provides strong synergistical effects of checkpoint inhibitors on lung cancer treatment. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
14. Monte Carlo-Based Nanoscale Dosimetry Holds Promise for Radiopharmaceutical Therapy Involving Auger Electron Emitters.
- Author
-
Kwon, Ohyun, Hoffman, Sabrina L. V., Ellison, Paul A., and Bednarz, Bryan P.
- Subjects
BIOLOGICAL models ,RADIOTHERAPY ,IN vivo toxicity testing ,RADIOPHARMACEUTICALS ,PREDICTION models ,RESEARCH funding ,BENCHMARKING (Management) ,RADIATION dosimetry ,ELECTRONS ,DNA ,DESCRIPTIVE statistics ,MICE ,DOSE-effect relationship in pharmacology ,SIMULATION methods in education ,DNA damage ,ANIMAL experimentation ,MOLECULAR structure ,TUMORS ,COMPARATIVE studies ,DATA analysis software ,GENETICS - Abstract
Simple Summary: In contrast to external beam radiation therapy, our understanding of the dose–response relationship in radiopharmaceutical therapy (RPT) remains limited. Given that the estimation of radiation-induced deoxyribonucleic acid (DNA) damage can correlate with various biological responses, nanoscale dosimetry emerges as a critical tool in lighting this intricate relationship. In this study, we developed a novel simulation-based nanoscale dosimetry platform by identifying optimal DNA physics models and benchmarking against existing data. Leveraging this platform, we conducted a case study to estimate DNA damage, thereby enhancing our comprehension of RPT effects at nanoscales. This research serves as a foundation for future studies on how RPT affects DNA at the nano-level, potentially improving cancer treatment strategies. Radiopharmaceutical therapy (RPT) is evolving as a promising strategy for treating cancer. As interest grows in short-range particles, like Auger electrons, understanding the dose–response relationship at the deoxyribonucleic acid (DNA) level has become essential. In this study, we used the Geant4-DNA toolkit to evaluate DNA damage caused by the Auger-electron-emitting isotope I-125. We compared the energy deposition and single strand break (SSB) yield at each base pair location in a short B-form DNA (B-DNA) geometry with existing simulation and experimental data, considering both physical direct and chemical indirect hits. Additionally, we evaluated dosimetric differences between our high-resolution B-DNA target and a previously published simple B-DNA geometry. Overall, our benchmarking results for SSB yield from I-125 decay exhibited good agreement with both simulation and experimental data. Using this simulation, we then evaluated the SSB and double strand break (DSB) yields caused by a theranostic Br-77-labeled poly ADP ribose polymerase (PARP) inhibitor radiopharmaceutical. The results indicated a predominant contribution of chemical indirect hits over physical direct hits in generating SSB and DSB. This study lays the foundation for future investigations into the nano-dosimetric properties of RPT. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
15. Screening of Secretory Proteins Linking Major Depressive Disorder with Heart Failure Based on Comprehensive Bioinformatics Analysis and Machine Learning.
- Author
-
Zhang, Chuanjing, Song, Yongfei, Cen, Lichao, Huang, Chen, Zhou, Jianqing, and Lian, Jiangfang
- Subjects
MENTAL depression ,VENTRICULAR ejection fraction ,ANIMAL experimentation ,HEART failure ,T cells - Abstract
Background: Major depressive disorder (MDD) plays a crucial role in the occurrence of heart failure (HF). This investigation was undertaken to explore the possible mechanism of MDD's involvement in HF pathogenesis and identify candidate biomarkers for the diagnosis of MDD with HF. Methods: GWAS data for MDD and HF were collected, and Mendelian randomization (MR) analysis was performed to investigate the causal relationship between MDD and HF. Differential expression analysis (DEA) and WGCNA were used to detect HF key genes and MDD-associated secretory proteins. Protein–protein interaction (PPI), functional enrichment, and cMAP analysis were used to reveal potential mechanisms and drugs for MDD-related HF. Then, four machine learning (ML) algorithms (including GLM, RF, SVM, and XGB) were used to screen candidate biomarkers, construct diagnostic nomograms, and predict MDD-related HF. Furthermore, the MCPcounter algorithm was used to explore immune cell infiltration in HF, and MR analysis was performed to explore the causal effect of immunophenotypes on HF. Finally, the validation of the association of MDD with reduced left ventricular ejection fraction (LVEF) and the performance assessment of diagnostic biomarkers was accomplished based on animal models mimicking MDD. Results: The MR analysis showed that the MDD was linked to an increased risk of HF (OR = 1.129, p < 0.001). DEA combined with WGCNA and secretory protein gene set identified 315 HF key genes and 332 MDD-associated secretory proteins, respectively. Through PPI and MCODE analysis, 78 genes were pinpointed as MDD-related pathogenic genes for HF. The enrichment analysis revealed that these genes were predominantly enriched in immune and inflammatory regulation. Through four ML algorithms, two hub genes (ISLR/SFRP4) were identified as candidate HF biomarkers, and a nomogram was developed. ROC analysis showed that the AUC of the nomogram was higher than 0.90 in both the HF combined dataset and two external cohorts. In addition, an immune cell infiltration analysis revealed the immune dysregulation in HF, with ISLR/SFRP4 displaying notable associations with the infiltration of B cells, CD8 T cells, and fibroblasts. More importantly, animal experiments showed that the expression levels of ISLR (r = −0.653, p < 0.001) and SFRP4 (r = −0.476, p = 0.008) were significantly negatively correlated with LVEF. Conclusions: The MR analysis indicated that MDD is a risk factor for HF at the genetic level. Bioinformatics analysis and the ML results suggest that ISLR and SFRP4 have the potential to serve as diagnostic biomarkers for HF. Animal experiments showed a negative correlation between the serum levels of ISLR/SFRP4 and LVEF, emphasizing the need for additional clinical studies to elucidate their diagnostic value. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
16. Development and Evaluation of Bioconverted Milk with Anti-Microbial Effect against Periodontal Pathogens and α-Glucosidase Inhibitory Activity.
- Author
-
Lee, Yewon, Yoon, Yohan, and Choi, Kyoung-Hee
- Subjects
LACTIC acid bacteria ,AMINO acid metabolism ,ACTINOBACILLUS actinomycetemcomitans ,GLYCEMIC control ,ANIMAL experimentation - Abstract
To decrease periodontal pathogens and increase the number of beneficial bacteria, probiotics and bioactive compounds made via microbial bioconversion are recently used. In addition, the interest regarding probiotics-mediated bioconversion with popular medicinal plants is increasing. Artemisia herba-alba, a type of wormwood, has recently been attention as a medicinal plant due to its various bioactive compounds. Therefore, we developed bioconverted milk containing A. herba-alba that effectively inhibited periodontal pathogens and α-glucosidase. To select the appropriate lactic acid bacteria for the probiotic candidate strain, 74 strains of lactic acid bacteria were screened. Among them, Lactiplantibacillus plantarum SMFM2016-RK was chosen as the probiotic due to its beneficial characteristics such as high acid and bile tolerance, antioxidant activity, and α-glucosidase inhibition. Based on the minimal bactericidal concentration against three periodontal pathogens, the following appropriate concentrations of Artemisia herba-alba extract were added to milk: 5 mg/mL of A. herba-alba ethanol extract and 25 mg/mL of A. herba-alba hot-water extract. Four bioconverted milks (BM), BM1, BM2, BM3, and BM4, were produced by combining L. plantarum SMFM2016-RK alone, L. plantarum SMFM2016-RK and ethanol extract, L. plantarum SMFM2016-RK and hot-water extract, and L. plantarum SMFM2016-RK with both extracts. As a result of antimicrobial activity, BM3 inhibited the growth of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis the most, and BM4 suppressed the growth of Fusobacterium nucleatum the most. In addition, bioconverted milk containing A. herba-alba (BM2, BM3, and BM4) inhibited α-glucosidase more effectively than BM1. The whole genome of L. plantarum SMFM2016-RK was obtained, and 3135 CDS, 67 tRNA, and 16 RNA were predicted. The genome annotation of L. plantarum SMFM2016-RK revealed 11 CDS related to proteolysis and amino acid metabolism and 2 CDS of phenolic acid-metabolizing enzymes. In conclusion, A. herba-alba-added milk bioconverted by L. plantarum SMFM2016-RK displayed both the growth inhibitory effect on periodontal pathogens and the α-glucosidase inhibitory activity; thus, it necessitates to evaluate the effects on the alleviation of periodontal diseases and glycemic control through future animal experiments. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
17. Evidence-Based Severity Assessment of Animal Models for Pancreatic Cancer.
- Author
-
Schreiber, Tim, Koopmann, Ingo, Brandstetter, Jakob, Talbot, Steven R., Goldstein, Lea, Hoffmann, Lisa, Schildt, Anna, Joksch, Markus, Krause, Bernd, Jaster, Robert, Palme, Rupert, Zechner, Dietmar, Vollmar, Brigitte, and Kumstel, Simone
- Subjects
PANCREATIC cancer ,NEST building ,PSYCHOLOGICAL distress ,ANIMAL experimentation ,INTRAVENOUS injections - Abstract
Animal models are crucial to preclinical oncological research and drug development. Animal experiments must be performed in accordance with the 3R principles of replacement and reduction, if possible, and refinement where these procedures remain crucial. In addition, European Union legislations demand a continuous refinement approach, as well as pro- and retrospective severity assessment. In this study, an objective databased severity assessment was performed in murine models for pancreatic cancer induced by orthotopic, subcutaneous, or intravenous injection of Panc02 cells. Parameters such as body weight change, distress score, perianal temperature, mouse grimace scale, burrowing, nesting behavior, and the concentration of corticosterone in plasma and its metabolites in feces were monitored during tumor progression. The most important parameters were combined into a score and mapped against a reference data set by the Relative Severity Assessment procedure (RELSA) to obtain the maximum achieved severity for each animal (RELSA
max ). This scoring revealed a significantly higher RELSAmax for the orthotopic model than for the subcutaneous and intravenous models. However, compared to animal models such as pancreatitis and bile duct ligation, the pancreatic cancer models are shown to be less severe. Data-based animal welfare assessment proved to be a valuable tool for comparing the severity of differently induced cancer models. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
18. The Non-Denatured Processing of Brasenia schreberi Mucilage—Characteristics of Hydrodynamic Properties and the Effect on In Vivo Functions.
- Author
-
Ai, Tingyang, Wan, Jiawei, Yu, Xiujuan, Liu, Jiao, Yin, Cong, Yang, Lindong, Liu, Hong, and Qin, Rui
- Subjects
MUCILAGE ,GUT microbiome ,COLITIS ,ANIMAL experimentation - Abstract
Food non-denatured processes, such as freeze-drying and grinding, are commonly applied to raw materials with good bioactive functions. Although the functional components are maintained, whether structural and physical changes impact the in vivo function is often ignored in practical situations. Brasenia schreberi mucilage (BSM) has a significant alleviation effect on DSS-induced colitis. This work focused on the influence of non-denatured manufacture on the colonic benefits of BSM-based products. First, three forms of products including fresh mucilage (FM), freeze-dried products (FS), and freeze-dried powder (FP) were prepared. Then, their in vitro physiochemical properties were compared, analyzing their influence on the gut inflammation degree, microbial composition, and SCFA production in mice. The results suggested that the water retention rate of FS and FP was decreased to 34.59 ± 3.85%, and 9.93 ± 1.76%. The viscosity of FM, FS, and FP was 20.14 Pa∙s, 4.92 Pa∙s, and 0.41 Pa∙s, respectively. The freeze-drying and grinding process also damaged the lamellar microstructure of BSM. Then, animal tests showed that colitis mice intervened with FM, FS, and FP had disease activity scores of 2.03, 3.95, and 4.62. Meanwhile, FM notably changed the gut microbial composition and significantly increased propionate and butyrate levels. It seemed that the distinct colitis alleviation efficacy of BSM-based products is attributed to different hydrodynamic properties in the gut. FM had relatively higher viscosity and correspondingly high nutritional density in the gut lumen, which stimulates Firmicutes growth and promotes butyrate production, and thereby exhibited the best efficiency on protecting from colitis. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
19. P18: Novel Anticancer Peptide from Induced Tumor-Suppressing Cells Targeting Breast Cancer and Bone Metastasis.
- Author
-
Cui, Changpeng, Huo, Qingji, Xiong, Xue, Na, Sungsoo, Mitsuda, Masaru, Minami, Kazumasa, Li, Baiyan, and Yokota, Hiroki
- Subjects
COMPUTER-assisted molecular modeling ,FLUORESCENCE polarization immunoassay ,RESEARCH funding ,BREAST tumors ,ANTINEOPLASTIC agents ,TRYPSIN ,ANTIMICROBIAL peptides ,TREATMENT effectiveness ,CANCER cell culture ,DESCRIPTIVE statistics ,CELL motility ,BONE metastasis ,MICE ,CELL lines ,ANIMAL experimentation ,WESTERN immunoblotting ,ONE-way analysis of variance ,CELL survival ,PHARMACODYNAMICS - Abstract
Simple Summary: This study focused on finding effective anticancer peptides (ACPs) from induced tumor-suppressing cells (iTSCs) to combat breast cancer metastasis to the skeletal system. This study identified P18 as the most potent ACP from a pool of candidates, derived from a protein called Arhgdia. P18 showed significant inhibitory effects on breast cancer cell viability, migration, and invasion, as well as interfering with certain cancer-promoting proteins and GTPase signaling. Importantly, P18 demonstrated enhanced effectiveness when combined with traditional chemotherapy drugs, without significantly affecting healthy cells. Moreover, it mitigated the bone loss associated with breast cancer spread to bones. This study suggests that P18, especially in its modified form (Ac-P18-NH2), could be a promising candidate for breast cancer treatment and preventing bone destruction by regulating specific cellular signaling pathways. Background: The skeletal system is a common site for metastasis from breast cancer. In our prior work, we developed induced tumor-suppressing cells (iTSCs) capable of secreting a set of tumor-suppressing proteins. In this study, we examined the possibility of identifying anticancer peptides (ACPs) from trypsin-digested protein fragments derived from iTSC proteomes. Methods: The efficacy of ACPs was examined using an MTT-based cell viability assay, a Scratch-based motility assay, an EdU-based proliferation assay, and a transwell invasion assay. To evaluate the mechanism of inhibitory action, a fluorescence resonance energy transfer (FRET)-based GTPase activity assay and a molecular docking analysis were conducted. The efficacy of ACPs was also tested using an ex vivo cancer tissue assay and a bone microenvironment assay. Results: Among the 12 ACP candidates, P18 (TDYMVGSYGPR) demonstrated the most effective anticancer activity. P18 was derived from Arhgdia, a Rho GDP dissociation inhibitor alpha, and exhibited inhibitory effects on the viability, migration, and invasion of breast cancer cells. It also hindered the GTPase activity of RhoA and Cdc42 and downregulated the expression of oncoproteins such as Snail and Src. The inhibitory impact of P18 was additive when it was combined with chemotherapeutic drugs such as Cisplatin and Taxol in both breast cancer cells and patient-derived tissues. P18 had no inhibitory effect on mesenchymal stem cells but suppressed the maturation of RANKL-stimulated osteoclasts and mitigated the bone loss associated with breast cancer. Furthermore, the P18 analog modified by N-terminal acetylation and C-terminal amidation (Ac-P18-NH2) exhibited stronger tumor-suppressor effects. Conclusions: This study introduced a unique methodology for selecting an effective ACP from the iTSC secretome. P18 holds promise for the treatment of breast cancer and the prevention of bone destruction by regulating GTPase signaling. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
20. The PCV3 Cap Virus-like Particle Vaccine with the Chimeric PCV2-Neutralizing Epitope Gene Is Effective in Mice.
- Author
-
Wu, Xingchen, Wang, Qikai, Lu, Wang, Wang, Ying, Han, Zehao, Liang, Libin, Gao, Shimin, Ma, Haili, and Luo, Xiaomao
- Subjects
VIRUS-like particles ,PORCINE reproductive & respiratory syndrome ,CIRCOVIRUS diseases ,ESCHERICHIA coli ,ANIMAL experimentation ,SWINE industry - Abstract
Simple Summary: Porcine circovirus type 3 (PCV3) infection can lead to clinical symptoms in weaned piglets similar to porcine circovirus type 2 (PCV2) infection. The increasing incidence of PCV2 and PCV3 infections, along with the prevalence of coinfections, has led to significant economic losses in the swine industry. Currently, there are no commercial vaccines available for PCV3, and PCV2 vaccines do not provide cross-protection against PCV3. This study optimized the cap3 sequence based on the codon preferences of E. coli and mammalian cells, resulting in the production of highly immunogenic PCV3 virus-like particles (VLPs). Animal experiments with various immunization strategies showed that pCap3-Cap2E VLPs significantly enhanced both humoral and cellular immune responses. This improvement not only reduced the PCV3 viral load in the lungs but also alleviated lung damage. These findings offer valuable insights into a potential strategy for the efficient development of a PCV3 vaccine. Porcine circovirus type 3 (PCV3) infection can cause symptoms similar to those of porcine circovirus type 2 (PCV2) infection, and coinfections with both PCV2 and PCV3 are observed in the swine industry. Consequently, developing chimeric vaccines is essential to prevent and control porcine circovirus infections. In this study, we used both E. coli and mammalian expression systems to express PCV3 Cap (Cap3) and a chimeric gene containing the PCV2-neutralizing epitope within the PCV3 Cap (Cap3-Cap2E), which were assembled into virus-like particle (VLP) vaccines. We found that Cap3 lacking nuclear localization signal (NLS) could not form VLPs, while Cap3 with a His-tag successfully assembled into VLPs. Additionally, the chimeric of PCV2-neutralizing epitopes did not interfere with the assembly process of VLPs. Various immunization approaches revealed that pCap3-Cap2E VLP vaccines were capable of activating high PCV3 Cap-specific antibody levels and effectively neutralizing both PCV3 and PCV2. Furthermore, pCap3-Cap2E VLPs demonstrated a potent ability to activate cellular immunity, protecting against PCV3 infection and preventing lung damage in mice. In conclusion, this study successfully developed a PCV3 Cap VLP vaccine incorporating chimeric PCV2-neutralizing epitope genes, providing new perspectives for PCV3 vaccine development. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
21. Understanding the Liver's Role in the Clearance of Aβ40.
- Author
-
Lockwood, Glen P., Hunt, Nicholas J., Kockx, Maaike, Kang, Sun Woo Sophie, Le Couteur, David G., and Cogger, Victoria C.
- Subjects
LIVER physiology ,IN vitro studies ,ALZHEIMER'S disease ,INDICATOR dilution ,FLUORESCENT antibody technique ,PEPTIDES ,RATS ,MICE ,LIVER cells ,ANIMAL experimentation ,AGING ,WESTERN immunoblotting ,BIOMARKERS - Abstract
The clearance of peripheral beta amyloid (Aβ) is a potential target for the treatment of Alzheimer's disease (AD). The liver has been implicated in the elimination of Aβ from the peripheral circulation. Here, the single-pass uptake of Aβ40 in perfused livers from young and old rats (6 to 10 rats per group) was investigated with the multiple indicator dilution technique. Aβ40 had volumes of distribution between those of the vascular marker Evans Blue and the extracellular marker sucrose. The hepatic extraction of Aβ40 was negligible, explained in part by the small permeability surface area products consistent with a high endothelial barrier to liver uptake. There were no substantial effects of age on any of these results. In vitro experiments with isolated hepatocytes and liver sinusoidal endothelial cells showed only very small amounts of Aβ uptake consistent with low intrinsic clearance. These results indicate that the hepatic clearance of Aβ is capacity-limited, explained by the low-permeability surface area products and hepatocyte uptake. However, this does not preclude an effect of aging in longer-term in vivo studies where age-related changes in liver blood flow and protein binding influence liver clearance. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
22. Expanding Understanding of Urban Rift Valley Fever Risk and Associated Vector Ecology at Slaughterhouses in Kisumu, Kenya.
- Author
-
Gerken, Keli Nicole, Owuor, Kevin Omondi, Ndenga, Bryson, Wambua, Sammy, Winter, Christabel, Chemutai, Salome, Omukuti, Rodney, Arabu, Daniel, Miring'u, Irene, Wilson, William C., Mutuku, Francis, Waggoner, Jesse J., Pinsky, Benjamin, Bosire, Carren, and LaBeaud, Angelle Desiree
- Subjects
RIFT Valley fever ,SLAUGHTERING ,URBAN ecology ,ANIMAL experimentation ,CULEX - Abstract
Rift Valley fever virus (RVFV) is an adaptable arbovirus that can be transmitted by a wide variety of arthropods. Widespread urban transmission of RVFV has not yet occurred, but peri-urban outbreaks of RVFV have recently been documented in East Africa. We previously reported low-level exposure in urban communities and highlighted the risk of introduction via live animal influx. We deployed a slaughtered animal testing framework in response to an early warning system at two urban slaughterhouses and tested animals entering the meat value chain for anti-RVFV IgG and IgM antibodies. We simultaneously trapped mosquitoes for RVFV and bloodmeal testing. Out of 923 animals tested, an 8.5% IgG seroprevalence was identified but no evidence of recent livestock exposure was detected. Mosquito species abundance varied greatly by slaughterhouse site, which explained 52% of the variance in blood meals. We captured many Culex spp., a known RVFV amplifying vector, at one of the sites (p < 0.001), and this species had the most diverse blood meals. No mosquito pools tested positive for RVFV antigen using a rapid VecTOR test. These results expand understanding of potential RVF urban disease ecology, and highlight that slaughterhouses are key locations for future surveillance, modelling, and monitoring efforts. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
23. Adipokines as Possible Players in Inflammatory Bowel Disease: Electrophysiological Evaluation of Their Role in Causing Functional Gastrointestinal Alterations in Murine Tissue.
- Author
-
Garella, Rachele, Palmieri, Francesco, and Squecco, Roberta
- Subjects
ADIPOKINES ,T-test (Statistics) ,SMOOTH muscle ,RESEARCH funding ,GASTROINTESTINAL motility ,APPETITE ,DESCRIPTIVE statistics ,INFLAMMATORY bowel diseases ,ADIPONECTIN ,GASTRIC fundus ,MICE ,PERMEABILITY ,DUMPING syndrome ,ANIMAL experimentation ,DATA analysis software ,RESISTIN ,ELECTROPHYSIOLOGY ,DISEASE progression ,ELECTRODES - Abstract
Inflammatory bowel disease (IBD) is a clinical condition of the gastrointestinal tract that has significant incidence in childhood. Major symptoms include abdominal pain, dyspepsia, delayed gastric emptying, anorexia, diarrhea and weight loss. IBD etiopathogenesis is multifactorial, with a proven involvement of cytokines. In this regard, cytokines like resistin and adiponectin produced by adipose tissue play a crucial role in inflammation. Particularly, resistin seems related to IBD severity and is considered a promising marker of disease occurrence and progression. Unraveling its mechanism of action and downstream effectors is mandatory when designing novel therapies. This preclinical study aims to further elucidate the action of resistin in causing functional gastrointestinal alterations, comparing it with the well-defined effect of adiponectin. To this end, we carried out electrophysiological analysis on murine gastric fundus. We found that resistin, similarly to adiponectin, increases smooth muscle cell (SMC) capacitance, indicative of cell surface remodeling, which is consistent with relaxation. However, contrary to adiponectin, resistin unalters membrane potential and inward Ca
2+ entry and scarcely affects outward current, suggesting its inefficacy in markedly modifying electrical phenomena on the SMC membrane. This outcome, supporting the role of resistin in gastrointestinal distention, as observed in IBD, rules out a strikingly direct effect on SMCs. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
24. Integrating Ataxia Evaluation into Tumor-Induced Hearing Loss Model to Comprehensively Study NF2-Related Schwannomatosis.
- Author
-
Lu, Simeng, Yin, Zhenzhen, Chen, Jie, Wu, Limeng, Sun, Yao, Gao, Xing, Huang, Peigen, Jordan, Justin T., Plotkin, Scott R., and Xu, Lei
- Subjects
THERAPEUTIC use of antineoplastic agents ,MOTOR ability ,ATAXIA ,RESEARCH funding ,NEUROFIBROMATOSIS 2 ,SENSORINEURAL hearing loss ,EARACHE ,DESCRIPTIVE statistics ,MICE ,TINNITUS ,ACOUSTIC neuroma ,QUALITY of life ,ANIMAL experimentation ,SCHWANNOMAS ,VESTIBULAR apparatus diseases ,HEARING disorders ,FACIAL paralysis ,BRAIN tumors ,MENTAL depression ,SYMPTOMS - Abstract
Simple Summary: The hallmark of NF2 is bilateral vestibular schwannomas, which progressively enlarge, leading to sensorineural hearing loss, tinnitus, facial weakness, and pain that translates to social impairment and clinical depression. To better understand disease progression and characterize treatment response, we developed a panel of five tests suitable for the mouse vestibular schwannoma model and investigated how tumor growth and treatment affect gait, coordination, and motor function. These methods, paired with hearing tests, enable a comprehensive evaluation of tumor-induced neurological deficits and facilitate the assessment of the effectiveness of novel therapeutics to improve NF2 treatments. NF2-related Schwannomatosis (NF2-SWN) is a disease that needs new solutions. The hallmark of NF2-SWN, a dominantly inherited neoplasia syndrome, is bilateral vestibular schwannomas (VSs), which progressively enlarge, leading to sensorineural hearing loss, tinnitus, facial weakness, and pain that translates to social impairment and clinical depression. Standard treatments for growing VSs include surgery and radiation therapy (RT); however, both carry the risk of further nerve damage that can result in deafness and facial palsy. The resultant suffering and debility, in combination with the paucity of therapeutic options, make the effective treatment of NF2-SWN a major unmet medical need. A better understanding of these mechanisms is essential to developing novel therapeutic targets to control tumor growth and improve patients' quality of life. Previously, we developed the first orthotopic cerebellopontine angle mouse model of VSs, which faithfully mimics tumor-induced hearing loss. In this model, we observed that mice exhibit symptoms of ataxia and vestibular dysfunction. Therefore, we further developed a panel of five tests suitable for the mouse VS model and investigated how tumor growth and treatment affect gait, coordination, and motor function. Using this panel of ataxia tests, we demonstrated that both ataxia and motor function deteriorated concomitantly with tumor progression. We further demonstrated that (i) treatment with anti-VEGF resulted in tumor size reduction, mitigated ataxia, and improved rotarod performance; (ii) treatment with crizotinib stabilized tumor growth and led to improvements in both ataxia and rotarod performance; and (iii) treatment with losartan did not impact tumor growth nor ameliorate ataxia or motor function. Our studies demonstrated that these methods, paired with hearing tests, enable a comprehensive evaluation of tumor-induced neurological deficits and facilitate the assessment of the effectiveness of novel therapeutics to improve NF2 treatments. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
25. Sex Matters–Insights from Testing Drug Efficacy in an Animal Model of Pancreatic Cancer.
- Author
-
Schulz, Benjamin, Leitner, Emily, Schreiber, Tim, Lindner, Tobias, Schwarz, Rico, Aboutara, Nadine, Ma, Yixuan, Escobar, Hugo Murua, Palme, Rupert, Hinz, Burkhard, Vollmar, Brigitte, and Zechner, Dietmar
- Subjects
BIOLOGICAL models ,RESEARCH funding ,SEX distribution ,IN vivo studies ,CELLULAR signal transduction ,PANCREATIC tumors ,MICE ,SMALL molecules ,DRUG efficacy ,ANIMAL experimentation ,DUCTAL carcinoma ,LUNG tumors ,TRANSFERASES - Abstract
Simple Summary: Pancreatic ductal adenocarcinoma continues to be one of the deadliest cancers worldwide. Preclinical studies involving animals rarely include sex as a major biological variable in testing the efficacy of new drugs. In an animal model of pancreatic cancer, we analyzed the impact of sex on the pathological features of the disease and on an experimental small molecule-based therapy tested in vivo for the first time. While the therapy shows potential, the obtained results are confounded by sex-specific effects. This study, therefore, highlights the importance of sex-inclusive research while simultaneously providing a basis for further studies of the therapy tested. Preclinical studies rarely test the efficacy of therapies in both sexes. The field of oncology is no exception in this regard. In a model of syngeneic, orthotopic, metastasized pancreatic ductal adenocarcinoma we evaluated the impact of sex on pathological features of this disease as well as on the efficacy and possible adverse side effects of a novel, small molecule-based therapy inhibiting KRAS:SOS1, MEK1/2 and PI3K signaling in male and female C57BL/6J mice. Male mice had less tumor infiltration of CD8-positive cells, developed bigger tumors, had more lung metastasis and a lower probability of survival compared to female mice. These more severe pathological features in male animals were accompanied by higher distress at the end of the experiment. The evaluated inhibitors BI-3406, trametinib and BKM120 showed synergistic effects in vitro. This combinatorial therapy reduced tumor weight more efficiently in male animals, although the drug concentrations were similar in the tumors of both sexes. These results underline the importance of sex-specific preclinical research and at the same time provide a solid basis for future studies with the tested compounds. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
26. Osteosarcoma-Induced Pain Is Mediated by Glial Cell Activation in the Spinal Dorsal Horn, but Not Capsaicin-Sensitive Nociceptive Neurons: A Complex Functional and Morphological Characterization in Mice.
- Author
-
Bencze, Noémi, Scheich, Bálint, Szőke, Éva, Wilhelm, Imola, Körmöndi, Sándor, Botz, Bálint, and Helyes, Zsuzsanna
- Subjects
OSTEOSARCOMA ,BIOLOGICAL models ,CARRIER proteins ,RESEARCH funding ,NEUROGLIA ,NEURONS ,NOCICEPTIVE pain ,BONE tumors ,PAIN threshold ,NEUROINFLAMMATION ,MICE ,IMMUNOHISTOCHEMISTRY ,HYPERALGESIA ,PAIN ,ANIMAL experimentation ,PAIN management ,CAPSAICIN ,SPINAL cord ,CELL receptors - Abstract
Simple Summary: Bone cancer-related chronic pain is mediated by central sensitization involving neuroinflammation via glial cell activation in the spinal dorsal horn, but not the capsaicin-sensitive sensory neuronal system. Bone cancer and its related chronic pain are huge clinical problems since the available drugs are often ineffective or cannot be used long term due to a broad range of side effects. The mechanisms, mediators and targets need to be identified to determine potential novel therapies. Here, we characterize a mouse bone cancer model induced by intratibial injection of K7M2 osteosarcoma cells using an integrative approach and investigate the role of capsaicin-sensitive peptidergic sensory nerves. The mechanical pain threshold was assessed by dynamic plantar aesthesiometry, limb loading by dynamic weight bearing, spontaneous pain-related behaviors via observation, knee diameter with a digital caliper, and structural changes by micro-CT and glia cell activation by immunohistochemistry in BALB/c mice of both sexes. Capsaicin-sensitive peptidergic sensory neurons were defunctionalized by systemic pretreatment with a high dose of the transient receptor potential vanilloid 1 (TRPV1) agonist resiniferatoxin (RTX). During the 14- and 28-day experiments, weight bearing on the affected limb and the paw mechanonociceptive thresholds significantly decreased, demonstrating secondary mechanical hyperalgesia. Signs of spontaneous pain and osteoplastic bone remodeling were detected both in male and female mice without any sex differences. Microglia activation was shown by the increased ionized calcium-binding adapter molecule 1 (Iba1) immunopositivity on day 14 and astrocyte activation by the enhanced glial fibrillary acidic protein (GFAP)-positive cell density on day 28 in the ipsilateral spinal dorsal horn. Interestingly, defunctionalization of the capsaicin-sensitive afferents representing approximately 2/3 of the nociceptive fibers did not alter any functional parameters. Here, we provide the first complex functional and morphological characterization of the K7M2 mouse osteosarcoma model. Bone-cancer-related chronic pain and hyperalgesia are likely to be mediated by central sensitization involving neuroinflammation via glial cell activation in the spinal dorsal horn, but not the capsaicin-sensitive sensory neuronal system. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
27. Evolution Characterization and Pathogenicity of an NADC34-like PRRSV Isolated from Inner Mongolia, China.
- Author
-
Zhao, Hong-Zhe, Liu, Chun-Yu, Meng, Hai, Sun, Cheng-Long, Yang, Hong-Wen, Wang, Hao, Zou, Jian, Li, Peng, Han, Feng-Ye, Qi, Gen, Zhang, Yang, Lin, Bing-Bing, Liu, Chuang, Chen, Meng-Meng, Zhang, Pan-Ling, Chen, Xiao-Dong, Zhang, Yi-Di, Song, Qian-Jin, Wen, Yong-Jun, and Wang, Feng-Xue
- Subjects
PORCINE reproductive & respiratory syndrome ,LUNGS ,PATHOLOGICAL physiology ,PIGLETS ,ANIMAL experimentation - Abstract
Porcine reproductive and respiratory syndrome virus (PRRSV) is a pathogen that causes severe abortions in sows and high piglet mortality, resulting in huge economic losses to the pig industry worldwide. The emerging and novel PRRSV isolates are clinically and biologically important, as there are likely recombination and pathogenic differences among PRRSV genomes. Furthermore, the NADC34-like strain has become a major epidemic strain in some parts of China, but the characterization and pathogenicity of the latest strain in Inner Mongolia have not been reported in detail. In this study, an NADC34-like strain (CHNMGKL1-2304) from Tongliao City, Inner Mongolia was successfully isolated and characterized, and confirmed the pathogenicity in pigs. The phylogenetic tree showed that this strain belonged to sublineage 1.5 and had high homology with the strain JS2021NADC34. There is no recombination between CHNMGKL1-2304 and any other domestic strains. Animal experiments show that the CHNMGKL1-2304 strain is moderately virulent to piglets, which show persistent fever, weight loss and high morbidity but no mortality. The presence of PRRSV nucleic acids was detected in both blood, tissues, nasal and fecal swabs. In addition, obvious pathological changes and positive signals were observed in lung, lymph node, liver and spleen tissues when subjected to hematoxylin–eosin (HE) staining and immunohistochemistry (IHC). This report can provide a basis for epidemiological investigations and subsequent studies of PRRSV. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
28. Opaganib Downregulates N-Myc Expression and Suppresses In Vitro and In Vivo Growth of Neuroblastoma Cells.
- Author
-
Maines, Lynn W., Keller, Staci N., Smith, Ryan A., Schrecengost, Randy S., and Smith, Charles D.
- Subjects
IN vitro studies ,PROTEINS ,IRINOTECAN ,COMBINATION drug therapy ,RESEARCH funding ,PATIENT safety ,CELL proliferation ,TEMOZOLOMIDE ,IN vivo studies ,XENOGRAFTS ,IMMUNODIAGNOSIS ,ORAL drug administration ,CELLULAR signal transduction ,GENE expression ,CELL culture ,SPHINGOLIPIDS ,MICE ,IMMUNE checkpoint inhibitors ,SPHINGOSINE-1-phosphate ,ANIMAL experimentation ,COMPARATIVE studies ,NEUROBLASTOMA ,CELL surface antigens ,DRUG tolerance ,CHEMICAL inhibitors - Abstract
Simple Summary: Neuroblastoma is the most common cancer in infants and the most common solid tumor outside the brain in children, and responds poorly to current therapies. The sphingolipid-modifying drug opaganib, which has anticancer and anti-inflammatory activity in many preclinical models, was tested for inhibition of neuroblastoma cell proliferation in cell culture and in tumors growing in mice. Opaganib inhibited cell proliferation regardless of the MYCN gene status of the neuroblastoma cells. Treatment of tumor-bearing mice with opaganib in combination with the established drugs irinotecan and temozolomide reduced tumor growth and increased survival compared with irinotecan plus temozolomide alone. Because opaganib has already demonstrated safety in patients with cancer, this new drug may provide improved therapy for neuroblastoma patients. Neuroblastoma (NB), the most common cancer in infants and the most common solid tumor outside the brain in children, grows aggressively and responds poorly to current therapies. We have identified a new drug (opaganib, also known as ABC294640) that modulates sphingolipid metabolism by inhibiting the synthesis of sphingosine 1-phosphate (S1P) by sphingosine kinase-2 and elevating dihydroceramides by inhibition of dihydroceramide desaturase. The present studies sought to determine the potential therapeutic activity of opaganib in cell culture and xenograft models of NB. Cytotoxicity assays demonstrated that NB cells, including cells with amplified MYCN, are effectively killed by opaganib concentrations well below those that accumulate in tumors in vivo. Opaganib was shown to cause dose-dependent decreases in S1P and hexosylceramide levels in Neuro-2a cells, while concurrently elevating levels of dihydroceramides. As with other tumor cells, opaganib reduced c-Myc and Mcl-1 protein levels in Neuro-2a cells, and also reduced the expression of the N-Myc protein. The in vivo growth of xenografts of human SK-N-(BE)2 cells with amplified MYCN was suppressed by oral administration of opaganib at doses that are well tolerated in mice. Combining opaganib with temozolomide plus irinotecan, considered the backbone for therapy of relapsed or refractory NB, resulted in increased antitumor activity in vivo compared with temozolomide plus irinotecan or opaganib alone. Mice did not lose additional weight when opaganib was combined with temozolomide plus irinotecan, indicating that the combination is well tolerated. Opaganib has additive antitumor activity toward Neuro-2a tumors when combined with the checkpoint inhibitor anti-CTLA-4 antibody; however, the combination of opaganib with anti-PD-1 or anti-PD-L1 antibodies did not provide increased antitumor activity over that seen with opaganib alone. Overall, the data demonstrate that opaganib modulates sphingolipid metabolism and intracellular signaling in NB cells and inhibits NB tumor growth alone and in combination with other anticancer drugs. Amplified MYCN does not confer resistance to opaganib, and, in fact, the drug attenuates the expression of both c-Myc and N-Myc. The safety of opaganib has been established in clinical trials with adults with advanced cancer or severe COVID-19, and so opaganib has excellent potential for treating patients with NB, particularly in combination with temozolomide and irinotecan or anti-CTLA-4 antibody. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
29. Activation of the Anaphase Promoting Complex Restores Impaired Mitotic Progression and Chemosensitivity in Multiple Drug-Resistant Human Breast Cancer.
- Author
-
Lubachowski, Mathew, VanGenderen, Cordell, Valentine, Sarah, Belak, Zach, Davies, Gerald Floyd, Arnason, Terra Gayle, and Harkness, Troy Anthony Alan
- Subjects
IN vitro studies ,FLOW cytometry ,RESEARCH funding ,T-test (Statistics) ,BREAST tumors ,CELL physiology ,APOPTOSIS ,ANIMALS ,KRUSKAL-Wallis Test ,CELL division ,CELL lines ,MICE ,CELL culture ,ANIMAL experimentation ,DNA damage ,WESTERN immunoblotting ,DRUG resistance ,PHENOTYPES - Abstract
Simple Summary: Patients with treatable cancers all too often return to the clinic with untreatable tumors, requiring highly toxic secondary treatments or palliative care. In this study, we aim to determine whether activation of the Anaphase Promoting Complex (APC) in multiple-drug-resistant (MDR) breast cancer cells will re-sensitize them to chemotherapeutic agents. The APC is required for the targeted degradation of proteins that inhibit passage through mitosis. We found that APC activity is indeed impaired in MDR cells and that chemical activation of the APC increased the sensitivity of these cells to doxorubicin. Mitotic progression was slowed in MDR cells, compared to matched parental drug sensitive cells, with an accumulation of APC substrate proteins. APC activation in nocodazole-arrested cells resulted in increased passage through mitosis with lower APC substrate levels. Mice growing a patient-derived xenografted (PDX) tumor were treated with increasing doses of a chemical APC activator, resulting in a dose-dependent reduction in tumor size. Taken together, our data show that APC activity is reduced in MDR cells, with APC activation resulting in a species- and cancer-type-independent reversal of the MDR phenotype. The development of multiple-drug-resistant (MDR) cancer all too often signals the need for toxic alternative therapy or palliative care. Our recent in vivo and in vitro studies using canine MDR lymphoma cancer cells demonstrate that the Anaphase Promoting Complex (APC) is impaired in MDR cells compared to normal canine control and drug-sensitive cancer cells. Here, we sought to establish whether this phenomena is a generalizable mechanism independent of species, malignancy type, or chemotherapy regime. To test the association of blunted APC activity with MDR cancer behavior, we used matched parental and MDR MCF7 human breast cancer cells, and a patient-derived xenograft (PDX) model of human triple-negative breast cancer. We show that APC activating mechanisms, such as APC subunit 1 (APC1) phosphorylation and CDC27/CDC20 protein associations, are reduced in MCF7 MDR cells when compared to chemo-sensitive matched cell lines. Consistent with impaired APC function in MDR cells, APC substrate proteins failed to be effectively degraded. Similar to our previous observations in canine MDR lymphoma cells, chemical activation of the APC using Mad2 Inhibitor-1 (M2I-1) in MCF7 MDR cells enhanced APC substrate degradation and resensitized MDR cells in vitro to the cytotoxic effects of the alkylating chemotherapeutic agent, doxorubicin (DOX). Using cell cycle arrest/release experiments, we show that mitosis is delayed in MDR cells with elevated substrate levels. When pretreated with M2I-1, MDR cells progress through mitosis at a faster rate that coincides with reduced levels of APC substrates. In our PDX model, mice growing a clinically MDR human triple-negative breast cancer tumor show significantly reduced tumor growth when treated with M2I-1, with evidence of increased DNA damage and apoptosis. Thus, our results strongly support the hypothesis that APC impairment is a driver of aggressive tumor development and that targeting the APC for activation has the potential for meaningful clinical benefits in treating recurrent cases of MDR malignancy. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
30. Proteins Involved in Focal Cell Adhesion and Podosome Formation Are Differentially Expressed during Colorectal Tumorigenesis in AOM-Treated Rats.
- Author
-
Swain, Ian X. and Kresak, Adam M.
- Subjects
ABERRANT crypt foci ,PROTEINS ,ADENOCARCINOMA ,NEOPLASTIC cell transformation ,RESEARCH funding ,CELL adhesion molecules ,COLORECTAL cancer ,CELLULAR signal transduction ,TUMOR markers ,GENE expression ,RATS ,ANIMAL experimentation ,WESTERN immunoblotting ,ENDOTHELIAL cells - Abstract
Simple Summary: Colon cancer first involves the development of small clusters of abnormal cells. The purpose of our study was to determine changes in protein expression in such abnormal cells as they grow after being triggered to form due to exposure to a carcinogen using rats as a model. We were able to identify a number of proteins that were very differently expressed when comparing early abnormal cells to more mature cell clusters. Many of these proteins are involved in regulating cell-to-cell interactions and cell shape, among other activities in the cells. Our findings help us better understand the biology of intestinal cancer and may be useful in developing biomarkers for detecting colon cancer. This information also helps us lay a foundation for future studies aimed at better understanding the how intestinal cancers develop and grow under different intestinal conditions. Colorectal tumorigenesis involves the development of aberrant crypt foci (ACF) or preneoplastic lesions, representing the earliest morphological lesion visible in colon cancer. The purpose of this study was to determine changes in protein expression in carcinogen-induced ACF as they mature and transform into adenomas. Protein expression profiles of azoxymethane (AOM)-induced F344 rat colon ACF and adenomas were compared at four time points, 4 (control), 8, 16, and 24 weeks post AOM administration (n = 9/group), with time points correlating with induction and transformation events. At each time point, micro-dissected ACF and/or adenoma tissues were analyzed across multiple quantitative two-dimensional (2D-DIGE) gels using a Cy-dye labeling technique and a pooled internal standard to quantify expression changes with statistical confidence. Western blot and subsequent network pathway mapping were used to confirm and elucidate differentially expressed (p ≤ 0.05) proteins, including changes in vinculin (Vcl; p = 0.007), scinderin (Scin; p = 0.02), and profilin (Pfn1; p = 0.01), By determining protein expression changes in ACF as they mature and transform into adenomas, a "baseline" of altered regulatory proteins associated with adenocarcinoma development in this model has been elucidated. These data will enable future studies aimed at biomarker identification and understanding the molecular biology of intestinal tumorigenesis and adenocarcinoma maturation under varying intestinal conditions. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
31. Experimental Models to Study Immune Dysfunction in the Pathogenesis of Parkinson's Disease.
- Author
-
Saponjic, Jasna, Mejías, Rebeca, Nikolovski, Neda, Dragic, Milorad, Canak, Asuman, Papoutsopoulou, Stamatia, Gürsoy-Özdemir, Yasemin, Fladmark, Kari E., Ntavaroukas, Panagiotis, Bayar Muluk, Nuray, Zeljkovic Jovanovic, Milica, Fontán-Lozano, Ángela, Comi, Cristoforo, and Marino, Franca
- Subjects
PARKINSON'S disease ,PATHOGENESIS ,CENTRAL nervous system ,DISEASE progression ,ANIMAL experimentation - Abstract
Parkinson's disease (PD) is a chronic, age-related, progressive multisystem disease associated with neuroinflammation and immune dysfunction. This review discusses the methodological approaches used to study the changes in central and peripheral immunity in PD, the advantages and limitations of the techniques, and their applicability to humans. Although a single animal model cannot replicate all pathological features of the human disease, neuroinflammation is present in most animal models of PD and plays a critical role in understanding the involvement of the immune system (IS) in the pathogenesis of PD. The IS and its interactions with different cell types in the central nervous system (CNS) play an important role in the pathogenesis of PD. Even though culture models do not fully reflect the complexity of disease progression, they are limited in their ability to mimic long-term effects and need validation through in vivo studies. They are an indispensable tool for understanding the interplay between the IS and the pathogenesis of this disease. Understanding the immune-mediated mechanisms may lead to potential therapeutic targets for the treatment of PD. We believe that the development of methodological guidelines for experiments with animal models and PD patients is crucial to ensure the validity and consistency of the results. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
32. Storytelling of Myocarditis.
- Author
-
Thiene, Gaetano
- Subjects
MYOCARDITIS ,CARDIAC magnetic resonance imaging ,EXTRACORPOREAL membrane oxygenation ,COXSACKIEVIRUSES ,ANIMAL experimentation - Abstract
In 1900, Fiedler first reported autopsy cases with peculiar inflammation of the myocardium, which he named interstitial myocarditis. He postulated an isolated cardiac inflammation of the myocardium in the absence of multiorgan involvement and with a poor prognosis due to invisible microorganisms, which years later would have been identified as viruses. The revision of original histologic sections by Schmorl showed cases with lymphocytes and others with giant-cell inflammatory histotypes. The in vivo diagnosis of myocarditis became possible thanks to right cardiac catheterization with endomyocardial biopsy (EMB). The gold standard for diagnosis was achieved with the employment of immunohistochemistry and molecular investigation by Polymerase Chain Reaction (PCR), which allows for the detection of viruses as causal agents. Both RNA and DNA were revealed to be cardiotropic, with a common receptor (CAR). A protease, coded by coxsackie virus, disrupts the cytoskeleton and accounts for cell death. Unfortunately, vaccination, despite having been revealed to be effective in animal experiments, has not yet entered the clinical field for prevention. Cardiac Magnetic Resonance turned out to be a revolutionary tool for in vivo diagnosis through the detection of edema (inflammatory exudate). Myocarditis may be fulminant in terms of clinical presentation but not necessarily fatal. The application of ExtraCorporeal Membrane Oxygenation (ECMO) allows for relieving the overloaded native heart. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
33. Magnesium Is a Vital Ion in the Body—It Is Time to Consider Its Supplementation on a Routine Basis.
- Author
-
Pethő, Ákos Géza, Fülöp, Tibor, Orosz, Petronella, and Tapolyai, Mihály
- Subjects
MAGNESIUM ,DISEASE risk factors ,ARTERIAL calcification ,ANIMAL experimentation ,DIETARY supplements - Abstract
The importance of maintaining proper magnesium intake and total body magnesium content in preserving human health remains underappreciated among medical professionals and laymen. This review aimed to show the importance of hypomagnesemia as a modifiable risk factor for developing disease processes. We searched the PubMed database and Google Scholar using the keywords 'magnesium', 'diabetes', 'cardiovascular disease', 'respiratory disease', 'immune system', 'inflammation', 'autoimmune disease', 'neurology', 'psychiatry', 'cognitive function', 'cancer', and 'vascular calcification'. In multiple contexts of the search terms, all reviews, animal experiments, and human observational data indicated that magnesium deficiency can lead to or contribute to developing many disease states. The conclusions of several in-depth reviews support our working hypothesis that magnesium and its supplementation are often undervalued and underutilized. Although much research has confirmed the importance of proper magnesium supply and tissue levels, simple and inexpensive magnesium supplementation has not yet been sufficiently recognized or promoted. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
34. Long-Term Oral Tamoxifen Administration Decreases Brain-Derived Neurotrophic Factor in the Hippocampus of Female Long-Evans Rats.
- Author
-
Been, Laura E., Halliday, Amanda R., Blossom, Sarah M., Bien, Elena M., Bernhard, Anya G., Roth, Grayson E., Domenech Rosario, Karina I., Pollock, Karlie B., Abramenko, Petra E., Behbehani, Leily M., Pascal, Gabriel J., and Kelly, Mary Ellen
- Subjects
BIOLOGICAL models ,LIQUID chromatography-mass spectrometry ,RESEARCH funding ,BLOOD collection ,ORAL drug administration ,TAMOXIFEN ,RATS ,IMMUNOHISTOCHEMISTRY ,BRAIN-derived neurotrophic factor ,ANIMAL experimentation ,HIPPOCAMPUS (Brain) - Abstract
Simple Summary: Tamoxifen is prescribed to premenopausal patients with estrogen-receptor-positive breast cancers for a period of 5–10 years after cancer diagnosis. This prolonged treatment regimen, though effective at preventing cancer recurrence, is often associated with unwanted cognitive and affective symptoms. To understand the clinical side effects of tamoxifen, animal studies investigating the effect of tamoxifen on the brain must model the chronic nature of tamoxifen therapy. This study describes a novel method of tamoxifen administration in female rats conducive to long-term administration of tamoxifen. Blood samples from treated rats showed levels of tamoxifen similar to levels in humans. Brain samples revealed tamoxifen-induced changes of a neurotrophic factor in the hippocampus, a structure critical to cognitive and affective processing. This study, therefore, suggests a potential mechanism that may underlie the cognitive side effects reported in patients. Tamoxifen, a selective estrogen receptor modulator (SERM), is commonly used as an adjuvant drug therapy for estrogen-receptor-positive breast cancers. Though effective at reducing the rate of cancer recurrence, patients often report unwanted cognitive and affective side effects. Despite this, the impacts of chronic tamoxifen exposure on the brain are poorly understood, and rodent models of tamoxifen exposure do not replicate the chronic oral administration seen in patients. We, therefore, used long-term ad lib consumption of medicated food pellets to model chronic tamoxifen exposure in a clinically relevant way. Adult female Long-Evans Hooded rats consumed tamoxifen-medicated food pellets for approximately 12 weeks, while control animals received standard chow. At the conclusion of the experiment, blood and brain samples were collected for analyses. Blood tamoxifen levels were measured using a novel ultra-performance liquid chromatography–tandem mass spectrometry assay, which found that this administration paradigm produced serum levels of tamoxifen similar to those in human patients. In the brain, brain-derived neurotrophic factor (BDNF) was visualized in the hippocampus using immunohistochemistry. Chronic oral tamoxifen treatment resulted in a decrease in BDNF expression across several regions of the hippocampus. These findings provide a novel method of modeling and measuring chronic oral tamoxifen exposure and suggest a putative mechanism by which tamoxifen may cause cognitive and behavioral changes reported by patients. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
35. Animal Research Regulation: Improving Decision-Making and Adopting a Transparent System to Address Concerns around Approval Rate of Experiments.
- Author
-
Azilagbetor, David Mawufemor, Shaw, David, and Elger, Bernice Simone
- Subjects
LABORATORY animals ,PUBLIC opinion ,ATTITUDES toward the environment ,SOCIAL attitudes ,ANIMAL experimentation ,ANIMAL welfare - Abstract
Simple Summary: Scientific experiments that use animals are strictly regulated, mainly as a result of public concerns about animal testing. Examination of the regulation of animal experiments shows that most of the experiments submitted for authorization are approved. But what could be the reasons for this high approval? Since public understanding is needed in order to reach a common agreement that will shape animal research policy, it is important to have a transparent discussion about what leads to the frequent approval of animal experiments. The aim of this article is to discuss why frequent approval of experiments is a problem: the regulatory process has been put in place to authorize only acceptable experiments. Even though some experiments, such as cosmetic testing on animals, are prohibited in many countries irrespective of harm–benefit ratios, most types of animal experiments are approved. Further, we explain some possible genuine reasons for frequent approval, but at the same time, we also discuss how the regulatory process could be leading to the approval of unacceptable studies. To ensure transparency, gain public trust, and improve the understanding of the regulation of animal testing, we propose that regulators publish their experiment evaluations on an accessible online platform, explaining how they reached their decisions. The use of nonhuman animals in biomedical research is regulated under stringent conditions, not only in response to societal attitudes towards animal experimentation but also because ethical responsibility in scientific research requires researchers and veterinarians to be more invested and aim to improve the welfare of animals used for experiments. Analyses of animal research oversight reveal the frequent approval of experiments, and the approval of some experiments has raised and continues to raise public concerns. Societal compliance is required for a consensus-based approach to animal research policy, prompting the need to have transparent discussions about oversight and the frequency of approvals. We discuss how frequent approval may be perceived and why it seems problematic from a societal perspective: the regulatory process exists to approve only legitimate experiments. Although some experiments remain unacceptable irrespective of their harm–benefit ratios, almost all experiments are approved. We explain some possible legitimate reasons for frequent approval and how the review process could be leading to the approval of illegitimate studies. To ensure transparency and improve public trust and understanding of oversight, we propose the adoption of a platform to inform society about how unethical experiments are screened out. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
36. Exercise Suppresses Head and Neck Squamous Cell Carcinoma Growth via Oncostatin M.
- Author
-
Yoshimura, Takuya, Hirano, Yuka, Hamada, Taiji, Yokoyama, Seiya, Suzuki, Hajime, Takayama, Hirotaka, Migita, Hirono, Ishida, Takayuki, Nakamura, Yasunori, Ohsawa, Masahiro, Asakawa, Akihiro, Ishihata, Kiyohide, and Tanimoto, Akihide
- Subjects
SQUAMOUS cell carcinoma ,SURVIVAL ,MOUTH tumors ,RESEARCH funding ,EXERCISE therapy ,HEAD & neck cancer ,ENZYME-linked immunosorbent assay ,CELL proliferation ,METASTASIS ,ANIMAL experimentation ,CYTOKINES ,CELL survival ,DISEASE progression ,MYOKINES ,SARCOPENIA - Abstract
Simple Summary: In recent years, much research has focused on how exercise improves cancer prognosis and how the protein myokine, which is associated with exercise, inhibits cancer progression. However, there have been few detailed studies on oral cancer. This study is the first to show that exercise inhibits the progression of oral cancer. While other studies have shown that exercise suppresses growth, decreases the tumor formation rate, or improves survival in patients with other organ cancers, this study showed that exercise inhibits tumor formation and growth in oral cancer and prolongs survival. Furthermore, while many studies have demonstrated an indirect immune-mediated mechanism by which exercise suppresses cancer, this study suggests the additional possibility that myokines released by exercise directly affect oral cancer cells. This study has the above novelties. Major advances have been made in cancer treatment, but the prognosis for elderly cancer patients with sarcopenia and frailty remains poor. Myokines, which are thought to exert preventive effects against sarcopenia, have been reported to be associated with the prognosis of various cancers, but their effect on head and neck squamous cell carcinoma (HNSCC) is unknown. The aim of this study was to clarify the influence of exercise on the control of HNSCC and to examine the underlying mechanism involved. Mice were injected with HSC-3-M3 cells, a human cell line of highly metastatic and poorly differentiated tongue cancer, at the beginning of the study. Just prior to transplantation, blood was collected from the mice, and the levels of myokines were measured by ELISA. Oncostatin M (OSM), a selected myokine, was added to HSC-3-M3 cells, after which the cell proliferation ability, cell cycle, and protein expression were analyzed in vitro. Tumor cell viability was lower (control: 100%, exercise: 75%), tumors were smaller (control: 26.2 mm
3 , exercise: 6.4 mm3 ), and survival was longer in the exercise group than in the control group in vivo. OSM inhibited HSC-3-M3 cell proliferation in a concentration-dependent manner in vitro. The addition of OSM increased the proportion of cells in the G0/G1 phase, decreased the proportion of cells in the G2/M phase, and increased the expression of the CDK inhibitors p21 and p27. These results indicate that exercise may directly inhibit the proliferation of HNSCC cell lines via OSM. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
37. TIM3 Checkpoint Inhibition Fails to Prolong Survival in Ovarian Cancer-Bearing Mice.
- Author
-
Berckmans, Yani, Vankerckhoven, Ann, Caro, Aarushi Audhut, Kempeneers, Julie, Ceusters, Jolien, Thirion, Gitte, Vandenbrande, Katja, Vergote, Ignace, Laoui, Damya, and Coosemans, An
- Subjects
RESEARCH funding ,ANTINEOPLASTIC agents ,OVARIAN tumors ,TREATMENT effectiveness ,IMMUNE checkpoint inhibitors ,MICE ,CANCER chemotherapy ,ANIMAL experimentation - Abstract
Simple Summary: Ovarian cancer still ranks as the deadliest gynecological malignancy worldwide. In this research, we aimed to evaluate the potential of anti-TIM3, a relatively novel checkpoint inhibitor, for the treatment of ovarian cancer. Our preclinical studies showed no improvement of survival in ovarian cancer-bearing mice after anti-TIM3 treatment. Additionally, we found no significant immunological changes induced by this therapy in treated mice. However, changing the order of combination treatment with anti-TIM3 and chemotherapy influenced the outcome in mice. Further preclinical studies are required to find and optimize ovarian cancer treatment approaches. Immune checkpoint inhibitor (ICI) therapy has proven revolutionary in the treatment of some cancers. However, ovarian cancer remains unresponsive to current leading ICIs, such as anti-PD1 or anti-PD-L1. In this article, we explored the potential of an upcoming checkpoint molecule, T-cell immunoglobulin and mucin domain 3 (TIM3), for the treatment of ovarian cancer using a syngeneic orthotopic mouse model (ID8-fLuc). Besides therapeutic efficacy, we focused on exploring immune changes in tumor tissue and peritoneal fluid. Our results showed no improvement in survival in ovarian cancer-bearing mice after anti-TIM3 treatment when used as monotherapy nor when combined with anti-PD1 or standard-of-care chemotherapy (carboplatin/paclitaxel). This was reflected in the unaltered immune infiltration in treated mice compared to control mice. Altering the order of drug administration within the combination treatment altered the survival results, but did not result in a survival benefit over chemotherapy alone. These findings highlight the need for further preclinical studies to find beneficial treatment schemes and combination therapies for ovarian cancer. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
38. Mandibular Endochondral Growth Is Specifically Augmented by Nutritional Supplementation with Myo-Inositol Even in Rabbits.
- Author
-
Shimoyama, Miho, Kanzaki, Hiroyuki, Tohyama, Syunnosuke, Ida, Tomomi, Ishikawa, Misao, Katsumata, Yuta, Arai, Chihiro, Wada, Satoshi, Manase, Shugo, and Tomonari, Hiroshi
- Subjects
RABBITS ,DIETARY supplements ,ORTHODONTIC appliances ,ANIMAL experimentation ,THREE-dimensional imaging - Abstract
Mandibular retrognathism occurs by insufficient mandibular growth and causes several issues, such as respiratory difficulty and diminished masticatory function. At present, functional orthodontic appliances are used for stimulating mandibular growth in pediatric cases. However, the effectiveness of functional appliances is not always stable in daily practices. A more effective, reliable, and safer therapeutic method for mandibular growth promotion would be helpful for growing mandibular retrognathism patients. As we previously discovered that nutritional supplementation of myo-inositol in growing mice specifically increases mandibular endochondral growth, we performed preclinical animal experiments in rabbits in this study. Briefly, six-week-old male Japanese white rabbits were fed with or without myo-inositol supplementation in laboratory chow until 25 weeks old, and 3D image analysis using micro CT data and histological examinations was done. Myo-inositol had no systemic effect, such as femur length, though myo-inositol specifically augmented the mandibular growth. Myo-inositol increased the thickness of mandibular condylar cartilage. We discovered that the nutritional supplementation of myo-inositol during the growth period specifically augmented mandibular growth without any systemic influence, even in rabbits. Our results suggest the possibility of clinical use of myo-inositol for augmentation of the mandibular growth in growing mandibular retrognathism patients in the future. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
39. Longitudinal Muscle Biopsies Reveal Inter- and Intra-Subject Variability in Cancer Cachexia: Paving the Way for Biopsy-Guided Tailored Treatment.
- Author
-
Filis, Panagiotis, Tzavellas, Nikolaos P., Stagikas, Dimitrios, Zachariou, Christianna, Lekkas, Panagiotis, Kosmas, Dimitrios, Dounousi, Evangelia, Sarmas, Ioannis, Ntzani, Evangelia, Mauri, Davide, Korompilias, Anastasios, Simos, Yannis V., Tsamis, Konstantinos I., and Peschos, Dimitrios
- Subjects
CACHEXIA treatment ,BIOPSY ,LEUCOCYTES ,SKELETAL muscle ,EXERCISE ,ANIMALS ,RATS ,EXPERIMENTAL design ,CELL culture ,ANIMAL experimentation ,RESEARCH methodology ,TUMORS ,CACHEXIA ,MUSCULAR atrophy ,BIOMARKERS ,DISEASE progression ,DISEASE complications - Abstract
Simple Summary: Currently, muscle testing remains underutilized in cancer cachexia research, since muscle biopsies are mainly performed at the end of experimental protocols. We aimed to introduce the concept of longitudinal muscle testing for cancer cachexia by initially demonstrating its feasibility and safety for the test subjects. Following that, we tested the hypothesis on a tumor-bearing rat model. Results were indicative of heterogeneity in cancer cachexia manifestation between different subjects and throughout the disease course. There is an abundance of researched pathways and mechanisms of cachexia, as well as interventions to target them. Thus, moving forward, developing biomarkers to suggest "what to target and when to do it" is essential. Sequential muscle biopsies can serve as a promising tool to guide personalized precision treatment for cancer cachexia, as well as to monitor the disease evolution and response to therapy. In the rapidly evolving landscape of cancer cachexia research, the development and refinement of diagnostic and predictive biomarkers constitute an ongoing challenge. This study aims to introduce longitudinal muscle biopsies as a potential framework for disease monitoring and treatment. The initial feasibility and safety assessment was performed for healthy mice and rats that received two consecutive muscle biopsies. The assessment was performed by utilizing three different tools. Subsequently, the protocol was also applied in leiomyosarcoma tumor-bearing rats. Longitudinal muscle biopsies proved to be a safe and feasible technique, especially in rat models. The application of this protocol to tumor-bearing rats further affirmed its tolerability and feasibility, while microscopic evaluation of the biopsies demonstrated varying levels of muscle atrophy with or without leukocyte infiltration. In this tumor model, sequential muscle biopsies confirmed the variability of the cancer cachexia evolution among subjects and at different time-points. Despite the abundance of promising cancer cachexia data during the past decade, the full potential of muscle biopsies is not being leveraged. Sequential muscle biopsies throughout the disease course represent a feasible and safe tool that can be utilized to guide precision treatment and monitor the response in cancer cachexia research. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
40. Onward Spread from Liver Metastases Is a Major Cause of Multi-Organ Metastasis in a Mouse Model of Metastatic Colon Cancer.
- Author
-
Wijler, Liza A., Viergever, Bastiaan J., Strating, Esther, van Schelven, Susanne J., Poghosyan, Susanna, Frenkel, Nicola C., te Rietmole, Hedy, Verheem, Andre, Raats, Danielle A. E., Borel Rinkes, Inne H. M., Hagendoorn, Jeroen, and Kranenburg, Onno
- Subjects
LIVER tumors ,BIOLOGICAL models ,MACROPHAGES ,RESEARCH funding ,DESCRIPTIVE statistics ,COLON tumors ,METASTASIS ,MICE ,LUMINESCENCE spectroscopy ,IMMUNOHISTOCHEMISTRY ,ANIMAL experimentation ,DATA analysis software ,DISEASE complications - Abstract
Simple Summary: The prognosis of patients with metastatic colon cancer remains very poor. A minority of patients, mostly those with liver-restricted disease, are eligible for intentionally curative surgery. In most cases, however, the presence of extra-hepatic metastases precludes curative treatment. The aim of this study was to gain insight into the processes governing multi-organ metastasis. We applied the microsurgical transplantation of mouse colon tumor organoids into the caecum or into the liver of syngeneic immunocompetent mice. Colon tumors growing in the liver seeded distant metastases to the lungs and to the peritoneal cavity more efficiently than those growing in the caecum. This was associated with the formation of hotspots of macrophage-surrounded vitronectin-positive blood vessels, specifically in liver tumors. Thus, 'onward spread' from liver metastases plays a major role in multi-organ metastasis, potentially through liver-specific vascular hotspots. The therapeutic targeting of these signals may help achieve the containment of the disease within the liver, thus preventing multi-organ metastasis. Colorectal cancer metastasizes predominantly to the liver but also to the lungs and the peritoneum. The presence of extra-hepatic metastases limits curative (surgical) treatment options and is associated with very poor survival. The mechanisms governing multi-organ metastasis formation are incompletely understood. Here, we tested the hypothesis that the site of tumor growth influences extra-hepatic metastasis formation. To this end, we implanted murine colon cancer organoids into the primary tumor site (i.e., the caecum) and into the primary metastasis site (i.e., the liver) in immunocompetent mice. The organoid-initiated liver tumors were significantly more efficient in seeding distant metastases compared to tumors of the same origin growing in the caecum (intra-hepatic: 51 vs. 40%, p = 0.001; peritoneal cavity: 51% vs. 33%, p = 0.001; lungs: 30% vs. 7%, p = 0.017). The enhanced metastatic capacity of the liver tumors was associated with the formation of 'hotspots' of vitronectin-positive blood vessels surrounded by macrophages. RNA sequencing analysis of clinical samples showed a high expression of vitronectin in liver metastases, along with signatures reflecting hypoxia, angiogenesis, coagulation, and macrophages. We conclude that 'onward spread' from liver metastases is facilitated by liver-specific microenvironmental signals that cause the formation of macrophage-associated vascular hotspots. The therapeutic targeting of these signals may help to contain the disease within the liver and prevent onward spread. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
41. Activation of Epstein–Barr Virus' Lytic Cycle in Nasopharyngeal Carcinoma Cells by NEO212, a Conjugate of Perillyl Alcohol and Temozolomide.
- Author
-
Hartman-Houstman, Hannah, Swenson, Steve, Minea, Radu O., Sinha, Uttam K., Chiang, Ming-Fu, Chen, Thomas C., and Schönthal, Axel H.
- Subjects
RISK assessment ,IN vitro studies ,RESEARCH funding ,EPSTEIN-Barr virus diseases ,TEMOZOLOMIDE ,BRAIN ,ENDOPLASMIC reticulum ,GANCICLOVIR ,IN vivo studies ,TUMOR markers ,DESCRIPTIVE statistics ,CELL lines ,METASTASIS ,MICE ,REACTIVE oxygen species ,ANIMAL experimentation ,NASOPHARYNX cancer ,ALCOHOLS (Chemical class) ,DISEASE risk factors ,DISEASE complications - Abstract
Simple Summary: Nasopharyngeal carcinoma (NPC) is a cancer that is frequently associated with Epstein–Barr virus (EBV) infection, and this virus is thought to play a role in NPC development and malignant growth. Within tumor cells, the virus generally remains in a latent stage that supports cell survival and stimulates tumor growth. However, it has been postulated that it might be beneficial as part of cancer therapy to coax the virus out of its latent stage and into a lytic mode, which would result in tumor cell killing. A few agents are known to accomplish this switch, but their clinical success so far has been mixed. We are developing a novel compound, NEO212, that has shown anticancer activity in several preclinical tumor models. Here, in this report, we demonstrate that it is able to coax EBV into its lytic cycle, resulting in NPC cell death. We propose that NEO212 should be investigated further for its potential clinical usefulness as a therapeutic agent for NPC. The Epstein–Barr virus (EBV) is accepted as a primary risk factor for certain nasopharyngeal carcinoma (NPC) subtypes, where the virus persists in a latent stage which is thought to contribute to tumorigenesis. Current treatments are sub-optimal, and recurrence occurs in many cases. An alternative therapeutic concept is aimed at triggering the lytic cycle of EBV selectively in tumor cells as a means to add clinical benefit. While compounds able to stimulate the lytic cascade have been identified, their clinical application so far has been limited. We are developing a novel anticancer molecule, NEO212, that was generated by covalent conjugation of the alkylating agent temozolomide (TMZ) to the naturally occurring monoterpene perillyl alcohol (POH). In the current study, we investigated its potential to trigger the lytic cycle of EBV in NPC cells in vitro and in vivo. We used the established C666.1 cell line and primary patient cells derived from the brain metastasis of a patient with NPC, both of which harbored latent EBV. Upon treatment with NEO212, there was an increase in EBV proteins Zta and Ea-D, key markers of the lytic cycle, along with increased levels of CCAAT/enhancer-binding protein homologous protein (CHOP), a marker of endoplasmic reticulum (ER) stress, followed by the activation of caspases. These effects could also be confirmed in tumor tissue from mice implanted with C666.1 cells. Towards a mechanistic understanding of these events, we used siRNA-mediated knockdown of CHOP and inclusion of anti-oxidant compounds. Both approaches blocked lytic cycle induction by NEO212. Therefore, we established a sequence of events, where NEO212 caused reactive oxygen species (ROS) production, which triggered ER stress and elevated the levels of CHOP, which was required to stimulate the lytic cascade of EBV. Inclusion of the antiviral agent ganciclovir synergistically enhanced the cytotoxic impact of NEO212, pointing to a potential combination treatment for EBV-positive cancers which should be explored further. Overall, our study establishes NEO212 as a novel agent able to stimulate EBV's lytic cycle in NPC tumors, with implications for other virus-associated cancers. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
42. Inhibition of TNBC Cell Growth by Paroxetine: Induction of Apoptosis and Blockage of Autophagy Flux.
- Author
-
Huang, Qianrui, Wu, Mengling, Pu, Yamin, Zhou, Junyou, Zhang, Yiqian, Li, Ru, Xia, Yong, Zhang, Yiwen, and Ma, Yimei
- Subjects
RNA analysis ,IN vitro studies ,FLOW cytometry ,BIOLOGICAL models ,AUTOPHAGY ,PROTEIN kinases ,BREAST tumors ,ANTINEOPLASTIC agents ,CHALONES ,APOPTOSIS ,IN vivo studies ,CELLULAR signal transduction ,CELL lines ,MICE ,DRUG repositioning ,ANIMAL experimentation ,WESTERN immunoblotting ,DOXORUBICIN ,PAROXETINE ,CELL survival ,SEQUENCE analysis ,PHARMACODYNAMICS - Abstract
Simple Summary: The objective of drug repurposing is to discover novel therapeutic uses for established pharmaceuticals. Paroxetine (PX), a commonly prescribed antidepressant, has demonstrated promising anticancer properties in experimental studies. Nevertheless, its precise role and mechanisms of action in triple-negative breast cancer (TNBC) remain incompletely elucidated. Our research findings propose that PX may impact TNBC by modulating critical molecular pathways, providing significant implications for enhancing chemosensitivity and identifying potential therapeutic synergies in clinical practice. These findings provide a basis for the creation of individualized treatment plans and the identification of the most effective therapeutic interventions to enhance treatment precision and effectiveness in patients with TNBC. The strategy of drug repurposing has gained traction in the field of cancer therapy as a means of discovering novel therapeutic uses for established pharmaceuticals. Paroxetine (PX), a selective serotonin reuptake inhibitor typically utilized in the treatment of depression, has demonstrated promise as an agent for combating cancer. Nevertheless, the specific functions and mechanisms by which PX operates in the context of triple-negative breast cancer (TNBC) remain ambiguous. This study aimed to examine the impact of PX on TNBC cells in vitro as both a standalone treatment and in conjunction with other pharmaceutical agents. Cell viability was measured using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, apoptosis was assessed through flow cytometry, and the effects on signaling pathways were analyzed using RNA sequencing and Western blot techniques. Furthermore, a subcutaneous tumor model was utilized to assess the in vivo efficacy of combination therapy on tumor growth. The results of our study suggest that PX may activate the Ca
2+ -dependent mitochondria-mediated intrinsic apoptosis pathway in TNBC by potentially influencing the PI3K/AKT/mTOR pathway as well as by inducing cytoprotective autophagy. Additionally, the combination of PX and chemotherapeutic agents demonstrated moderate inhibitory effects on 4T1 tumor growth in an in vivo model. These findings indicate that PX may exert its effects on TNBC through modulation of critical molecular pathways, offering important implications for improving chemosensitivity and identifying potential therapeutic combinations for clinical use. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
43. Identifying PLAUR as a Pivotal Gene of Tumor Microenvironment and Regulating Mesenchymal Phenotype of Glioblastoma.
- Author
-
Fu, Zaixiang, Chen, Zihang, Ye, Jingya, Ji, Jianxiong, Ni, Weifang, Lin, Weibo, Lin, Haopu, Lu, Liquan, Zhu, Ganggui, Xie, Qin, Yan, Feng, Chen, Gao, and Liu, Fuyi
- Subjects
BIOLOGICAL models ,KRUSKAL-Wallis Test ,ANIMAL experimentation ,ONE-way analysis of variance ,GLIOMAS ,MANN Whitney U Test ,GENES ,STEM cells ,DESCRIPTIVE statistics ,RESEARCH funding ,CELL lines ,DATA analysis software ,PHENOTYPES ,MICE - Abstract
Simple Summary: The mesenchymal (MES) phenotype of glioblastoma (GBM) confers resistance to various therapeutic strategies and results from both tumor-intrinsic genetic alterations and tumor-extrinsic microenvironmental factors. In this study, we sought to identify and validate the key genes that regulate the MES phenotype of glioma via both mechanisms. By integrating bulk tumor data (using the TCGA and CGGA databases) and single-cell sequencing data of GBM, we revealed that the plasminogen activator, urokinase receptor (PLAUR) is the hub gene of the protumor microenvironment, encompassing hypoxia and immunosuppression, and we elucidated its role in driving the MES transition through both tumor-intrinsic function and cell-to-cell interaction. Our findings indicate that PLAUR may be a potential target for GBM treatment, as our research elucidated its functions through a comprehensive study. The mesenchymal (MES) phenotype of glioblastoma (GBM) is the most aggressive and therapy-resistant subtype of GBM. The MES phenotype transition during tumor progression results from both tumor-intrinsic genetic alterations and tumor-extrinsic microenvironmental factors. In this study, we sought to identify genes that can modulate the MES phenotype via both mechanisms. By integrating weighted gene co-expression network analysis (WGCNA) and the differential expression analysis of hypoxia-immunosuppression-related genes, we identified the plasminogen activator, urokinase receptor (PLAUR) as the hub gene. Functional enrichment analysis and GSVA analysis demonstrated that PLAUR was associated with the MES phenotype of glioma and the hypoxia-immunosuppression-related microenvironmental components. Single-cell sequencing analysis revealed that PLAUR mediated the ligand–receptor interaction between tumor-associated macrophages (TAMs) and glioma cells. Functional experiments in vitro with cell lines or primary glioma cells and xenograft models using BALB/c nude mice confirmed the role of PLAUR in promoting the MES phenotype of GBM. Our findings indicate that PLAUR regulates both glioma cells and tumor cell-extrinsic factors that favor the MES phenotype and suggest that PLAUR might be a potential target for GBM therapy. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
44. RACK1 Promotes Meningioma Progression by Activation of NF-κB Pathway via Preventing CSNK2B from Ubiquitination Degradation.
- Author
-
Maalim, Ali Abdi, Wang, Zihan, Huang, Yimin, and Lei, Ting
- Subjects
BIOCHEMISTRY ,DISEASE progression ,STATISTICS ,IN vivo studies ,SEQUENCE analysis ,STAINS & staining (Microscopy) ,PHENOMENOLOGICAL biology ,ANIMAL experimentation ,COLONY-forming units assay ,WESTERN immunoblotting ,ONE-way analysis of variance ,CELL receptors ,NF-kappa B ,PRECIPITIN tests ,CELLULAR signal transduction ,CELL cycle ,MENINGIOMA ,TRANSFERASES ,MASS spectrometry ,GENE expression profiling ,CELL proliferation ,ENZYME-linked immunosorbent assay ,DESCRIPTIVE statistics ,RESEARCH funding ,CELL lines ,STATISTICAL correlation ,DATA analysis ,DATA analysis software ,MICE - Abstract
Simple Summary: Malignant meningiomas have high aggressiveness and recurrence rates and a poor prognosis, with no clear pharmacological treatment available. The study aims to investigate the progression mechanism of malignant meningiomas and the targets of intervention. RACK1 and CSNK2B have been shown to promote tumor progression but their roles in meningiomas are not clear. In this study, we will investigate the roles of RACK1-CSNK2B in meningiomas and search for targets to inhibit the progression of meningiomas. Higher-grade meningiomas (WHO grade II and III) are characterized by aggressive invasiveness and high postoperative recurrence rates. The prognosis remains inadequate even with adjuvant radiotherapy and currently there is no definitive pharmacological treatment strategy and target for malignant meningiomas. This study aims to unveil the mechanisms driving the malignant progression of meningiomas and to identify potential inhibitory targets, with significant clinical implications. Implementing techniques such as protein immunoprecipitation, mass spectrometry, RNA interference, and transcriptome sequencing, we investigated the malignancy mechanisms in meningioma cell lines IOMM-LEE and CH157-MN. Additionally, in vivo experiments were carried out on nude mice. We discovered a positive correlation between meningioma malignancy and the levels of the receptor for activated C kinase 1 (RACK1), which interacts with CSNK2B, the β subunit of casein kinase 2 (CK2), inhibiting its ubiquitination and subsequent degradation. This inhibition allows CK2 to activate the NF-κb pathway, which increases the transcription of CDK4 and cyclin D3, resulting in the transition of the cell cycle into the G2/M phase. The RACK1 inhibitor, harringtonolide (HA), significantly suppressed the malignant tendencies of meningioma cells. Our study suggests that RACK1 may play a role in the malignant progression of meningiomas, and therefore, targeting RACK1 could emerge as an effective strategy for reducing the malignancy of these tumors. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
45. ANLN Promotes the Proliferation and Migration of Gallbladder Cancer Cells via STRA6-Mediated Activation of PI3K/AKT Signaling.
- Author
-
Zhu, Xiang, Zhang, Yong, Bian, Rui, Zhu, Jiyue, Shi, Weibin, and Ye, Yuanyuan
- Subjects
REVERSE transcriptase polymerase chain reaction ,GALLBLADDER tumors ,IN vivo studies ,ONCOGENES ,ANIMAL experimentation ,WESTERN immunoblotting ,IMMUNOHISTOCHEMISTRY ,MICROFILAMENT proteins ,APOPTOSIS ,RNA ,CELL motility ,CELLULAR signal transduction ,CELL cycle ,BIOINFORMATICS ,CELL proliferation ,RESEARCH funding ,MEMBRANE proteins ,CELL lines - Abstract
Simple Summary: This study is the first to reveal the role and mechanism of ANLN (anillin, an actin-binding protein) in gallbladder cancer (GBC). Our research has led to the following conclusions: (1) ANLN promotes the proliferation and migration of gallbladder cancer cells. (2) Knockdown of ANLN promotes apoptosis and cell cycle arrest in GBC cells. (3) Knockdown of ANLN inhibits the Phosphatidylinositide 3-kinases (PI3K)/Serine/Threonine Kinase (AKT) signaling pathway, leading to the inhibition of GBC cell growth and migration. (4) ANLN activates the PI3K/AKT signaling pathway by regulating STRA6 in GBC cells. The ANLN gene encodes anillin, a protein that binds to actin. Recent research has identified ANLN's function in the initiation and advancement of different cancers. However, its impact on gallbladder cancer (GBC) remains unexplored. This study aimed to elucidate its possible molecular mechanisms in GBC. ANLN expression was assessed using quantitative real-time polymerase chain reaction (QRT-PCR), Western blotting (WB), and immunohistochemistry (IHC), revealing elevated levels in GBC tissues. ANLN knockdown resulted in the inhibition of cell proliferation and migration, leading to apoptosis and cell cycle arrest. Conversely, ANLN overexpression had the opposite effects on GBC cells. In vivo experiments confirmed that ANLN knockdown inhibited GBC cell growth. RNA-seq and bioinformatics analysis revealed ANLN's function in activating the PI3K/AKT signaling pathway. We further confirmed that ANLN could upregulate STRA6 expression, which activated PI3K/AKT signaling to enhance the growth and movement of GBC cells. These findings demonstrate ANLN's involvement in GBC initiation and progression, suggesting its potential as a novel target for GBC. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
46. Epigenetic Targeting to Overcome Radioresistance in Head and Neck Cancer.
- Author
-
Schniewind, Iñaki, Besso, Maria José, Klicker, Sebastian, Schwarz, Franziska Maria, Hadiwikarta, Wahyu Wijaya, Richter, Susan, Löck, Steffen, Linge, Annett, Krause, Mechthild, Dubrovska, Anna, Baumann, Michael, Kurth, Ina, and Peitzsch, Claudia
- Subjects
RNA analysis ,TRICARBOXYLIC acids ,FLOW cytometry ,BIOLOGICAL models ,SEQUENCE analysis ,XENOGRAFTS ,ANALYSIS of variance ,COLONY-forming units assay ,WESTERN immunoblotting ,CHEMILUMINESCENCE assay ,ANIMAL experimentation ,HEAD & neck cancer ,DNA methylation ,ALDEHYDE dehydrogenase ,T-test (Statistics) ,STEM cells ,MASS spectrometry ,CELL proliferation ,FLUORESCENT antibody technique ,GENE expression profiling ,DESCRIPTIVE statistics ,RESEARCH funding ,CELL lines ,DNA repair ,GENETIC techniques ,DATA analysis software ,TUMOR markers ,EPIGENOMICS ,SQUAMOUS cell carcinoma ,MICE - Abstract
Simple Summary: Cells with stem-like potential in head and neck squamous cell carcinoma (HNSCC) have been shown to exhibit features of intrinsic resistance to ionizing radiation. These cells, often referred to as cancer stem cells (CSCs), are characterized by an increased self-renewal and clonogenic potential, features associated with epithelial-to-mesenchymal transition (EMT), migratory behavior, and cellular plasticity. Cellular plasticity entails changes in phenotype and function in response to therapeutic pressures such as irradiation. Previously, we found that ALDH-positive CSCs are enriched in HNSCC cultures upon treatment with ionizing radiation. This process was characterized by mechanisms involving a selection of resistant populations and an induction of stem-like features. Cellular plasticity is regulated intracellularly through epigenetic mechanisms, including dynamic adaptations of DNA methylation and histone methylation, resulting in gene-specific alterations of expression levels. Therefore, we hypothesized that epigenetic targeting may prevent irradiation-induced cellular plasticity, rendering resistant HNSCC cells more sensitive. Employing a chemical library screen, we identified the histone demethylase inhibitor GSK-J1 to have a putative radiosensitizing effect that also reduces the stem-like potential of these cells. (1) Background: The sensitivity of head and neck squamous cell carcinoma (HNSCC) to ionizing radiation, among others, is determined by the number of cells with high clonogenic potential and stem-like features. These cellular characteristics are dynamically regulated in response to treatment and may lead to an enrichment of radioresistant cells with a cancer stem cell (CSC) phenotype. Epigenetic mechanisms, particularly DNA and histone methylation, are key regulators of gene-specific transcription and cellular plasticity. Therefore, we hypothesized that specific epigenetic targeting may prevent irradiation-induced plasticity and may sensitize HNSCC cells to radiotherapy. (2) Methods: We compared the DNA methylome and intracellular concentrations of tricarboxylic acid cycle metabolites in radioresistant FaDu and Cal33 cell lines with their parental controls, as well as aldehyde dehydrogenase (ALDH)-positive CSCs with negative controls. Moreover, we conducted a screen of a chemical library targeting enzymes involved in epigenetic regulation in combination with irradiation and analyzed the clonogenic potential, sphere formation, and DNA repair capacity to identify compounds with both radiosensitizing and CSC-targeting potential. (3) Results: We identified the histone demethylase inhibitor GSK-J1, which targets UTX (KDM6A) and JMJD3 (KDM6B), leading to increased H3K27 trimethylation, heterochromatin formation, and gene silencing. The clonogenic survival assay after siRNA-mediated knock-down of both genes radiosensitized Cal33 and SAS cell lines. Moreover, high KDM6A expression in tissue sections of patients with HNSCC was associated with improved locoregional control after primary (n = 137) and post-operative (n = 187) radio/chemotherapy. Conversely, high KDM6B expression was a prognostic factor for reduced overall survival. (4) Conclusions: Within this study, we investigated cellular and molecular mechanisms underlying irradiation-induced cellular plasticity, a key inducer of radioresistance, with a focus on epigenetic alterations. We identified UTX (KDM6A) as a putative prognostic and therapeutic target for HNSCC patients treated with radiotherapy. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
47. Hypoxia-Derived Exosomes Promote Lung Adenocarcinoma by Regulating HS3ST1-GPC4-Mediated Glycolysis.
- Author
-
Ji, Xianxiu, Zhu, Ren, Gao, Caixia, Xie, Huikang, Gong, Xiaomei, and Luo, Jie
- Subjects
RNA analysis ,ADENOCARCINOMA ,LUNG cancer ,REVERSE transcriptase polymerase chain reaction ,STATISTICS ,EXOSOMES ,XENOGRAFTS ,CELL culture ,STAINS & staining (Microscopy) ,CARCINOGENESIS ,ANIMAL experimentation ,COLONY-forming units assay ,WESTERN immunoblotting ,ONE-way analysis of variance ,PRECIPITIN tests ,APOPTOSIS ,METASTASIS ,CELL motility ,GLYCOSAMINOGLYCANS ,MEMBRANE glycoproteins ,CELL survival ,CELL cycle ,TRANSFERASES ,CELL proliferation ,DESCRIPTIVE statistics ,RESEARCH funding ,CELL lines ,TUMOR markers ,DATA analysis software ,DATA analysis ,HYPOXEMIA ,GLYCOLYSIS ,MICE - Abstract
Simple Summary: Lung cancer accounts for 11.4% of all cancer cases and 18.0% of all cancer deaths. Understanding the molecular mechanisms underlying lung adenocarcinoma (LUAD), one of the most common lung cancers, is imperative to enhance diagnostic accuracy, drug development, and prognostic evaluation. The present study explored the interaction between heparan sulfate (glucosamine) 3-O-sulfotransferase 1 (HS3ST1) and Glypican 4 (GPC4), as well as the influence of the hypoxia-derived exosomal lncRNA OIP5-AS1 on the glycolysis, proliferation, and metastasis ability in LUAD cell lines and their effects on the tumor growth in xenograft animal models. GPC4 promotes HS3ST1-mediated glycolysis, and the hypoxia-derived exosomal lncRNA OIP5-AS1 promotes glycolysis via the miR-200c-3p axis in LUAD cells. The hypoxia-derived exosomal lncRNA OIP5-AS1 enhances LUAD cell proliferation and metastasis in vitro and promotes LUAD tumor growth and metastasis via miR-200c-3p in vivo. These findings highlight that the hypoxia-derived exosomal lncRNA OIP5-AS1 may participate in LUAD progression. Objective: The diagnosis of lung adenocarcinoma (LUAD) is often delayed due to the typically asymptomatic nature of the early-stage disease, causing advanced-stage LUAD diagnosis in most patients. Hypoxia is widely recognized as a driving force in cancer progression. Exosomes originating from hypoxic tumor cells promote tumorigenesis by influencing glycolysis, migration, invasion, and immune infiltration. Given these insights, our study aimed to explore the role of hypoxia-derived exosomal long non-coding RNA (lncRNA) OIP5-AS1 in LUAD cell lines and mouse models. Materials and Methods: Exosomes were meticulously isolated and authenticated based on their morphology and biomarkers. The interaction between heparan sulfate (glucosamine) 3-O-sulfotransferase 1 (HS3ST1) and Glypican 4 (GPC4) was examined using immunoprecipitation. The influence of the hypoxia-derived exosomal lncRNA OIP5-AS1 on glycolysis was assessed in LUAD cell lines. The effect of the hypoxia-derived exosomal lncRNA OIP5-AS1 on cell proliferation and metastasis was evaluated using colony formation, cell viability, cell cycle, and apoptosis analyses. Its effects on tumor size were confirmed in xenograft animal models. Results: Our study revealed the mechanism of the hypoxia-derived exosomal lncRNA OIP5-AS1 in LUAD progression. We discovered that GPC4 promotes HS3ST1-mediated glycolysis and that the hypoxia-derived exosomal lncRNA OIP5-AS1 enhances glycolysis by regulating miR-200c-3p in LUAD cells. Notably, this lncRNA stimulates LUAD cell proliferation and metastasis and fosters LUAD tumor size via miR-200c-3p. Our findings underscore the potential role of the hypoxia-derived exosomal lncRNA OIP5-AS1 in LUAD progression. Conclusions: The hypoxia-derived exosomal lncRNA OIP5-AS1 promotes LUAD by regulating HS3ST1-GPC4-mediated glycolysis via miR-200c-3p. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
48. Examination of the Virulence of Actinobacillus pleuropneumoniae Serovar 16 in Pigs.
- Author
-
Tenk, Miklós, Tóth, Gergely, Márton, Zsuzsanna, Sárközi, Rita, Szórádi, Alejandra, Makrai, László, Pálmai, Nimród, Szalai, Tamás, Albert, Mihály, and Fodor, László
- Subjects
ACTINOBACILLUS pleuropneumoniae ,ACTINOBACILLUS ,ANIMAL herds ,SWINE ,SYMPTOMS ,ANIMAL experimentation ,PIGLETS - Abstract
Simple Summary: Actinobacillus (A.) pleuropneumoniae is a major agent of the porcine respiratory diseases complex (PRDC), together with several bacterium and virus species. A. pleuropneumoniae strains are diverse regarding virulence and antigen structure; they have different virulence variants, and the strains can be assigned into 19 serovars. Serovar 16 was described in Hungary for the first time, and it is relatively frequent in pig herds. The aim of the present work was examination of the virulence of the A. pleuropneumoniae A-85/14 strain, the type strain of serovar 16, by infecting piglets. The results prove that the A. pleuropneumoniae A-85/14 strain is a virulent one. Different virulence variants of A. pleuropneumoniae are involved in the etiology of porcine pleuropneumonia. The purpose of the present trial was examination of the virulence of the Actinobacillus pleuropneumoniae A-85/14 strain, the type strain of serovar 16, in an animal challenge experiment. Thirty 12-week-old piglets seronegative for A. pleuropneumoniae were allocated into three trial groups each of 10 animals, and they were infected intranasally with 10
6 , 107 , or 108 colony forming units (cfu) of the strain, respectively. Clinical signs were recorded twice a day, and the animals were euthanized 6 days after the infection. Typical clinical signs and postmortem lesions of porcine pleuropneumonia were seen in the animals of each trial group; however, they were generally mild, and no significant differences could be seen between the three groups. Even 106 colony forming units of A. pleuropneumoniae A-85/14 strain could induce clinical signs and lesions. Based on these results, the type strain of serovar 16 of A. pleuropneumoniae must be regarded as a typical pathogenic strain of the species. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
49. The Cellular and Organismal Effects of Nitroxides and Nitroxide-Containing Nanoparticles.
- Author
-
Sadowska-Bartosz, Izabela and Bartosz, Grzegorz
- Subjects
LONGEVITY ,ANIMAL experimentation ,ALKYL radicals ,IONIZING radiation ,NANOPARTICLES ,NITROXIDES - Abstract
Nitroxides are stable free radicals that have antioxidant properties. They react with many types of radicals, including alkyl and peroxyl radicals. They act as mimics of superoxide dismutase and stimulate the catalase activity of hemoproteins. In some situations, they may exhibit pro-oxidant activity, mainly due to the formation of oxoammonium cations as products of their oxidation. In this review, the cellular effects of nitroxides and their effects in animal experiments and clinical trials are discussed, including the beneficial effects in various pathological situations involving oxidative stress, protective effects against UV and ionizing radiation, and prolongation of the life span of cancer-prone mice. Nitroxides were used as active components of various types of nanoparticles. The application of these nanoparticles in cellular and animal experiments is also discussed. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
50. Macrophages Promote Subtype Conversion and Endocrine Resistance in Breast Cancer.
- Author
-
Zhang, Xiaoyan, Yang, Fengyu, Huang, Zhijian, Liu, Xiaojun, Xia, Gan, Huang, Jieye, Yang, Yang, Li, Junchen, Huang, Jin, Liu, Yuxin, Zhou, Ti, Qi, Weiwei, Gao, Guoquan, and Yang, Xia
- Subjects
ANIMAL experimentation ,MACROPHAGES ,LYMPH nodes ,METASTASIS ,CELL physiology ,BIOINFORMATICS ,GENOMICS ,RESEARCH funding ,TAMOXIFEN ,BREAST tumors ,MICE - Abstract
Simple Summary: Breast cancer is a highly heterogeneous disease on the molecular level. The molecular subtype will be altered during metastasis and affect patient outcomes and treatments. Breast cancer is prone to lymph node metastasis. Our study aimed to investigate whether subtype conversion occurs during lymph node metastasis and its underlying mechanism. We confirmed that hormone receptors were down-regulated, while HER2 was up-regulated during lymph node metastasis, and macrophages played a significant role in the process, probably via MNX1. Targeting macrophages or MNX1 may provide new avenues for endocrine therapy and targeted treatment of breast cancer patients with lymph node metastasis. Background: The progression of tumors from less aggressive subtypes to more aggressive states during metastasis poses challenges for treatment strategies. Previous studies have revealed the molecular subtype conversion between primary and metastatic tumors in breast cancer (BC). However, the subtype conversion during lymph node metastasis (LNM) and the underlying mechanism remains unclear. Methods: We compared clinical subtypes in paired primary tumors and positive lymph nodes (PLNs) in BC patients and further validated them in the mouse model. Bioinformatics analysis and macrophage-conditioned medium treatment were performed to investigate the role of macrophages in subtype conversion. Results: During LNM, hormone receptors (HRs) were down-regulated, while HER2 was up-regulated, leading to the transformation of luminal A tumors towards luminal B tumors and from luminal B subtype towards HER2-enriched (HER2-E) subtype. The mouse model demonstrated the elevated levels of HER2 in PLN while retaining luminal characteristics. Among the various cells in the tumor microenvironment (TME), macrophages were the most clinically relevant in terms of prognosis. The treatment of a macrophage-conditioned medium further confirmed the downregulation of HR expression and upregulation of HER2 expression, inducing tamoxifen resistance. Through bioinformatics analysis, MNX1 was identified as a potential transcription factor governing the expression of HR and HER2. Conclusion: Our study revealed the HER2-E subtype conversion during LNM in BC. Macrophages were the crucial cell type in TME, inducing the downregulation of HR and upregulation of HER2, probably via MNX1. Targeting macrophages or MNX1 may provide new avenues for endocrine therapy and targeted treatment of BC patients with LNM. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.