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Made available in DSpace on 2022-05-01T15:46:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) PIMREG expression strongly correlates with cellular proliferation in both malignant and normal cells. Throughout embryo development, PIMREG expression is prominent in the central nervous system. Recent studies have described elevated PIMREG expression in different types of tumors, which correlates with patient survival and tumor aggressiveness. Given the emerging significance of PIMREG in carcinogenesis and its putative role in the context of the nervous system, we investigated the expression and function of PIMREG in gliomas, the most common primary brain tumors. We performed an extensive analysis of PIMREG expression in tumors samples from glioma patients. We then assessed the effects of PIMREG silencing and overexpression on the sensitivity of glioblastoma cell lines treated with genotoxic agents commonly used for treating patients and assessed for treatment response, proliferation and migration. Our analysis shows that glioblastoma exhibits the highest levels of PIMREG expression among all cancers analyzed and that elevated PIMREG expression is a biomarker for glioma progression and patient outcome. Moreover, PIMREG is induced by genotoxic agents, and its silencing renders glioblastoma cells sensitive to temozolomide treatment and affects ATR- and ATM-dependent signaling. Our data demonstrate that PIMREG is involved in DNA damage response and temozolomide resistance of glioblastoma cells and further supports a role for PIMREG in tumorigenesis. Department of Cellular and Molecular Biology and Pathogenic Bioagents Ribeirão Preto Medical School University of São Paulo (FMRP-USP), São Paulo Department of Genetics Ribeirão Preto Medical School University of São Paulo (FMRP-USP), São Paulo School of Pharmaceutical Sciences São Paulo State University (UNESP), São Paulo School of Pharmaceutical Sciences São Paulo State University (UNESP), São Paulo FAPESP: 2019/26035-1

Made available in DSpace on 2020-12-12T01:45:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 This study aimed to identify and evaluate the cytotoxicity, genotoxicity, antigenotoxicity and chemoprevention assessment of flavonoids myricetin-3-O-(2″-O-galloyl)-α-rhamnopyranoside and myricetin-3-rhamnoside from Inga laurina leaves extract. The Quinone reductase induction as a biomarker for cancer chemoprevention was evaluated in murine hepatocellular carcinoma, the cytotoxicity was evaluated by sulforhonamide B assay and genotoxicity was evaluated by comet assay using HepG2 cell line. The results demonstrated that the flavonoids didn´t show cytotoxicity in HepG2 cells. In the chemoprevention evaluations were not able to promote the induction of Quinone Reductase and also no genotoxic effect was observed by evaluation of the comet assay in none of the concentrations tested. In the antigenotoxicity test, all compounds had a protective effect against damage induced by hydrogen peroxide and were repaired against damage. Although none of the flavonoids were capable of inducing the enzyme Quinone Reductase at the concentrations tested, the antigenotoxicity results showed a powerful chemoprotective action. School of Pharmaceutical Sciences Department of Clinical Analysis São Paulo State University UNESP Institute of Chemistry Nucleus Bioassays Biosynthesis & Ecophysiology of Natural Products São Paulo State University UNESP NIAC-Nucleus of Research Applied to Sciences-IFMA-Federal Institute of Education Science and Technology of Maranhão School of Pharmaceutical Sciences Department of Clinical Analysis São Paulo State University UNESP Institute of Chemistry Nucleus Bioassays Biosynthesis & Ecophysiology of Natural Products São Paulo State University UNESP

Made available in DSpace on 2021-06-25T11:18:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-05-25 Universidade Estadual Paulista Glioblastoma (GBM) is the most lethal and frequent type of brain tumor, leading patients to death in approximately 14 months after diagnosis. GBM treatment consists in surgical removal followed by radio and chemotherapy. However, tumors commonly relapse and the treatment promotes only a slight increase in patient survival. Thus, uncovering the cellular mechanisms involved in GBM resistance is of utmost interest, and the use of cell lines has been shown to be an extremely important tool. In this work, the exploration of RNAseq data from different GBM cell lines revealed different expression signatures, distinctly correlated with the behavior of GBM cell lines regarding proliferation indexes and radio-resistance. U87MG and U138MG cells, which presented expressively reduced proliferation and increased radio-resistance, showed a particular expression signature encompassing enrichment in many extracellular matrix (ECM) and receptor genes. Contrasting, U251MG and T98G cells, that presented higher proliferation and sensibility to radiation, exhibited distinct signatures revealing consistent enrichments for DNA repair processes and although several genes from the ECM-receptor pathway showed up-regulation, enrichments for this pathway were not detected. The ECM-receptor is a master regulatory pathway that is known to impact several cellular processes including: survival, proliferation, migration, invasion, and DNA damage signaling and repair, corroborating the associations we found. Furthermore, searches to The Cancer Genome Atlas (TCGA) repository revealed prognostic correlations with glioma patients for the majority of genes highlighted in the signatures and led to the identification of 31 ECM-receptor genes individually correlated with radiation responsiveness. Interestingly, we observed an association between the number of upregulated genes and survivability greater than 5 years after diagnosis, where almost all the patients that presented 21 or more upregulated genes were deceased before 5 years. Altogether our findings suggest the clinical relevance of ECM-receptor genes signature found here for radiotherapy decision and as biomarkers of glioma prognosis. School of Pharmaceutical Sciences São Paulo State University (UNESP) Center for Cell-Based Therapy (CTC) Regional Blood Center of Ribeirão Preto Department of Genetics Ribeirão Preto Medical School University of São Paulo (USP) Radiation Oncology Branch National Cancer Institute National Institutes of Health (NIH) School of Pharmaceutical Sciences São Paulo State University (UNESP)

Astrocytomas are the most common and aggressive type of primary brain tumors in adults. The World Health Organization (WHO) assorts them into grades, from I to IV, based on histopathological features that reflect their malignancy [1]. Alongside with tumor progression, comes an increased proliferation, genomic instability, infiltration in normal brain tissue and resistance to treatments. The high genomic instability forges tumor cells enhancing key proteins that avoid cells from collapsing and favor therapy resistance [2]. To explore genes and pathways associated with tumor progression phenotypes we analyzed gene expression in a panel of non-tumor and glioma cell lines, namely: ACBRI371, non-tumor human astrocytes; HDPC, fibroblasts derived from dental pulp; Res186, Res259, Res286 and UW467 that include grade I, II and III astrocytoma cell lines derived from pediatric tumors; and T98G, U343MG, U87MG, U138MG and U251MG, all derived from GBM (grade IV). We also profiled gene expression changes caused by exogenously induced replicative stress, performing RNA sequencing with camptothecin (CPT)-treated cells. Here we describe the RNA-sequencing data set acquired, including quality of reads and sequencing consistency, as well as the bioinformatics strategy used to analyze it. We also compared gene expression patterns and pathway enrichment between non-tumor versus lower-grade (LGG), non-tumor versus GBM, LGG versus GBM, and CPT-treated versus non-treated cells. In brief, a total of 6467 genes showed differential expression and 5 pathways were enriched in tumor progression, while 2279 genes and 7 pathways were altered under the replication stress condition. The raw data was deposited in the NCBI BioProject database under the accession number PRJNA631805. Our dataset is valuable for researchers interested in differential gene expression among different astrocytoma grades and in expression changes caused by replicative stress, facilitating studies that seek novel biomarkers of glioma progression and treatment resistance.

Made available in DSpace on 2020-12-10T19:49:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-02-12 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Procad Amazonia Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The objectives of this study were to extract and purify Bixin from the seeds of Bixa orellana and to evaluate its hypoglycemic activity in vivo, as well as, to conduct an in silico study of selectivity on peroxisome proliferator-activated receptors via molecular docking and molecular dynamics simulations. Oral administration of Bixin (10 mg/kg) significantly reduced their glucose level that was alloxan-induced diabetic rats. Bixin showed in silico selectivity on peroxisome proliferator-activated receptors (PPARs), particularly by the peroxisome proliferator-activated receptor gamma (PPAR gamma), which supports the hypoglycemic activity of Bixin. From the results obtained, it can be inferred that Bixin presents hypoglycemic characteristics, which was confirmed by the results obtained from the in vivo and in silico tests. Bixin may act by other pathways to control blood glucose and thus it is possible that it presents a different toxicity profile than troglitazone, rosiglitazone and pioglitazone. However, more studies on the activity and toxicity of Bixin are needed to evaluate for further clinical use. Univ Fed Amapa, Dept Biol Sci & Hlth, Lab Drugs Discovery, Macapa, Brazil Univ Sierra, Div Postgrade, Ixtlan De Juarez, Mexico Univ Fed Amapa, Dept Biol Sci & Hlth, Lab Modeling & Computat Chem, Macapa, Brazil Univ Fed Amapa, Res Grp Biocatalysis & Apllied Organ Synth, Macapa, Brazil Sao Paulo State Univ, Dept Nat Act Principles & Toxicol, Fac Pharmaceut Sci, Araraquara, SP, Brazil Univ Sao Paulo, Dept Cellular & Mol Biol, Ribeirao Preto Med Sch, Ribeirao Preto, Brazil Univ Nacl Autonoma Mexico, Fac Chem, Dept Pharm, Lab Pharmacol Nat Prod, Mexico City, DF, Mexico Univ Sao Paulo, Ribeirao Preto Med Sch, Bioinformat Grp, Dept Genet, Ribeirao Preto, Brazil Sao Paulo State Univ, Dept Nat Act Principles & Toxicol, Fac Pharmaceut Sci, Araraquara, SP, Brazil CAPES: PNPD/20130076-14001012005P1 Procad Amazonia: Auxpe - 1723/2018 CNPq: 407768/2013-0

The objectives of this study were to extract and purify Bixin from the seeds of

// Ozge Gursoy-Yuzugullu 1 , Chelsea Carman 1 , Rodolfo Bortolozo Serafim 1 , Marios Myronakis 1 , Valeria Valente 2 , Brendan D. Price 1 1 Department of Radiation Oncology, Dana-Farber Cancer Institute, Boston MA 02215, USA 2 Sao Paulo State University (UNESP), School of Pharmaceutical Sciences, Araraquara, Rodovia Araraquara-Jau, Campos Ville, SP, 14800-903, Brazil Correspondence to: Brendan D. Price, email: brendan_price@dfci.harvard.edu Keywords: DNA repair, radiosensitizer, glioma, histone methylation, G9a Received: January 04, 2017 Accepted: February 07, 2017 Published: February 20, 2017 ABSTRACT Radiation therapy is widely used to treat human malignancies, but many tumor types, including gliomas, exhibit significant radioresistance. Radiation therapy creates DNA double-strand breaks (DSBs), and DSB repair is linked to rapid changes in epigenetic modifications, including increased histone methylation. This increased histone methylation recruits DNA repair proteins which can then alter the local chromatin structure and promote repair. Consequently, combining inhibitors of specific histone methyltransferases with radiation therapy may increase tumor radiosensitivity, particularly in tumors with significant therapeutic resistance. Here, we demonstrate that inhibitors of the H4K20 methyltransferase SETD8 (UNC-0379) and the H3K9 methyltransferase G9a (BIX-01294) are effective radiosensitizers of human glioma cells. UNC-0379 blocked H4K20 methylation and reduced recruitment of the 53BP1 protein to DSBs, although this loss of 53BP1 caused only limited changes in radiosensitivity. In contrast, loss of H3K9 methylation through G9a inhibition with BIX-01294 increased radiosensitivity of a panel of glioma cells (SER2Gy range: 1.5 - 2.9). Further, loss of H3K9 methylation reduced DSB signaling dependent on H3K9, including reduced activation of the Tip60 acetyltransferase, loss of ATM signaling and reduced phosphorylation of the KAP-1 repressor. In addition, BIX-0194 inhibited DSB repair through both the homologous recombination and nonhomologous end-joining pathways. Inhibition of G9a and loss of H3K9 methylation is therefore an effective approach for increasing radiosensitivity of glioma cells. These results suggest that combining inhibitors of histone methyltransferases which are critical for DSB repair with radiation therapy may provide a new therapeutic route for sensitizing gliomas and other tumors to radiation therapy.

Made available in DSpace on 2020-12-12T02:36:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-05-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The Holliday Junction-Recognition Protein (HJURP) was reported as overexpressed in several cancers and also strongly correlated with poor prognosis of patients, especially in glioblastoma (GBM), the most common and deadly type of primary brain tumor. HJURP is responsible for loading the histone H3 variant—the Centromeric Protein A (CENP-A)—at the centromeres in a cell cycle-regulated manner, being required for proper chromosome segregation. Here we investigated HJURP association with survival and radioresistance of different GBM cell lines. HJURP knockdown compromised the clonogenic capacity and severely impaired survival of five distinct GBM cells, while nontumor astrocytes were not affected. U251MG cells showed a robust cell cycle arrest in G2/M phases followed by a drastic increment in cell death after HJURP silencing, while U138MG and U343MG cell lines presented augmented senescence with a comparable increase in cell death. Importantly, we verified that the impact on cell cycle dynamics and clonogenic survival were associated with loss CENP-A at the centromeres. Moreover, radiation resistance was also impacted by HJURP modulation in several GBM cell lines. U87MG, T98G, U138MG, and U343MG cells were all sensitized to ionizing radiation after HJURP reduction. These data reinforce the requirement of HJURP for proliferative capacity and radioresistance of tumor cells, underlining its potential as a promising therapeutic target for GBM. Department of Cellular and Molecular Biology Ribeirão Preto Medical School University of São Paulo (USP), Avenida Bandeirantes, 3900 São Paulo State University (UNESP) School of Pharmaceutical Sciences, Rodovia Araraquara - Jaú, Km 01 - s/n, Campos Ville Department of Genetics Ribeirão Preto Medical School University of São Paulo (USP), Avenida Bandeirantes, 3900 Center for Cell-Based Therapy-CEPID/FAPESP, Rua Tenente Catão Roxo, 2501 Center for Medical Genomics HCFMRP/USP and Center for Integrative System Biology - CISBi NAP/USP University of São Paulo Department of Radiation Oncology Dana-Farber Cancer Institute São Paulo State University (UNESP) School of Pharmaceutical Sciences, Rodovia Araraquara - Jaú, Km 01 - s/n, Campos Ville

Glioblastoma (GBM) is the most common and malignant type of primary brain tumor, showing rapid development and resistance to therapies. On average, patients survive 14.6 months after diagnosis and less than 5% survive five years or more. Several pieces of evidence have suggested that the DNA damage signaling and repair activities are directly correlated with GBM phenotype and exhibit opposite functions in cancer establishment and progression. The functions of these pathways appear to present a dual role in tumorigenesis and cancer progression. Activation and/or overexpression of ATRX, ATM and RAD51 genes were extensively characterized as barriers for GBM initiation, but paradoxically the exacerbated activity of these genes was further associated with cancer progression to more aggressive stages. Excessive amounts of other DNA repair proteins, namely HJURP, EXO1, NEIL3, BRCA2, and BRIP, have also been connected to proliferative competence, resistance and poor prognosis. This scenario suggests that these networks help tumor cells to manage replicative stress and treatment-induced damage, diminishing genome instability and conferring therapy resistance. Finally, in this review we address promising new drugs and therapeutic approaches with potential to improve patient survival. However, despite all technological advances, the prognosis is still dismal and further research is needed to dissect such complex mechanisms.

Made available in DSpace on 2019-10-06T16:23:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 This study aimed to evaluate the cytotoxicity and genotoxicity from Inga laurina leaves extracts and fractions and obtain their chemical profile. The chemical profile of the crude extract from I. laurina leaves and its fractions was investigated through 1 H NMR, RP-HPLC-PDA by co-injection with authentic standards and HPLC-MS. The quinone reductase induction as a biomarker for cancer chemoprevention was evaluated in murine hepatocellular carcinoma line, whereas the cytotoxicity was evaluated by sulforhodamine B assay (SRB) using HepG2 cell line and genotoxicity was evaluated by comet assay. The phytochemical analysis of the leaves crude extract and its fractions showed the presence of 2-hydroxyethyl-dodecanoate and the phenolic compounds: gallic acid, methyl gallate, p-coumaric acid, cinnamic acid, myricetin-3-O-(2″-O-galoyl)-α-rhamnopyranoside, proanthocyanidin A-2 and myricetrin. All the fractions tested were not considered cytotoxic against the selected human cancer cell lines, they did not cause genotoxic in some concentrations damage and induced the enzyme quinone reductase. School of Pharmaceutical Sciences Department of Clinical Analysis São Paulo State University UNESP Institute of Chemistry Nucleus Bioassays Biosynth & Ecophysiol Nat Prod São Paulo State University UNESP Department of Biotechnology Federal University of Amazonas University of Franca School of Pharmaceutical Sciences Department of Clinical Analysis São Paulo State University UNESP Institute of Chemistry Nucleus Bioassays Biosynth & Ecophysiol Nat Prod São Paulo State University UNESP

RMEL3 is a recently identified lncRNA associated with BRAFV600E mutation and melanoma cell survival. Here, we demonstrate strong and moderate RMEL3 upregulation in BRAF and NRAS mutant melanoma cells, respectively, compared to melanocytes. High expression is also more frequent in cutaneous than in acral/mucosal melanomas, and analysis of an ICGC melanoma dataset showed that mutations in RMEL3 locus are preponderantly C > T substitutions at dipyrimidine sites including CC > TT, typical of UV signature. RMEL3 mutation does not correlate with RMEL3 levels, but does with poor patient survival, in TCGA melanoma dataset. Accordingly, RMEL3 lncRNA levels were significantly reduced in BRAFV600E melanoma cells upon treatment with BRAF or MEK inhibitors, supporting the notion that BRAFMEK- ERK pathway plays a role to activate RMEL3 gene transcription. RMEL3 overexpression, in immortalized fibroblasts and melanoma cells, increased proliferation and survival under serum starvation, clonogenic ability, and xenografted melanoma tumor growth. Although future studies will be needed to elucidate the mechanistic activities of RMEL3, our data demonstrate that its overexpression bypasses the need of mitogen activation to sustain proliferation/survival of non-transformed cells and suggest an oncogenic role for RMEL3.

Multiple Sclerosis (MS) is an inflammatory neurodegenerative disease that affects approximately 2.5 million people globally. Even though the etiology of MS remains unknown, it is accepted that it involves a combination of genetic alterations and environmental factors. Here, after performing whole exome sequencing, we found a MS patient harboring a rare and homozygous single nucleotide variant (SNV; rs61745847) of the G-protein coupled receptor (GPCR) galanin-receptor 2 (GALR2) that alters an important amino acid in the TM6 molecular toggle switch region (W249L). Nuclear magnetic resonance imaging showed that the hypothalamus (an area rich in GALR2) of this patient exhibited an important volumetric reduction leading to an enlarged third ventricle. Ex vivo experiments with patient-derived blood cells (AKT phosphorylation), as well as studies in recombinant cell lines expressing the human GALR2 (calcium mobilization and NFAT mediated gene transcription), showed that galanin (GAL) was unable to stimulate cell signaling in cells expressing the variant GALR2 allele. Live cell confocal microscopy showed that the GALR2 mutant receptor was primarily localized to intracellular endosomes. We conclude that the W249L SNV is likely to abrogate GAL-mediated signaling through GALR2 due to the spontaneous internalization of this receptor in this patient. Although this homozygous SNV was rare in our MS cohort (1:262 cases), our findings raise the potential importance of impaired neuroregenerative pathways in the pathogenesis of MS, warrant future studies into the relevance of the GAL/GALR2 axis in MS and further suggest the activation of GALR2 as a potential therapeutic route for this disease.

CHD1 is a conserved chromatin remodeling enzyme required for development and linked to prostate cancer in adults, yet its role in human cells is poorly understood. Here, we show that targeted disruption of the CHD1 gene in human cells leads to a defect in early double-strand break (DSB) repair via homologous recombination (HR), resulting in hypersensitivity to ionizing radiation as well as PARP and PTEN inhibition. CHD1 knockout cells show reduced H2AX phosphorylation (γH2AX) and foci formation as well as impairments in CtIP recruitment to the damaged sites. Chromatin immunoprecipitation following a single DSB shows that the reduced levels of γH2AX accumulation at DSBs in CHD1-KO cells are due to both a global reduction in H2AX incorporation and poor retention of H2AX at the DSBs. We also identified a unique N-terminal region of CHD1 that inhibits the DNA binding, ATPase, and chromatin assembly and remodeling activities of CHD1. CHD1 lacking the N terminus was more active in rescuing the defects in γH2AX formation and CtIP recruitment in CHD1-KO cells than full-length CHD1, suggesting the N terminus is a negative regulator in cells. Our data point to a role for CHD1 in the DSB repair process and identify a novel regulatory region of the protein.

Made available in DSpace on 2018-12-11T17:31:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-02-10 Background: Photodynamic therapy (PDT) has proven to be a promising alternative to current cancer treatments, especially if combined with conventional approaches. The technique is based on the administration of a non-toxic photosensitizing agent to the patient with subsequent localized exposure to a light source of a specific wavelength, resulting in a cytotoxic response to oxidative damage. The present study intended to evaluate in vitro the type of induced death and the genotoxic and mutagenic effects of PDT alone and associated with cisplatin. Methods: We used the cell lines SiHa (ATCC® HTB35™), C-33 A (ATCC® HTB31™) and HaCaT cells, all available at Dr. Christiane Soares' Lab. Photosensitizers were Photogem (PGPDT) and methylene blue (MBPDT), alone or combined with cisplatin. Cell death was accessed through Hoechst and Propidium iodide staining and caspase-3 activity. Genotoxicity and mutagenicity were accessed via flow cytometry with anti-gama-H2AX and micronuclei assay, respectively. Data were analyzed by one-way ANOVA with Tukey's posthoc test. Results: Both MBPDT and PGPDT induced caspase-independent death, but MBPDT induced the morphology of typical necrosis, while PGPDT induced morphological alterations most similar to apoptosis. Cisplatin predominantly induced apoptosis, and the combined therapy induced variable rates of apoptosis- or necrosis-like phenotypes according to the cell line, but the percentage of dead cells was always higher than with monotherapies. MBPDT, either as monotherapy or in combination with cisplatin, was the unique therapy to induce significant damage to DNA (double strand breaks) in the three cell lines evaluated. However, there was no mutagenic potential observed for the damage induced by MBPDT, since the few cells that survived the treatment have lost their clonogenic capacity. Conclusions: Our results elicit the potential of combined therapy in diminishing the toxicity of antineoplastic drugs. Ultimately, photodynamic therapy mediated by either methylene blue or Photogem as monotherapy or in combination with cisplatin has low mutagenic potential, which supports its safe use in clinical practice for the treatment of cervical cancer. Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista (Unesp), Araraquara- Rod Araraquara-Jau km 01 s/n USP Univ de Sao Paulo Faculdade de Medicina de Ribeirao Preto, Avenida dos Bandeirantes 3900 Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista (Unesp), Araraquara- Rod Araraquara-Jau km 01 s/n

Made available in DSpace on 2022-04-29T08:45:04Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-04-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Astrocytomas are the most common primary brain tumors. They are very resistant to therapies and usually progress rapidly to high-grade lesions. Here, we investigated the potential role of DNA repair genes in astrocytoma progression and resistance. To this aim, we performed a polymerase chain reaction array-based analysis focused on DNA repair genes and searched for correlations between expression patters and survival prognoses. We found 19 genes significantly altered. Combining these genes in all possible arrangements, we found 421 expression signatures strongly associated with poor survival. Importantly, five genes (DDB2, EXO1, NEIL3, BRCA2, and BRIP1) were independently correlated with worse prognoses, revealing single-gene signatures. Moreover, silencing of EXO1, which is remarkably overexpressed, promoted faster restoration of double-strand breaks, while NEIL3 knockdown, also highly overexpressed, caused an increment in DNA damage and cell death after irradiation of glioblastoma cells. These results disclose the importance of DNA repair pathways for the maintenance of genomic stability of high-grade astrocytomas and suggest that EXO1 and NEIL3 overexpression confers more efficiency for double-strand break repair and resistance to reactive oxygen species, respectively. Thereby, we highlight these two genes as potentially related with tumor aggressiveness and promising candidates as novel therapeutic targets. Department of Clinical Analysis Faculty of Pharmaceutical Sciences of Araraquara University of São Paulo State Department of Cellular and Molecular Biology Ribeirão Preto Medical School University of São Paulo (USP) FAEPA Center for Medical Genomics (CMG) of the Clinical Hospital Ribeirão Preto Medical School University of São Paulo (USP) Department of Genetics Ribeirão Preto Medical School University of São Paulo (USP) Regional Blood Center of Ribeirão Preto and Center for Cell-Based Therapy—CEPID/FAPESP National Institute of Science and Technology in Stem cell and Cell Therapy Center for Integrative Systems Biology (CISBi) NAP/USP Department of Surgery and Anatomy Ribeirão Preto Medical School University of São Paulo (USP) Department of Pathology Ribeirão Preto Medical School University of São Paulo (USP) FAPESP: 2013/13465-1

The peroxisome proliferator-activated receptor gamma (PPARγ) is a nuclear receptor that acts as a transcription factor, regulating glucose, lipid and inflammation signaling and it is exploited in type 2 diabetes treatment. However, the selective activation of this PPAR subtype has been linked to important adverse effects which can be mitigated through concomitant activation of PPARα and PPARδ. In this study, we proposed new PPARγ agonists using PharmaGist Server for pharmacophore prediction, the molecular docking was performed by GOLD (genetic optimization for ligand docking) v2.2, AutoDock 4.2 and AutoDock Vina 1.1 and QikProp v4.0 and Derek for absorption, distribution, metabolism, excretion and toxicity (ADMET) assessment. One molecule showed high predicted affinity to PPARγ and favorable pharmacokinetic and toxicity properties. It was then evaluated against PPARα and PPARδ and showed greater affinity to these receptors than the controls. Therefore this molecule is a promising drug lead for the development of derivatives and for the treatment of metabolic syndrome with the benefits of a PPAR pan activation.

Made available in DSpace on 2015-03-18T15:56:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-11-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Studies of cell development in artificial microenvironments can contribute to understanding a series of physiological mechanisms that might be influenced by geometrical features of the microenvironment itself. In this work we applied two-photon polymerization to fabricate three-dimensional microenvironments composed of a matrix arrangement of microstructures (circular and square cross-sections pillars) with distinct spacing. Such microenvironments were used to evaluate the growth of Michigan Cancer Foundation-7 (MCF-7) cells that are commonly used as a model system to investigate fundamental aspects of the tumor biology. Our results reveal that the cell density decreases as the distance between structures in the environment is increased. Additionally, cell growth shows slightly better results for the microenvironments composed of circular cross-section structures. Univ Sao Paulo, Inst Fis Sao Carlos, BR-13560970 Sao Carlos, SP, Brazil Univ Estadual Paulista UNESP, Fac Ciencias Farmaceut, BR-14801960 Araraquara, SP, Brazil Univ Sao Paulo, Fac Med Ribeirao Preto, BR-14049900 Ribeirao Preto, SP, Brazil NAP USP, CISBi Ctr Integrat Syst Biol, Sao Paulo, Brazil Univ Estadual Paulista UNESP, Fac Ciencias Farmaceut, BR-14801960 Araraquara, SP, Brazil FAPESP: 11/12399-0 FAPESP: 12/21807-7

Made available in DSpace on 2014-12-03T13:11:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-04-25Bitstream added on 2014-12-03T13:22:30Z : No. of bitstreams: 1 WOS000318341400043.pdf: 5076716 bytes, checksum: 9511b533bf166bce399ed953ca17e01b (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundacao de Apoio ao Ensino, Pesquisa e Assistencia (FAEPA) do Hospital das Clinicas da Faculdade de Medicina de Ribeirao Preto Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Background: Diffuse astrocytomas are the most common type of primary brain cancer in adults. They present a wide variation in differentiation and aggressiveness, being classified into three grades: low-grade diffuse astrocytoma (grade II), anaplastic astrocytoma (grade III) and glioblastoma multiforme (grade IV), the most frequent and the major lethal type. Recent studies have highlighted the molecular heterogeneity of astrocytomas and demonstrated that large-scale analysis of gene expression could help in their classification and treatment. In this context, we previously demonstrated that HJURP, a novel protein involved in the repair of DNA double-strand breaks, is highly overexpressed in glioblastoma.Methodology/Principal Findings: Here we show that HJURP is remarkably overexpressed in a cohort composed of 40 patients with different grade astrocytomas. We also observed that tumors presenting the higher expression levels of HJURP are associated with poor survival prognosis, indicating HJURP overexpression as an independent prognostic factor of death risk for astrocytoma patients. More importantly, we found that HJURP knockdown strongly affects the maintenance of glioblastoma cells in a selective manner. Glioblastoma cells showed remarkable cell cycle arrest and premature senescence that culminated in elevated levels of cell death, differently from non-tumoral cells that were minimally affected.Conclusions: These data suggest that HJURP has an important role in the maintenance of extremely proliferative cells of high-grade gliomas and point to HJURP as a potential therapeutic target for the development of novel treatments for glioma patients. Univ Sao Paulo State UNESP, Fac Pharmaceut Sci Araraquara, Dept Clin Anal, Araraquara, Brazil NAP USP, Ctr Integrat Syst Biol CISBi, Ribeirao Preto, Brazil Barretos Canc Hosp, Mol Oncol Res Ctr, Barretos, Brazil Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Surg & Anat, Ribeirao Preto, Brazil Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Cellular & Mol Biol, Ribeirao Preto, Brazil Univ Sao Paulo, Fac Med, Dept Neurol, Ribeirao Preto, Brazil Univ Sao Paulo State UNESP, Fac Pharmaceut Sci Araraquara, Dept Clin Anal, Araraquara, Brazil FAPESP: 04/12133-6 FAPESP: 06/57602-9 FAPESP: 11/05674-4 CNPq: 485342/2006-5 CNPq: 154707/2006-6 CNPq: 19347/2011