17,070 results on '"Muscle proteins"'
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2. Information and control processes in living systems.
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Ramsey DM
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- Animals, California, Cell Differentiation, Central Nervous System physiology, Evoked Potentials, Group Processes, Models, Biological, Muscle Proteins, Nutritional Physiological Phenomena, Optic Nerve physiology, Vocalization, Animal, Biology, Cybernetics
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- 1968
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3. Effects of vigorous isometric muscle contraction on titin stiffness-related contractile properties in rat fast-twitch muscles
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Daiki Watanabe, Masanobu Wada, and Jiayu Shi
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Male ,Protein Kinase C-alpha ,Fast twitch muscle ,Physiology ,Muscle Proteins ,Isometric exercise ,Isometric Contraction ,Physiology (medical) ,medicine ,Animals ,Connectin ,Calcium Signaling ,Phosphorylation ,Rats, Wistar ,biology ,Muscle fatigue ,Calpain ,Chemistry ,Stiffness ,musculoskeletal system ,Electric Stimulation ,Isoenzymes ,Muscle Fatigue ,Muscle Fibers, Fast-Twitch ,Proteolysis ,biology.protein ,Biophysics ,Titin ,Isometric muscle contraction ,medicine.symptom - Abstract
This study was conducted to examine the effects of an acute bout of vigorous isometric contractions on titin stiffness-related contractile properties in rat fast-twitch skeletal muscles. Intact gastrocnemius muscles were electrically stimulated in situ until the force was reduced to ∼50% of the initial force. Immediately after cessation of the stimulation, the superficial regions of the muscles were dissected and subjected to biochemical and skinned fiber analyses. The stimulation resulted in a decrease in the titin-based passive force. The amounts of fragmented titin were unchanged by the stimulation. Protein kinase Cα-treatment increased the passive force in stimulated fibers to resting levels. The stimulation had no effect on the maximum Ca2+-activated force (max Ca2+ force) at a sarcomere length (SL) of 2.4 μm and decreased myofibrillar (my)-Ca2+ sensitivity at 2.6-μm SL. Stretching the SL to 3.0 μm led to the augmentation of the max Ca2+ force and my-Ca2+ sensitivity in both rested and stimulated fibers. For the max Ca2+ force, the extent of the increase was smaller in stimulated than in rested fibers, whereas for my-Ca2+ sensitivity, it was higher in stimulated than in rested fibers. These results suggest that vigorous isometric contractions decrease the titin-based passive force, possibly because of a reduction in phosphorylation by protein kinase Cα, and that the decreased titin stiffness may contribute, at least in part, to muscle fatigue. more...
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- 2021
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4. Novel role of prostate cancer risk variant rs7247241 on PPP1R14A isoform transition through allelic TF binding and CpG methylation
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Jong Y. Park, Yijun Tian, Alex Soupir, Qian Liu, Chiang Ching Huang, Lang Wu, and Liang Wang
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Male ,Bisulfite sequencing ,Muscle Proteins ,Single-nucleotide polymorphism ,Biology ,Polymorphism, Single Nucleotide ,Genetics ,Humans ,Protein Isoforms ,SNP ,Allele ,Molecular Biology ,Alleles ,Genetics (clinical) ,Intracellular Signaling Peptides and Proteins ,Prostatic Neoplasms ,RNA-Binding Proteins ,General Medicine ,Methylation ,DNA Methylation ,DNA-Binding Proteins ,CpG site ,CTCF ,DNA methylation ,Original Article ,CpG Islands ,Transcription Factors - Abstract
Although previous studies identified numerous single nucleotide polymorphisms (SNPs) and their target genes predisposed to prostate cancer (PrCa) risks, SNP-related splicing associations are rarely reported. In this study, we applied distance-based sQTL analysis (sQTLseekeR) using RNA-seq and SNP genotype data from benign prostate tissue (n = 467) and identified significant associations in 3344 SNP-transcript pairs (P ≤ 0.05) at PrCa risk loci. We characterized a common SNP (rs7247241) and its target gene (PPP1R14A) located in chr19q13, an sQTL with risk allele T associated with upregulation of long isoform (P = 9.99E−7). We confirmed the associations in both TCGA (P = 2.42E−24) and GTEX prostate cohorts (P = 9.08E−78). To functionally characterize this SNP, we performed chromatin immunoprecipitation qPCR and confirmed stronger CTCF and PLAGL2 binding in rs7247241 C than T allele. We found that CTCF binding enrichment was negatively associated with methylation level at the SNP site in human cell lines (r = −0.58). Bisulfite sequencing showed consistent association of rs7247241-T allele with nearby sequence CpG hypermethylation in prostate cell lines and tissues. Moreover, the methylation level at CpG sites nearest to the CTCF binding and first exon splice-in (ψ) of PPP1R14A was significantly associated with aggressive phenotype in the TCGA PrCa cohort. Meanwhile, the long isoform of the gene also promoted cell proliferation. Taken together, with the most updated gene annotations, we reported a set of sQTL associated with multiple traits related to human prostate diseases and revealed a unique role of PrCa risk SNP rs7247241 on PPP1R14A isoform transition. more...
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- 2021
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5. Intrinsic and acquired drug resistance to LSD1 inhibitors in small cell lung cancer occurs through a TEAD4‐driven transcriptional state
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Mark Ozeck, Timothy Nichols, Akshata Udyavar, Thomas A Paul, Tao Xie, Shikhar Sharma, Wen Yan, Jessica Frey, and Chi-Yeh Chung
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Cancer Research ,Lung Neoplasms ,animal structures ,Drug Resistance ,Muscle Proteins ,Drug resistance ,Neuroendocrine differentiation ,Genetics ,Humans ,TEAD4 ,Histone Demethylases ,biology ,TEA Domain Transcription Factors ,KDM1A ,General Medicine ,Small Cell Lung Carcinoma ,respiratory tract diseases ,DNA-Binding Proteins ,ASCL1 ,Oncology ,biology.protein ,Cancer research ,Molecular Medicine ,Demethylase ,Reprogramming ,Epigenetic therapy ,Transcription Factors - Abstract
Small cell lung cancer (SCLC) is a heterogeneous disease, consisting of intra-tumoral and inter-tumoral neuroendocrine (ASCL1 and/or NEUROD1), mesenchymal-like, and YAP-driven transcriptional states. Lysine-specific demethylase 1 (LSD1; also known as KDM1A) inhibitors have recently been progressed to clinical trials in SCLC based on a promising preclinical anti-tumor activity. A potential clinical limitation of LSD1 inhibitors is the heterogeneous drug responses that have been observed in SCLC cell lines and patient-derived models. Based on these observations, we studied molecular and transcriptional signatures that predict patient response to this class of drug. Employing SCLC patient-derived transcriptional signatures, we define that SCLC cell lines sensitive to LSD1 inhibitors are enriched in neuroendocrine transcriptional markers, whereas cell lines enriched in a mesenchymal-like transcriptional program demonstrate intrinsic resistance to LSD1 inhibitors. We have identified a reversible, adaptive resistance mechanism to LSD1 inhibitors through epigenetic reprogramming to a TEAD4-driven mesenchymal-like state. Our data suggest that only a segment of SCLC patients, with a defined neuroendocrine differentiation state, will likely benefit from LSD1 inhibitors. It provides novel evidence for the selection of a TEAD4-driven mesenchymal-like subpopulation resistant to LSD1 inhibitors in SCLC patients that may require effective drug combinations to sustain effective clinical responses. more...
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- 2021
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6. Estimating the age of Lucilia illustris during the intrapuparial period using two approaches: Morphological changes and differential gene expression.
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Wang, Yu, Gu, Zhi-ya, Xia, Shui-xiu, Wang, Jiang-feng, Zhang, Ying-na, and Tao, Lu-yang
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BLOWFLIES , *FORENSIC entomology , *INSECT morphology , *INSECT genetics , *GENE expression , *BIOLOGY , *FOOD habits , *FORENSIC sciences , *INSECTS , *HUMAN life cycle , *MUSCLE proteins , *POLYMERASE chain reaction , *POSTMORTEM changes , *PROTEINS - Abstract
Lucilia illustris (Meigen, 1826) (Diptera: Calliphoridae) is a cosmopolitan species of fly that has forensic and medical significance. However, there is no relevant study regarding the determination of the age of this species during the intrapuparial period. In this study, we investigated the changes in both morphology and differential gene expression during intrapuparial development, with an aim to estimate the age of L. illustris during the intrapuparial stage. The overall intrapuparial morphological changes of L. illustris were divided into 12 substages. Structures such as the compound eyes, mouthparts, antennae, thorax, legs, wings, and abdomen, each capable of indicating age during the intrapuparial stage, were observed in detail, and the developmental progression of each of these structures was divided into six to eight stages. We recorded the time range over which each substage or structure appeared. The differential expression of the three genes 15_2, actin, and tbp previously identified for predicting the timing of intrapuparial development was measured during L. illustris metamorphosis. The expression of these genes was quantified by real-time PCR, and the results revealed that these genes can be used to estimate the age of L. illustris during the intrapuparial period, as they exhibit regular changes and temperature dependence. This study provides an important basis for estimating the minimum postmortem interval (PMImin) in forensic entomology according to changes in intrapuparial development and differential gene expression. Furthermore, combination of the two approaches can generate a more precise PMImin than either approach alone. [ABSTRACT FROM AUTHOR] more...
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- 2018
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7. Single-Molecule Force Spectroscopy Studies of Missense Titin Mutations That Are Likely Causing Cardiomyopathy
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Jiahao Xia, Jiacheng Zuo, Hongbin Li, and Denghuang Zhan
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Mutation, Missense ,Cardiomyopathy ,Muscle Proteins ,Microscopy, Atomic Force ,medicine.disease_cause ,Electrochemistry ,medicine ,Humans ,Molecule ,Missense mutation ,Connectin ,General Materials Science ,Denaturation (biochemistry) ,Spectroscopy ,Mutation ,biology ,Chemistry ,Spectrum Analysis ,Force spectroscopy ,Surfaces and Interfaces ,Condensed Matter Physics ,medicine.disease ,Elasticity ,Folding (chemistry) ,biology.protein ,Biophysics ,Titin ,Cardiomyopathies - Abstract
The giant muscle protein titin plays important roles in heart function. Mutations in titin have emerged as a major cause of familial cardiomyopathy. Missense mutations have been identified in cardiomyopathy patients; however, it is challenging to distinguish disease-causing mutations from benign ones. Given the importance of titin mechanics in heart function, it is critically important to elucidate the mechano-phenotypes of cardiomyopathy-causing mutations found in the elastic I-band part of cardiac titin. Using single-molecule atomic force microscopy (AFM) and equilibrium chemical denaturation, we investigated the mechanical and thermodynamic effects of two missense mutations, R57C-I94 and S22P-I84, found in the elastic I-band part of cardiac titin that were predicted to be likely causing cardiomyopathy by bioinformatics analysis. Our AFM results showed that mutation R57C had a significant destabilization effect on the I94 module. R57C reduced the mechanical unfolding force of I94 by ∼30-40 pN, accelerated the unfolding kinetics, and decelerated the folding. These effects collectively increased the unfolding propensity of I94, likely resulting in altered titin elasticity. In comparison, S22P led to only modest destabilization of I84, with a decrease in unfolding force by ∼10 pN. It is unlikely that such a modest destabilization would lead to a change in titin elasticity. These results will serve as the first step toward elucidating mechano-phenotypes of cardiomyopathy-causing mutations in the elastic I-band. more...
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- 2021
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8. Calpain-2 specifically cleaves Junctophilin-2 at the same site as Calpain-1 but with less efficacy
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E. Dale Abel, Qian Shi, Biyi Chen, Jinxi Wang, Tianqing Peng, Long-Sheng Song, Grace Ciampa, Dong Zheng, and Duane D. Hall
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Male ,Threonine ,Gene isoform ,Arginine ,Proteolysis ,Protein subunit ,Glycine ,Muscle Proteins ,Calpain-1 ,Transfection ,Cleavage (embryo) ,Biochemistry ,Mice ,medicine ,Animals ,Humans ,Myocytes, Cardiac ,Molecular Biology ,Research Articles ,Heart Failure ,medicine.diagnostic_test ,biology ,Calpain ,Chemistry ,Membrane Proteins ,Cell Biology ,Junctophilin-2 ,Deletion Mutagenesis ,Cell biology ,Disease Models, Animal ,HEK293 Cells ,Cardiovascular System & Vascular Biology ,Calpain-2 ,Mutagenesis, Site-Directed ,biology.protein ,Signal Transduction - Abstract
Calpain proteolysis contributes to the pathogenesis of heart failure but the calpain isoforms responsible and their substrate specificities have not been rigorously defined. One substrate, Junctophilin-2 (JP2), is essential for maintaining junctional cardiac dyads and excitation-contraction coupling. We previously demonstrated that mouse JP2 is cleaved by calpain-1 (CAPN1) between Arginine 565 (R565) and Threonine 566 (T566). Recently, calpain-2 (CAPN2) was reported to cleave JP2 at a novel site between Glycine 482 (G482) and Threonine 483 (T483). We aimed to directly compare the contributions of each calpain isoform, their Ca2+ sensitivity, and their cleavage site selection for JP2. We find CAPN1, CAPN2 and their requisite CAPNS1 regulatory subunit are induced by pressure overload stress that is concurrent with JP2 cleavage. Using in vitro calpain cleavage assays, we demonstrate that CAPN1 and CAPN2 cleave JP2 into similar 75 kD N-terminal (JP2NT) and 25 kD C-terminal fragments (JP2CT) with CAPNS1 co-expression enhancing proteolysis. Deletion mutagenesis shows both CAPN1 and CAPN2 require R565/T566 but not G482/T483. When heterologously expressed, the JP2CT peptide corresponding to R565/T566 cleavage approximates the 25 kD species found during cardiac stress while the C-terminal peptide from potential cleavage at G482/T483 produces a 35 kD product. Similar results were obtained for human JP2. Finally, we show that CAPN1 has higher Ca2+ sensitivity and cleavage efficacy than CAPN2 on JP2 and other cardiac substrates including cTnT, cTnI and β2-spectrin. We conclude that CAPN2 cleaves JP2 at the same functionally conserved R565/T566 site as CAPN1 but with less efficacy and suggest heart failure may be targeted through specific inhibition of CAPN1. more...
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- 2021
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9. Transgelin-2 interacts with CD44 to regulate Notch1 signaling pathway and participates in colorectal cancer proliferation and migration
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Ming Huang, Yunfeng Li, Yi Gao, Lijuan Zhang, Tao Shen, Xuan Zhang, Rong Ding, Yueyi Yao, Jiayi Li, Tao Wu, Shuzhen Kong, and Guoyu Li
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Physiology ,Colorectal cancer ,Transgelin ,Muscle Proteins ,Biochemistry ,Cell Movement ,Cell Line, Tumor ,Overall survival ,medicine ,Humans ,Notch1 signaling ,Receptor, Notch1 ,neoplasms ,Cell Proliferation ,Gene knockdown ,biology ,Microfilament Proteins ,CD44 ,General Medicine ,medicine.disease ,digestive system diseases ,Hyaluronan Receptors ,Molecular mechanism ,biology.protein ,Cancer research ,Signal transduction ,Colorectal Neoplasms ,Signal Transduction - Abstract
The abnormal expression of transgelin-2 (TAGLN2) is related to tumor occurrence and progression. However, the underlying molecular mechanism of TAGLN2 in human colorectal cancer (CRC) is still poorly understood. Compared with adjacent tissues, TAGLN2 is overexpressed in CRC tissues. Its expression level is negatively correlated with the overall survival rate of patients with CRC. In addition, knockdown of TAGLN2 inhibited the proliferation and invasion of CRC cells. We also showed that TAGLN2 could interact with CD44 to regulate the Notch-1 signaling pathway. Our findings indicate there is increased TAGLN2 expression in CRC and that it may serve as a promising potential therapeutic target for CRC. more...
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- 2021
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10. MiR-29ab1 Cluster Resists Muscle Atrophy Through Inhibiting MuRF1
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Guangbin Zhou, Mengxu Ge, Bing Zhang, Qingyong Meng, Miaomiao Lan, Yang Su, Yuying Zhang, Lijie Gu, Chang Liu, Yingying Yu, Lei Li, Tongtong Wang, Kuo Zhang, and Liu Chuncheng
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Ubiquitin-Protein Ligases ,Muscle Proteins ,Biology ,Protein degradation ,Muscle Development ,Myoblasts ,Tripartite Motif Proteins ,Mice ,Atrophy ,Downregulation and upregulation ,Genetics ,medicine ,Animals ,Humans ,Muscle, Skeletal ,Molecular Biology ,Mice, Knockout ,Denervation ,Myogenesis ,Skeletal muscle ,Cell Biology ,General Medicine ,medicine.disease ,Muscle atrophy ,Cell biology ,MicroRNAs ,Muscular Atrophy ,HEK293 Cells ,medicine.anatomical_structure ,Knockout mouse ,medicine.symptom - Abstract
Skeletal muscle has great plasticity. An increase in protein degradation can cause muscle atrophy. Atrogin-1 and muscle ring finger-1 (MuRF1) are dramatically upregulated in various muscle atrophy. Inhibition of Atrogin-1 and MuRF1 protects against muscle atrophy. MiR-29 plays an important regulatory role in skeletal muscle development. However, the function of miR-29 in skeletal muscle protein metabolism is not clear. To investigate the function of miR-29, we generated miR-29 knockout mice and the miR-29ab1 cluster overexpression mice. The disruption of miR-29 led to severe atrophy of skeletal muscle during puberty, and the muscle-specific overexpression of the miR-29ab1 cluster protected against denervation-induced and fasting-induced muscle atrophy. Furthermore, the overexpression of miR-29a, b mimics in myotubes resisted the muscle atrophy. MuRF1 was the direct target gene of miR-29a, b. These results demonstrate that miR-29ab1 cluster plays a critical role in the maintenance of skeletal muscle. MiR-29ab1 cluster is the excellent inhibitor of MuRF1, ultimately indicating that miR-29ab1 cluster is good therapeutic molecule candidate for adulthood. more...
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- 2021
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11. Reducing body myopathy – A new pathogenic FHL1 variant and literature review
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Isabella Araújo Mota, Pedro Nogueira Fontana, Alzira Alves de Siqueira Carvalho, and Carolina da Cunha Correia
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Adult ,Male ,0301 basic medicine ,Pediatrics ,medicine.medical_specialty ,Adolescent ,Biopsy ,Mutation, Missense ,Muscle Proteins ,Reducing body myopathy ,Young Adult ,03 medical and health sciences ,0302 clinical medicine ,Muscular Diseases ,medicine ,Humans ,Genetic Predisposition to Disease ,Genetic Testing ,Child ,Muscle, Skeletal ,Genetics (clinical) ,Muscle biopsy ,medicine.diagnostic_test ,biology ,business.industry ,Intracellular Signaling Peptides and Proteins ,Genetic Diseases, X-Linked ,Progressive muscle weakness ,LIM Domain Proteins ,Middle Aged ,FHL1 ,Pedigree ,Phenotype ,030104 developmental biology ,Neurology ,Severe phenotype ,Child, Preschool ,Mutation ,Pediatrics, Perinatology and Child Health ,biology.protein ,Female ,Creatine kinase ,Neurology (clinical) ,business ,030217 neurology & neurosurgery ,Progressive muscular weakness ,Rare disease - Abstract
Reducing body myopathy (RBM) is a rare disease marked by progressive muscle weakness caused by a mutation in FHL1 gene. We describe a new pathogenic variant and contrasted it with 44 other cases identified in the literature. A male child presented at age 3 suffering frequent falls and progressive muscular weakness. At age 8, he was wheelchair-bound and required ventilatory support. His mother and sister died due to the same problem. Creatine kinase was 428 IU/L (190). Muscle biopsy showed typical reducing bodies, and genetic analysis identified a novel pathogenic hemizygous variant, c.370_375del. We identified 44 previous reported cases separated in two groups: 28 cases with mean age onset 7.6 ± 5 years and 16 with 26.7 ± 4.2 years. The time for the diagnosis was shorter to younger group. The initial symptoms, rigid spine, contractures, scoliosis and axial and neck weaknesses, dysphagia, cardiac involvement, were predominant in younger group. The variant c.369C G predominated in younger group and c.448T C in older one. Pathogenic variants positions seemed related to severe phenotype. Most wheelchair patients belonged to younger group. The data from this compilation and our case provided a general characterization spectrum and prognosis between two groups of age onset with RBM. more...
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- 2021
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12. TAGLN mediated stiffness-regulated ovarian cancer progression via RhoA/ROCK pathway
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Xiao Wei, Chaoyang Sun, Jingjing Ma, Dongchen Zhou, Hua Lou, Sen Xu, Qinglei Gao, Teng Ji, Xiaoting Li, Zongyuan Yang, Yijuan Jia, Xin Yang, Quanfu Huang, Zhongzhen Guo, Huayi Li, and Yunchong Meng
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Cancer Research ,RHOA ,Fluorescent Antibody Technique ,Gene Expression ,Muscle Proteins ,Mice ,Tumor Microenvironment ,Medicine ,Neoplasm Metastasis ,RC254-282 ,Ovarian Neoplasms ,rho-Associated Kinases ,medicine.diagnostic_test ,biology ,Progression ,Chemistry ,Microfilament Proteins ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Prognosis ,Extracellular Matrix ,src-Family Kinases ,Oncology ,Disease Progression ,Immunohistochemistry ,Female ,medicine.symptom ,Proto-oncogene tyrosine-protein kinase Src ,Signal Transduction ,TAGLN ,Immunofluorescence ,Models, Biological ,Tissue mechanics ,Western blot ,In vivo ,Ovarian cancer ,Cell Line, Tumor ,Animals ,Humans ,business.industry ,Research ,Gene Expression Profiling ,medicine.disease ,Desmoplasia ,Disease Models, Animal ,Cancer research ,biology.protein ,RhoA/ROCK ,business ,rhoA GTP-Binding Protein - Abstract
Background: Ovarian cancer (OC) progression is an unmet medical challenge. Since omental metastases were palpated harder than their primary counterparts during cytoreductive surgery of patients with epithelial ovarian cancer, we were inspired to investigate OC progression from the perspective of biomechanics. Methods: Atomic Force Microscope (AFM) was used to measure the young’s moduli of tissues. The collagen-coated polyacrylamide hydrogel (PA gel) system was prepared to mimic the soft and stiff substrates in vitro. The effect of TAGLN were evaluated both in vitro and in vivo using transwell, immunofluorescence, western blot analysis and immunohistochemistry. Findings: We quantitatively confirmed that omental metastases were stiffer and more abundant in desmoplasia compared with paired primary tumors, and further demonstrated that matrix stiffness could notably regulate OC progression. Remarkably, TAGLN, encoding an actin cross-linking/gelling protein, was identified as a potent mechanosensitive gene that could form a regulation loop with Src activation reacting to environmental stiffness, thus mediating stiffness-regulated OC progression through regulating RhoA/ROCK pathway. Interpretation: These data demonstrate that targeting extra-cellular matrix stiffness could probably hamper OC progression, and of note, targeting TAGLN might provide promising clinical therapeutic value for OC therapy. Funding: This work was supported by the “973” Program of China (No. 2015CB553903 to Ding Ma and Junbo Hu), Technical Innovation Special Project of Hubei Province (2018ACA138), the Fundamental Research Funds for the Central Universities (2019kfyXMBZ024), National Science and Technology Major Sub- Project (2018ZX10301402-002), and the National Science Foundation of China (81902661, 81772787, 82072889). Declaration of Interest: None to declare. Ethical Approval: Human ovarian cancer tissues were all obtained from patients undergoing surgery at the Department of Obstetrics and Gynecology, Tongji Hospital, Huazhong University of Science and Technology after obtaining written informed consent of the patients and the authorization of the Ethics Committee of Tongji Hospital (TJ-iRB20181103). Fresh tissues were prepared for the Atomic Force Microscope (AFM) measurement, frozen or formalin fixed. more...
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- 2021
13. Potential DPP IV Inhibitory Peptides from Dry-Cured Pork Loins after Hydrolysis: An In Vitro and In Silico Study
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Joanna Stadnik and Paulina Kęska
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Microbiology (medical) ,QH301-705.5 ,In silico ,Muscle Proteins ,Chemical Fractionation ,dry-cured meat ,Microbiology ,Structure-Activity Relationship ,Hydrolysis ,Pepsin ,Protein Interaction Mapping ,spectrometric analysis ,Amino Acid Sequence ,Protein Interaction Maps ,Biology (General) ,Molecular Biology ,chemistry.chemical_classification ,Myomesin ,Dipeptidyl-Peptidase IV Inhibitors ,biology ,Chemistry ,Spectrum Analysis ,General Medicine ,Metabolism ,In vitro ,Enzyme ,DPP-IV ,Biochemistry ,Toxicity ,Pork Meat ,biology.protein ,peptides ,Protein Binding - Abstract
Peptidyl peptidase IV (DPP-IV) is a pharmacotherapeutic target in type 2 diabetes, and inhibitors of this enzyme are an important class of drugs for the treatment of type 2 diabetes. In the present study, peptides (<, 7 kDa) isolated from dry-cured pork loins after pepsin and pancreatin hydrolysis were identified by mass spectrometry and tested as potential inhibitors of DPP-IV by the in silico method. Two peptides, namely WTIAVPGPPHS from myomesin (water-soluble fraction, A = 0.9091) and FKRPPL from troponin (salt-soluble fraction, A = 0.8333), were selected as the most promising inhibitors of DPP-IV. Both peptides were subjected to ADMET analysis. Fragments of these peptides showed promising drug-likeness properties as well as favorable absorption, distribution, metabolism, excretion, and toxicity functions, suggesting that they are novel leads in the development of DPP-IV inhibitors from food. more...
- Published
- 2021
14. Four-and-a-half LIM domain protein 2 (FHL2) deficiency protects mice from diet-induced obesity and high FHL2 expression marks human obesity
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Patrick C.N. Rensen, Carlie J.M. de Vries, Jayron J. Habibe, Sander Kooijman, Hilde Herrema, Mariska Vos, Maria P. Clemente-Olivo, Roelof Ottenhoff, Max Nieuwdorp, Noam Zelcer, Aldo Jongejan, Etto C. Eringa, Daniël H. van Raalte, Medical Biochemistry, ACS - Atherosclerosis & ischemic syndromes, ACS - Diabetes & metabolism, Amsterdam Gastroenterology Endocrinology Metabolism, Epidemiology and Data Science, APH - Methodology, Experimental Vascular Medicine, Vascular Medicine, APH - Personalized Medicine, ACS - Heart failure & arrhythmias, VU University medical center, Internal medicine, AGEM - Endocrinology, metabolism and nutrition, and Physiology more...
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Male ,0301 basic medicine ,Endocrinology, Diabetes and Metabolism ,Glucose uptake ,Muscle Proteins ,White adipose tissue ,Weight Gain ,Mice ,0302 clinical medicine ,Endocrinology ,Brown adipose tissue ,Mice, Knockout ,Adipogenesis ,White adipose tissue (WAT) ,Middle Aged ,medicine.anatomical_structure ,Female ,Signal transduction ,medicine.symptom ,Adult ,Browning of WAT ,medicine.medical_specialty ,Adipose Tissue, White ,LIM-Homeodomain Proteins ,030209 endocrinology & metabolism ,Biology ,Diet, High-Fat ,03 medical and health sciences ,Internal medicine ,medicine ,Animals ,Humans ,Genetic Predisposition to Disease ,Obesity ,Aged ,Wild type ,Lipid uptake ,Energy metabolism ,medicine.disease ,Diet-induced obese mice ,FHL2 ,Mice, Inbred C57BL ,030104 developmental biology ,Four-and-a-half LIM domain protein 2 (FHL2) ,Energy expenditure ,Weight gain ,Biomarkers ,Transcription Factors - Abstract
Objective: Four-and-a-Half-LIM-domain-protein 2 (FHL2) modulates multiple signal transduction pathways but has not been implicated in obesity or energy metabolism. In humans, methylation and expression of the FHL2 gene increases with age, and high FHL2 expression is associated with increased body weight in humans and mice. This led us to hypothesize that FHL2 is a determinant of diet-induced obesity. Methods: FHL2-deficient (FHL2 & minus;/& minus;) and wild type male mice were fed a high-fat diet. Metabolic phenotyping of these mice, as well as transcriptional analysis of key metabolic tissues was performed. Correlation of the expression of FHL2 and relevant genes was assessed in datasets from white adipose tissue of individuals with and without obesity. Results: FHL2 Deficiency protects mice from high-fat diet-induced weight gain, whereas glucose handling is normal. We observed enhanced energy expenditure, which may be explained by a combination of changes in multiple tissues; mild activation of brown adipose tissue with increased fatty acid uptake, increased cardiac glucose uptake and browning of white adipose tissue. Corroborating our findings in mice, expression of FHL2 in human white adipose tissue positively correlates with obesity and negatively with expression of browning-associated genes. Conclusion: Our results position FHL2 as a novel regulator of obesity and energy expenditure in mice and human. Given that FHL2 expression increases during aging, we now show that low FHL2 expression associates with a healthy metabolic state. (c) 2021 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY license (http:// creativecommons.org/licenses/by/4.0/). more...
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- 2021
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15. Recent advances in measuring and understanding the regulation of exercise-mediated protein degradation in skeletal muscle
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Yu-Chiang Lai, Yusuke Nishimura, Lars Holm, and Ibrahim Musa
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0301 basic medicine ,Proteasome Endopeptidase Complex ,Physiology ,Muscle Proteins ,Protein degradation ,Muscle mass ,03 medical and health sciences ,0302 clinical medicine ,Ubiquitin ,medicine ,Animals ,Humans ,Exercise physiology ,Muscle, Skeletal ,Exercise ,biology ,Chemistry ,Protein turnover ,Skeletal muscle ,Cell Biology ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Protein Biosynthesis ,biology.protein ,Phosphorylation ,Protein quality ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
Skeletal muscle protein turnover plays a crucial role in controlling muscle mass and protein quality control, including sarcomeric (structural and contractile) proteins. Protein turnover is a dynamic and continual process of protein synthesis and degradation. The ubiquitin proteasome system (UPS) is a key degradative system for protein degradation and protein quality control in skeletal muscle. UPS-mediated protein quality control is known to be impaired in aging and diseases. Exercise is a well-recognized, nonpharmacological approach to promote muscle protein turnover rates. Over the past decades, we have acquired substantial knowledge of molecular mechanisms of muscle protein synthesis after exercise. However, there have been considerable gaps in the mechanisms of how muscle protein degradation is regulated at the molecular level. The main challenge to understand muscle protein degradation is due in part to the lack of solid stable isotope tracer methodology to measure muscle protein degradation rate. Understanding the mechanisms of UPS with the concomitant measurement of protein degradation rate in skeletal muscle will help identify novel therapeutic strategies to ameliorate impaired protein turnover and protein quality control in aging and diseases. Thus, the goal of this present review was to highlight how recent advances in the field may help improve our understanding of exercise-mediated protein degradation. We discuss 1) the emerging roles of protein phosphorylation and ubiquitylation modifications in regulating proteasome-mediated protein degradation after exercise and 2) methodological advances to measure in vivo myofibrillar protein degradation rate using stable isotope tracer methods. more...
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- 2021
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16. Identification of New Transcription Factors that Can Promote Pluripotent Reprogramming
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Ping Zhu, Dongwei Li, Ping Huang, Yu Liu, Duanqing Pei, Ran Zhang, Jieying Zhu, and Guihuan Liu
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Induced Pluripotent Stem Cells ,Muscle Proteins ,Myocardial differentiation ,Biology ,Article ,SOX2 ,Humans ,TEAD4 ,TEAD2 ,Induced pluripotent stem cell ,Transcription factor ,Embryonic Stem Cells ,TEA Domain Transcription Factors ,Reprogramming ,Cell Differentiation ,Cellular Reprogramming ,Embryonic stem cell ,Chromatin ,Cell biology ,DNA-Binding Proteins ,KLF4 ,ZIC3 ,Urine cells ,Stem cell ,Transcription Factors - Abstract
Background Four transcription factors, Oct4, Sox2, Klf4, and c-Myc (the Yamanka factors), can reprogram somatic cells to induced pluripotent stem cells (iPSCs). Many studies have provided a number of alternative combinations to the non-Yamanaka factors. However, it is clear that many additional transcription factors that can generate iPSCs remain to be discovered. Methods The chromatin accessibility and transcriptional level of human embryonic stem cells and human urine cells were compared by Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq) and RNA sequencing (RNA-seq) to identify potential reprogramming factors. Selected transcription factors were employed to reprogram urine cells, and the reprogramming efficiency was measured. Urine-derived iPSCs were detected for pluripotency by Immunofluorescence, quantitative polymerase chain reaction, RNA sequencing and teratoma formation test. Finally, we assessed the differentiation potential of the new iPSCs to cardiomyocytes in vitro. Results ATAC-seq and RNA-seq datasets predicted TEAD2, TEAD4 and ZIC3 as potential factors involved in urine cell reprogramming. Transfection of TEAD2, TEAD4 and ZIC3 (in the presence of Yamanaka factors) significantly improved the reprogramming efficiency of urine cells. We confirmed that the newly generated iPSCs possessed pluripotency characteristics similar to normal H1 embryonic stem cells. We also confirmed that the new iPSCs could differentiate to functional cardiomyocytes. Conclusions In conclusion, TEAD2, TEAD4 and ZIC3 can increase the efficiency of reprogramming human urine cells into iPSCs, and provides a new stem cell sources for the clinical application and modeling of cardiovascular disease. Graphical abstract more...
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- 2021
17. MuRF1 deficiency prevents age‐related fat weight gain, possibly through accumulation of PDK4 in skeletal muscle mitochondria in older mice
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Siegfried Labeit, Motoko Oarada, Reiko Nakao, Iori Sakakibara, Anayt Ulla, Tasuku Maeda, Koichi Sairyo, Kosuke Sugiura, Takayuki Uchida, Koji Kishimoto, Katsuya Hirasaka, and Takeshi Nikawa
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medicine.medical_specialty ,Ubiquitin-Protein Ligases ,Muscle Proteins ,Adipose tissue ,PDK4 ,Mitochondrion ,Weight Gain ,Tripartite Motif Proteins ,Mice ,Atrophy ,Internal medicine ,medicine ,Animals ,Orthopedics and Sports Medicine ,Muscle, Skeletal ,biology ,Chemistry ,Pyruvate Dehydrogenase Acetyl-Transferring Kinase ,Skeletal muscle ,Lipid metabolism ,medicine.disease ,Muscle atrophy ,Mitochondria ,Mitochondria, Muscle ,Ubiquitin ligase ,Muscular Atrophy ,Endocrinology ,medicine.anatomical_structure ,Adipose Tissue ,biology.protein ,medicine.symptom ,Protein Kinases - Abstract
Recent studies show that muscle mass and metabolic function are interlinked. Muscle RING finger 1 (MuRF1) is a critical muscle-specific ubiquitin ligase associated with muscle atrophy. Yet, the molecular target of MuRF1 in atrophy and aging remains unclear. We examined the role of MuRF1 in aging, using MuRF1-deficient (MuRF1-/- ) mice in vivo, and MuRF1-overexpressing cell in vitro. MuRF1 deficiency partially prevents age-induced skeletal muscle loss in mice. Interestingly, body weight and fat mass of >7-month-old MuRF1-/- mice were lower than in MuRF1+/+ mice. Serum and muscle metabolic parameters and results of indirect calorimetry suggest significantly higher energy expenditure and enhanced lipid metabolism in 3-month-old MuRF1-/- mice than in MuRF1+/+ mice, resulting in suppressed adipose tissue gain during aging. Pyruvate dehydrogenase kinase 4 (PDK4) is crucial for a switch from glucose to lipid metabolism, and the interaction between MuRF1 and PDK4 was examined. PDK4 protein levels were elevated in mitochondria from the skeletal muscle in MuRF1-/- mice. In vitro, MuRF1 interacted with PDK4 but did not induce degradation through ubiquitination. Instead, SUMOylation of PDK4 was detected in MuRF1-overexpressing cells, in contrast to cells without the RING domain of MuRF1. MuRF1 deficiency enhances lipid metabolism possibly by upregulating PDK4 localization into mitochondrial through prevention of SUMOylation. Inhibition of MuRF1-mediated PDK4 SUMOylation is a potential therapeutic target for age-related dysfunction of lipid metabolism and muscle atrophy. This article is protected by copyright. All rights reserved. more...
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- 2021
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18. Mutation in FBXO32 causes dilated cardiomyopathy through up-regulation of ER-stress mediated apoptosis
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Kunhi Muhammed, Abdullah M. Assiri, Dilek Colak, Salma Awad Mahmoud, Maya Hammonds, Jehad Al-Buraiki, Waleed AlHabeeb, Olfat Al-Harazi, Coralie Poizat, and Nadya Al-Yacoub
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0301 basic medicine ,Cardiomyopathy, Dilated ,Programmed cell death ,QH301-705.5 ,Cardiomyopathy ,Mutation, Missense ,Cardiology ,Medicine (miscellaneous) ,Muscle Proteins ,Apoptosis ,CHOP ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Genetics ,Biology (General) ,Transcription factor ,FBXO32 ,SKP Cullin F-Box Protein Ligases ,Endoplasmic reticulum ,Autophagy ,medicine.disease ,Endoplasmic Reticulum Stress ,Cell biology ,Up-Regulation ,Gene regulation ,030104 developmental biology ,030220 oncology & carcinogenesis ,Unfolded protein response ,biological phenomena, cell phenomena, and immunity ,General Agricultural and Biological Sciences - Abstract
Endoplasmic reticulum (ER) stress induction of cell death is implicated in cardiovascular diseases. Sustained activation of ER-stress induces the unfolded protein response (UPR) pathways, which in turn activate three major effector proteins. We previously reported a missense homozygous mutation in FBXO32 (MAFbx, Atrogin-1) causing advanced heart failure by impairing autophagy. In the present study, we performed transcriptional profiling and biochemical assays, which unexpectedly revealed a reduced activation of UPR effectors in patient mutant hearts, while a strong up-regulation of the CHOP transcription factor and of its target genes are observed. Expression of mutant FBXO32 in cells is sufficient to induce CHOP-associated apoptosis, to increase the ATF2 transcription factor and to impair ATF2 ubiquitination. ATF2 protein interacts with FBXO32 in the human heart and its expression is especially high in FBXO32 mutant hearts. These findings provide a new underlying mechanism for FBXO32-mediated cardiomyopathy, implicating abnormal activation of CHOP. These results suggest alternative non-canonical pathways of CHOP activation that could be considered to develop new therapeutic targets for the treatment of FBXO32-associated DCM., Al-Yacoub et al. investigate the consequences of FBXO32 mutation on dilated cardiomyopathy. ER stress, abnormal CHOP activation and CHOP-induced apoptosis with no UPR effector activation are found to underlie the FBXO32 mutation induced cardiomyopathy, suggesting an alternative pathway that can be considered to develop new therapeutic targets for its treatment. more...
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- 2021
19. Resistance Exercise, Aging, Disuse, and Muscle Protein Metabolism
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Stuart M. Phillips, James McKendry, Tanner Stokes, and Jonathan C Mcleod
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Aging ,medicine.medical_specialty ,Anabolism ,Glucose uptake ,Resistance training ,Muscle Proteins ,Skeletal muscle ,Resistance Training ,Endogeny ,Metabolism ,Biology ,medicine.disease ,Endocrinology ,medicine.anatomical_structure ,Internal medicine ,Sarcopenia ,Basal metabolic rate ,medicine ,Humans ,Muscle, Skeletal ,Exercise - Abstract
Skeletal muscle is the organ of locomotion, its optimal function is critical for athletic performance, and is also important for health due to its contribution to resting metabolic rate and as a site for glucose uptake and storage. Numerous endogenous and exogenous factors influence muscle mass. Much of what is currently known regarding muscle protein turnover is owed to the development and use of stable isotope tracers. Skeletal muscle mass is determined by the meal- and contraction-induced alterations of muscle protein synthesis and muscle protein breakdown. Increased loading as resistance training is the most potent nonpharmacological strategy by which skeletal muscle mass can be increased. Conversely, aging (sarcopenia) and muscle disuse lead to the development of anabolic resistance and contribute to the loss of skeletal muscle mass. Nascent omics-based technologies have significantly improved our understanding surrounding the regulation of skeletal muscle mass at the gene, transcript, and protein levels. Despite significant advances surrounding the mechanistic intricacies that underpin changes in skeletal muscle mass, these processes are complex, and more work is certainly needed. In this article, we provide an overview of the importance of skeletal muscle, describe the influence that resistance training, aging, and disuse exert on muscle protein turnover and the molecular regulatory processes that contribute to changes in muscle protein abundance. © 2021 American Physiological Society. Compr Physiol 11:2249-2278, 2021. more...
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- 2021
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20. Establishment and characterization of NCC-ssRMS2-C1: a novel patient-derived cell line of spindle cell/sclerosing rhabdomyosarcoma
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Jun Sugaya, Akihiko Yoshida, Fumihiko Nakatani, Tadashi Kondo, Yooksil Sin, Rei Noguchi, Takuya Ono, Yuki Yoshimatsu, Fumitaka Takeshita, Ryuto Tsuchiya, Seiji Ohtori, Akane Sei, and Akira Kawai
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Adult ,Male ,0301 basic medicine ,Cancer Research ,Cell ,Muscle Proteins ,Antineoplastic Agents ,Biology ,Sclerosing rhabdomyosarcoma ,Romidepsin ,Bortezomib ,Nuclear Receptor Coactivator 2 ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Depsipeptides ,Rhabdomyosarcoma ,medicine ,Humans ,MyoD Protein ,Sarcoma ,Cell Biology ,medicine.disease ,Spindle Cell/Sclerosing Rhabdomyosarcoma ,030104 developmental biology ,medicine.anatomical_structure ,Head and Neck Neoplasms ,030220 oncology & carcinogenesis ,Mutation ,Dactinomycin ,Cancer research ,Gene Fusion ,Stem cell ,Trabectedin ,Transcription Factors ,medicine.drug - Abstract
Spindle cell/sclerosing rhabdomyosarcoma (ssRMS) is a rare subtype of rhabdomyosarcoma (RMS) that has fascicular spindle cell and/or sclerosing morphology. SsRMS has a diverse molecular background and is categorized into three groups: congenital/infantile ssRMS with a gene fusion involving the NCOA2 and VGLL2, ssRMS with the MYOD1 mutation, and ssRMS with no recurrent identifiable genetic alterations. Because ssRMS is a newly defined disease concept of RMS, the optimal treatment methods have not been determined. This results in unfavorable prognosis and consequently signals the urgent need for continuous research. Patient-derived cell lines are essential tools in basic and translational research. However, only two ssRMS cell lines with the MYOD1 mutation have been reported to date. Thus, we established a novel ssRMS cell line named NCC-ssRMS2-C1 using a surgically resected tumor tissue from an adult ssRMS patient. NCC-ssRMS2-C1 cells retained the copy number alterations corresponding to the original tumor and are categorized into the group with no recurrent identifiable genetic alterations. NCC-ssRMS2-C1 cells demonstrated constant proliferation, spheroid formation, and capability for invasion in vitro, reflecting the malignant features of the original tumor tissue. In a drug screening test, ssRMS demonstrated remarkable sensitivity to romidepsin, trabectedin, actinomycin D, and bortezomib. Hence, we conclude that the NCC-ssRMS2-C1 cell line is the first ssRMS cell line which belongs to the group with no recurrent identifiable genetic alterations, and it will be a useful resource in both basic and translational studies for ssRMS. more...
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- 2021
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21. Kindlin-2 in Sertoli cells is essential for testis development and male fertility in mice
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Hongquan Zhang, Bing Li, Jun Zhan, Xiaochun Chi, Miao Yu, Zhenbin Wang, Weiwei Luo, Tiantian Su, Cheng Liu, Jiagui Song, Hui-ying He, Tianzhuo Wang, and Zhen Li
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Male ,Cancer Research ,Male factor infertility ,Phagocytosis ,Immunology ,Muscle Proteins ,Apoptosis ,Mice, Transgenic ,Biology ,Article ,Mice ,Cellular and Molecular Neuroscience ,Testis ,medicine ,Animals ,Humans ,Cells, Cultured ,Infertility, Male ,Hippo signaling pathway ,Sertoli Cells ,QH573-671 ,Nucleus localization ,Cartilage ,Cell Biology ,Sertoli cell ,Sperm ,Cell biology ,Cytoskeletal Proteins ,Fertility ,HEK293 Cells ,medicine.anatomical_structure ,Phosphorylation ,Cytology - Abstract
Kindlin-2 is known to play important roles in the development of mesoderm-derived tissues including myocardium, smooth muscle, cartilage and blood vessels. However, nothing is known for the role of Kindlin-2 in mesoderm-derived reproductive organs. Here, we report that loss of Kindlin-2 in Sertoli cells caused severe testis hypoplasia, abnormal germ cell development and complete infertility in male mice. Functionally, loss of Kindlin-2 inhibits proliferation, increases apoptosis, impairs phagocytosis in Sertoli cells and destroyed the integration of blood-testis barrier structure in testes. Mechanistically, Kindlin-2 interacts with LATS1 and YAP, the key components of Hippo pathway. Kindlin-2 impedes LATS1 interaction with YAP, and depletion of Kindlin-2 enhances LATS1 interaction with YAP, increases YAP phosphorylation and decreases its nuclear translocation. For clinical relevance, lower Kindlin-2 expression and decreased nucleus localization of YAP was found in SCOS patients. Collectively, we demonstrated that Kindlin-2 in Sertoli cells is essential for sperm development and male reproduction. more...
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- 2021
22. Glutamine supplementation stimulates cell proliferation in skeletal muscle and cultivated myogenic cells of low birth weight piglets
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Zeyang Li, Q. Sciascia, Johannes Schregel, Elke Albrecht, Yaolu Zhao, Steffen Maak, Katja Stange, and Cornelia C. Metges
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0301 basic medicine ,Muscle tissue ,DNA Replication ,Male ,Cell type ,Satellite Cells, Skeletal Muscle ,Swine ,Glutamine ,Science ,Muscle Proteins ,Biology ,Article ,Muscle hypertrophy ,Andrology ,03 medical and health sciences ,Cell growth ,Animal physiology ,Muscle stem cells ,medicine ,Animals ,Birth Weight ,RNA, Messenger ,Muscle, Skeletal ,Cells, Cultured ,Growth Disorders ,Swine Diseases ,Multidisciplinary ,Alanine ,030102 biochemistry & molecular biology ,Dose-Response Relationship, Drug ,Myogenesis ,Skeletal muscle ,Gene Expression Regulation, Developmental ,Immunohistochemistry ,Animals, Suckling ,Culture Media ,030104 developmental biology ,medicine.anatomical_structure ,Bromodeoxyuridine ,Cell culture ,Dietary Supplements ,Medicine ,Transcription ,Cell Division - Abstract
Muscle growth of low birth weight (LBW) piglets may be improved with adapted nutrition. This study elucidated effects of glutamine (Gln) supplementation on the cellular muscle development of LBW and normal birth weight (NBW) piglets. Male piglets (n = 144) were either supplemented with 1 g Gln/kg body weight or an isonitrogeneous amount of alanine (Ala) between postnatal day 1 and 12 (dpn). Twelve piglets per group were slaughtered at 5, 12 and 26 dpn, one hour after injection with Bromodeoxyuridine (BrdU, 12 mg/kg). Muscle samples were collected and myogenic cells were isolated and cultivated. Expression of muscle growth related genes was quantified with qPCR. Proliferating, BrdU-positive cells in muscle sections were detected with immunohistochemistry indicating different cell types and decreasing proliferation with age. More proliferation was observed in muscle tissue of LBW-GLN than LBW-ALA piglets at 5 dpn, but there was no clear effect of supplementation on related gene expression. Cell culture experiments indicated that Gln could promote cell proliferation in a dose dependent manner, but expression of myogenesis regulatory genes was not altered. Overall, Gln supplementation stimulated cell proliferation in muscle tissue and in vitro in myogenic cell culture, whereas muscle growth regulatory genes were barely altered. more...
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- 2021
23. Myospryn deficiency leads to impaired cardiac structure and function and schizophrenia-associated symptoms
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Aimilia Varela, Ioanna Kostavasili, Alexia Polissidis, Ismini Kloukina, Manolis Mavroidis, Modestos Nakos-Bimpos, Mary Tsikitis, Elsa Tsoupri, Yassemi Capetanaki, Despoina Miliou, Constantinos H. Davos, and Eleni Vasilaki more...
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Male ,0301 basic medicine ,Histology ,Cardiomyopathy ,Muscle Proteins ,Biology ,Mitochondrion ,Pathology and Forensic Medicine ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Monoaminergic ,medicine ,Animals ,Humans ,Sarcomere organization ,Prepulse inhibition ,Myocardium ,Intracellular Signaling Peptides and Proteins ,Cell Biology ,medicine.disease ,Phenotype ,Cell biology ,Dysbindin ,030104 developmental biology ,Schizophrenia ,Female ,Desmin ,030217 neurology & neurosurgery - Abstract
The desmin-associated protein myospryn, encoded by the cardiomyopathy-associated gene 5 (CMYA5), is a TRIM-like protein associated to the BLOC-1 (Biogenesis of Lysosomes Related Organelles Complex 1) protein dysbindin. Human myospryn mutations are linked to both cardiomyopathy and schizophrenia; however, there is no evidence of a direct causative link of myospryn to these diseases. Therefore, we sought to unveil the role of myospryn in heart and brain. We have genetically inactivated the myospryn gene by homologous recombination and demonstrated that myospryn null hearts have dilated phenotype and compromised cardiac function. Ultrastructural analyses revealed that the sarcomere organization is not obviously affected; however, intercalated disk (ID) integrity is impaired, along with mislocalization of ID and sarcoplasmic reticulum (SR) protein components. Importantly, cardiac and skeletal muscles of myospryn null mice have severe mitochondrial defects with abnormal internal vacuoles and extensive cristolysis. In addition, swollen SR and T-tubules often accompany the mitochondrial defects, strongly implying a potential link of myospryn together with desmin to SR- mitochondrial physical and functional cross-talk. Furthermore, given the reported link of human myospryn mutations to schizophrenia, we performed behavioral studies, which demonstrated that myospryn-deficient male mice display disrupted startle reactivity and prepulse inhibition, asocial behavior, decreased exploratory behavior, and anhedonia. Brain neurochemical and ultrastructural analyses revealed prefrontal-striatal monoaminergic neurotransmitter defects and ultrastructural degenerative aberrations in cerebellar cytoarchitecture, respectively, in myospryn-deficient mice. In conclusion, myospryn is essential for both cardiac and brain structure and function and its deficiency leads to cardiomyopathy and schizophrenia-associated symptoms. more...
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- 2021
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24. Expanding the clinical spectrum and management of Traboulsi syndrome: report on two siblings homozygous for a novel pathogenic variant in ASPH
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Tom Van Hoorde, Bart P. Leroy, Irina Balikova, Mattias Van Heetvelde, Hannah Verdin, Elfride De Baere, Dimitri Roels, Philippe Huyghe, Elke O. Kreps, and Fanny Nerinckx
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Male ,medicine.medical_specialty ,Adolescent ,genetic structures ,media_common.quotation_subject ,Nonsense ,Visual Acuity ,Iris ,Muscle Proteins ,Cataract Extraction ,Consanguinity ,Slit Lamp Microscopy ,Ectopia Lentis ,Mixed Function Oxygenases ,Frameshift mutation ,Craniofacial Abnormalities ,Young Adult ,Exon ,Ophthalmology ,Exome Sequencing ,medicine ,Humans ,Ectopia lentis ,Genetics (clinical) ,Genetic testing ,media_common ,medicine.diagnostic_test ,biology ,business.industry ,Siblings ,Calcium-Binding Proteins ,Membrane Proteins ,Exons ,medicine.disease ,eye diseases ,ASPH ,Codon, Nonsense ,Pediatrics, Perinatology and Child Health ,biology.protein ,Female ,sense organs ,Bleb (medicine) ,business - Abstract
Background Traboulsi syndrome is a very rare, syndromic form of ectopia lentis that is potentially sight-threatening at a young age. It is characterized by typical facial, skeletal and ocular signs. Materials and methods Two siblings, born to consanguineous parents, with a clinical phenotype consistent with Traboulsi syndrome, underwent extensive ophthalmic imaging and exome-based genetic testing. Both were treated with unilateral clear lens extraction via a limbal approach. Results Two siblings, one male and one female, presented with systemic and ocular features consistent with Traboulsi syndrome. Lens subluxation was present in all 4fouraffected eyes, and spontaneous subconjunctival bleb formation was detected in one eye. This eye also showed evidence of keratoconus-related corneal thinning. The clinical diagnosis of Traboulsi syndrome was confirmed molecularly. A homozygous, novel, pathogenic nonsense variant was identified in exon 25 of the ASPH gene: c.2181_2183dup, p.(Val727_Trp728insTer). Excellent visual outcomes following clear lens extraction and postoperative rigid gas-permeable contact lens fitting were obtained. Conclusions We expanded the genetic spectrum of Traboulsi syndrome with a novel frameshift variant in the ASPH gene. We showed that lensectomy followed by gas-permeable contact lenses is an efficient therapeutic approach to treat lens subluxation in Traboulsi syndrome. However, lifelong follow-up is crucial to avoid (late) postoperative complications. more...
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- 2021
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25. A novel tumor suppressor role of myosin light chain kinase splice variants through downregulation of the TEAD4/CD44 axis
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Jerrold R. Turner, Tsung-Chun Lee, Yu Chen Pai, Linda C.H. Yu, Yen Ju Huang, and Been-Ren Lin
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Male ,0301 basic medicine ,Cancer Research ,Myosin light-chain kinase ,Carcinogenesis ,Muscle Proteins ,Apoptosis ,macromolecular substances ,medicine.disease_cause ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Cell Movement ,Biomarkers, Tumor ,Tumor Cells, Cultured ,medicine ,Animals ,Humans ,Phosphorylation ,Myosin-Light-Chain Kinase ,Transcription factor ,Adaptor Proteins, Signal Transducing ,Cell Proliferation ,Mice, Knockout ,YAP1 ,Hippo signaling pathway ,biology ,Chemistry ,Cell Cycle ,CD44 ,TEA Domain Transcription Factors ,YAP-Signaling Proteins ,MYLK ,General Medicine ,Cell cycle ,Prognosis ,Cell biology ,DNA-Binding Proteins ,Gene Expression Regulation, Neoplastic ,Mice, Inbred C57BL ,Survival Rate ,Alternative Splicing ,Hyaluronan Receptors ,030104 developmental biology ,030220 oncology & carcinogenesis ,Colonic Neoplasms ,biology.protein ,Transcription Factors - Abstract
Myosin light chain kinase (MLCK) regulates actinomyosin contraction. Two splice variants of long MLCK are expressed in epithelial cells and divergently regulate gut barrier functions; reduced MLCK levels in human colorectal cancers (CRC) with unclarified significance have been reported. CRC are solid tumors clonally sustained by stem cells highly expressing CD44 and CD133. The aim was to investigate the role of MLCK splice variants in CRC tumorigenesis. We found lower MLCK1/2 and higher CD44 expression in human CRC, but no change in CD133 or LGR5. Large-scale bioinformatics showed an inverse relationship between MYLK and CD44 in human sample gene datasets. A 3-fold increased tumor burden was observed in MLCK(−/−) mice compared with wild-type (WT) mice in a chemical-induced CRC model. Primary tumorspheres derived from the MLCK(−/−) mice displayed larger sizes and higher CD44 transcript levels than those from the WT mice. Bioinformatics revealed binding of TEAD4 (a transcriptional enhancer factor family member in the Hippo pathway) to CD44 promoter, which was confirmed by luciferase reporter assay. Individually expressing MLCK1 and MLCK2 variants in the MLCK-knockout (KO) Caco-2 cells inhibited the nuclear localization of TEAD4 cofactors, VGLL3 and YAP1, respectively, and both variants reduced the CD44 transcription. Accelerated cell cycle transit was observed in the MLCK-KO cells, whereby expression of MLCK1/2 variants counterbalanced the cell hyperproliferation. In conclusion, MLCK1/2 variants are novel tumor suppressors by downregulating the TEAD4/CD44 axis via reducing nuclear translocation of distinct transcriptional coactivators. The reduction of epithelial MLCKs, especially isoform 2, may drive cancer stemness and tumorigenesis. more...
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- 2021
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26. Uncovering potential novel biomarkers and immune infiltration characteristics in persistent atrial fibrillation using integrated bioinformatics analysis
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Ting Yin, Yue Hu, Ailin Liu, Yufei Zhou, Hong Zhu, Jie Liu, Qi Wu, Shengjue Xiao, and Defeng Pan
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differentially expressed genes ,pathways ,Muscle Proteins ,02 engineering and technology ,Biology ,Pathogenesis ,Immune system ,0502 economics and business ,microRNA ,Atrial Fibrillation ,0202 electrical engineering, electronic engineering, information engineering ,QA1-939 ,Humans ,Gene Regulatory Networks ,Gene ,Microarray analysis techniques ,immune infiltration ,Applied Mathematics ,Calcineurin ,05 social sciences ,Computational Biology ,General Medicine ,persistent atrial fibrillation ,DNA-Binding Proteins ,Computational Mathematics ,MicroRNAs ,microrna-transcription factor-mrna network ,Gene Expression Regulation ,Modeling and Simulation ,Cancer research ,020201 artificial intelligence & image processing ,Signal transduction ,General Agricultural and Biological Sciences ,050203 business & management ,CD8 ,Biomarkers ,TP248.13-248.65 ,Mathematics ,Biotechnology - Abstract
Atrial fibrillation (AF) is the most common cardiac arrhythmia. This study aimed to identify potential novel biomarkers for persistent AF (pAF) using integrated analyses and explore the immune cell infiltration in this pathological process. Three pAF datasets (GSE31821, GSE41177, and GSE79768) from the Gene Expression Omnibus (GEO) database were integrated with the elimination of batch effects. 264 differentially expressed genes (DEGs) were identified using Linear models for microarray data (LIMMA), 12 modules were screened out by weighted gene co-expression network analysis (WGCNA) in pAF compared with normal controls. Subsequently, common genes (CGs) were identified as the intersection of DEGs and genes in the most significant module. Functional enrichment analysis showed that CGs were mainly enriched in the "Calcineurin-NFAT (nuclear factor of activated T-cells)" signaling pathway, particularly regulator of calcineurin 1 (RCAN1), and protein phosphatase 3 regulatory subunit B, alpha (PPP3R1). Ulteriorly, the microRNA-transcription factor-mRNA network revealed that microRNA-34a-5p could target both RCAN1 and PPP3R1 in the pAF pathogenesis. Finally, immune infiltration analysis by CIBERSORT, a versatile computational method, displayed a higher level of monocytes, dendritic cells and neutrophils, as well as a lower level of CD8+ T cells and T cells regulatory (Tregs) in pAF compared with the control group. In conclusion, our present study revealed several novel pAF-associated genes, miRNAs, and pathways, including microRNA-34a-5p, which might target RCAN1 and PPP3R1 to regulate pAF through the calcineurin-NFAT signaling pathway. In addition, there was a difference in immune infiltration between patients with pAF and normal groups and immune cells might interact with specific genes in pAF. more...
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- 2021
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27. Molecular motion and tridimensional nanoscale localization of kindlin control integrin activation in focal adhesions
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Birgit Kastberger, Adrien Joly, Ralph T. Böttcher, Reinhard Fässler, Zeynep Karatas, Bernhard Wehrle-Haller, Thomas Orré, Jean-Baptiste Sibarita, Clément Cabriel, Olivier Rossier, Sandrine Lévêque-Fort, Grégory Giannone, Interdisciplinary Institute for Neuroscience (IINS), Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS), Institut des Sciences Moléculaires d'Orsay (ISMO), and Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS) more...
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0301 basic medicine ,Integrins ,[SDV]Life Sciences [q-bio] ,General Physics and Astronomy ,Muscle Proteins ,Plasma protein binding ,Mice ,0302 clinical medicine ,Cells, Cultured ,Mice, Knockout ,[PHYS]Physics [physics] ,Cultured ,Multidisciplinary ,biology ,Chemistry ,Membrane Proteins/genetics/metabolism ,Cell biology ,Neoplasm Proteins ,Pleckstrin homology domain ,Cytoskeletal Proteins/genetics/metabolism ,Protein Binding ,Muscle Proteins/genetics/metabolism ,Cells ,Knockout ,Science ,Integrin ,Biophysics ,Neoplasm Proteins/genetics/metabolism ,General Biochemistry, Genetics and Molecular Biology ,Article ,Focal adhesion ,03 medical and health sciences ,Motion ,Cell Adhesion ,Animals ,Humans ,ddc:612 ,Cell adhesion ,Focal Adhesions ,Binding Sites ,Cell Membrane/metabolism ,Focal Adhesions/metabolism ,Cell Membrane ,Membrane Proteins ,General Chemistry ,Actin cytoskeleton ,Fibronectin ,Cytoskeletal Proteins ,030104 developmental biology ,Membrane protein ,Mutation ,Integrins/genetics/metabolism ,biology.protein ,030217 neurology & neurosurgery - Abstract
Focal adhesions (FAs) initiate chemical and mechanical signals involved in cell polarity, migration, proliferation and differentiation. Super-resolution microscopy revealed that FAs are organized at the nanoscale into functional layers from the lower plasma membrane to the upper actin cytoskeleton. Yet, how FAs proteins are guided into specific nano-layers to promote interaction with given targets is unknown. Using single protein tracking, super-resolution microscopy and functional assays, we link the molecular behavior and 3D nanoscale localization of kindlin with its function in integrin activation inside FAs. We show that immobilization of integrins in FAs depends on interaction with kindlin. Unlike talin, kindlin displays free diffusion along the plasma membrane outside and inside FAs. We demonstrate that the kindlin Pleckstrin Homology domain promotes membrane diffusion and localization to the membrane-proximal integrin nano-layer, necessary for kindlin enrichment and function in FAs. Using kindlin-deficient cells, we show that kindlin membrane localization and diffusion are crucial for integrin activation, cell spreading and FAs formation. Thus, kindlin uses a different route than talin to reach and activate integrins, providing a possible molecular basis for their complementarity during integrin activation., Focal adhesions (FAs) initiate chemical and mechanical signals involved in cell polarity, migration, proliferation and differentiation. Here, authors combine single protein tracking, super-resolution microscopy and functional assays, which allow correlating the molecular behaviour and 3D nanoscale localization of kindlin with its function in integrin activation inside FAs. more...
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- 2021
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28. Ethylmalonic encephalopathy ETHE1 p. D165H mutation alters the mitochondrial function in human skeletal muscle proteome
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Bindu Parayil Sankaran, Periyasamy Govindaraj, Tripti Khanna, Madhu Nagappa, Arun B Taly, Sekar Deepha, Gajanan Sathe, Akhilesh Pandey, and Narayanappa Gayathri
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Adult ,Male ,Proteomics ,0301 basic medicine ,Nucleocytoplasmic Transport Proteins ,Proteome ,Quantitative proteomics ,Down-Regulation ,Muscle Proteins ,Oxidative phosphorylation ,Mitochondrion ,Biology ,Oxidative Phosphorylation ,Mitochondrial Proteins ,03 medical and health sciences ,0302 clinical medicine ,Ethylmalonic encephalopathy ,medicine ,Humans ,Muscle, Skeletal ,Molecular Biology ,Purpura ,Brain Diseases, Metabolic, Inborn ,Skeletal muscle ,Cell Biology ,medicine.disease ,Mitochondria, Muscle ,030104 developmental biology ,medicine.anatomical_structure ,Biochemistry ,Inborn error of metabolism ,Mutation ,Molecular Medicine ,ETHE1 ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
Ethylmalonic encephalopathy (EE) is a rare autosomal recessive inborn error of metabolism. To study the molecular effects of ETHE1 p. D165H mutation, we employed mass spectrometry-based mitochondrial proteome and phosphoproteome profiling in the human skeletal muscle. Eighty-six differentially altered proteins were identified, of which thirty-seven mitochondrial proteins were differentially expressed, and most of the proteins (37%) were down-regulated in the OXPHOS complex-IV. Also, nine phosphopeptides that correspond to eight mitochondrial proteins were significantly affected in EE patient. These altered proteins recognized are involved in several pathways and molecular functions, predominantly in oxidoreductase activity. This is the first study that has integrated proteome and phosphoproteome of skeletal muscle and identified multiple proteins associated in the pathogenesis of EE. more...
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- 2021
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29. Effect of flavonoids from Lycium barbarum leaves on the oxidation of myofibrillar proteins in minced mutton during chilled storage
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Tingting Kou, Yinhong Niu, Jinghua Chen, and Yanli Fan
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China ,Antioxidant ,030309 nutrition & dietetics ,Rutin ,medicine.medical_treatment ,Radical ,Flavonoid ,Muscle Proteins ,Protein oxidation ,High-performance liquid chromatography ,Antioxidants ,03 medical and health sciences ,chemistry.chemical_compound ,0404 agricultural biotechnology ,Myofibrils ,medicine ,Animals ,Food science ,Flavonoids ,chemistry.chemical_classification ,0303 health sciences ,Sheep ,biology ,Chemistry ,Tryptophan ,food and beverages ,04 agricultural and veterinary sciences ,Lycium ,biology.organism_classification ,040401 food science ,Cold Temperature ,Meat Products ,Plant Leaves ,Red Meat ,Food Storage ,Oxidation-Reduction ,Food Science - Abstract
Herein, we report the effect of flavonoids from Lycium barbarum leaves (LBLF) on myofibrillar proteins (MP) in minced mutton during chilled storage (4 ± 1 ℃). High performance liquid chromatography (HPLC) analysis showed that the total flavonoid content in LBLF was 322.0 mg/g, of which the rutin content was 297.6 mg/g. The effect of 0.5%, 1.0%, and 1.5% LBLF on the structure and thermodynamic properties of MP in minced mutton was studied systematically. Tyrosine and tryptophan of MP samples treated with LBLF were converted from an exposed state to an embedded state. The interaction between LBLF and MP quenched the internal fluorescence, and improved the thermal stability of MP. The addition of LBLF significantly reduced the carbonyl and sulfhydryl contents of MP (p < 0.05), and decreased the surface hydrophobicity of MP in a dose-dependent manner. Our results indicate that LBLF can combine with free radicals produced by protein oxidation, block the free radical oxidation chain reaction, and inhibit the oxidation of MP. Therefore, LBLF may have great potential as a natural antioxidant in meats and meat products during chilled storage. PRACTICAL APPLICATION: Lycium barbarum is widely distributed in China, especially in Qinghai and Ningxia. The results of this study suggest that flavonoids extracted from L. barbarum leaves may be an effective natural antioxidant for the preservation of meats and meat products. more...
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- 2021
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30. A new degree of complexi(n)ty in the regulation of GLUT4 trafficking
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Sandrine Horman, Marine De Loof, Christophe Beauloye, Luc Bertrand, Laurent Bultot, and UCL - SSS/IREC/CARD - Pôle de recherche cardiovasculaire
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Monosaccharide Transport Proteins ,medicine.medical_treatment ,Glucose uptake ,muscle metabolism ,Muscle Proteins ,030209 endocrinology & metabolism ,Stimulation ,Type 2 diabetes ,insulin signalling ,Biology ,Biochemistry ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Humans ,Insulin ,Myocyte ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Cell Membrane ,Glucose transporter ,glucose transport ,Cell Biology ,Metabolism ,medicine.disease ,Cell biology ,Glucose ,Diabetes Mellitus, Type 2 ,biology.protein ,complexin ,GLUT4 - Abstract
Loss of the insulin-stimulated glucose uptake in muscle is a crucial event participating in the defect of whole-body metabolism in type 2 diabetes. Therefore, identification by Pavarotti et al. (Biochem. J (2021) 478 (2): 407–422) of complexin-2 as an important contributor to glucose transporter 4 (GLUT4) translocation to muscle cell plasma membrane upon insulin stimulation is essential. The present commentary discusses the biological importance of the findings and proposes future challenges and opportunities. more...
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- 2021
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31. Marginally reduced maternal hepatic and splenic ferroportin under severe nutritional iron deficiency in pregnancy maintains systemic iron supply
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Aneta Jończy, Aleksandra Bednarz, Rafał Mazgaj, Eunice Sindhuvi Edison, Olga Haberkiewicz, Rafał R. Starzyński, Robert Staroń, Małgorzata Lenartowicz, Ewa Smuda, and Paweł Lipiński
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Erythrocyte Indices ,Placenta ,Ferroportin ,Muscle Proteins ,Hemoglobins ,Mice ,0302 clinical medicine ,Pregnancy ,Cation Transport Proteins ,Maternal-Fetal Exchange ,Research Articles ,reproductive and urinary physiology ,biology ,Erythrocyte Aging ,Iron Deficiencies ,Hematology ,Iron deficiency ,Up-Regulation ,medicine.anatomical_structure ,Liver ,Organ Specificity ,030220 oncology & carcinogenesis ,embryonic structures ,Cytokines ,Erythropoiesis ,Female ,Iron, Dietary ,Research Article ,medicine.medical_specialty ,Mice, 129 Strain ,Duodenum ,Anemia ,Iron ,Nerve Tissue Proteins ,03 medical and health sciences ,Fetus ,Hepcidins ,Phagocytosis ,Hepcidin ,Internal medicine ,medicine ,Animals ,Macrophages ,Membrane Proteins ,medicine.disease ,Pregnancy Complications ,Endocrinology ,biology.protein ,Carrier Proteins ,Spleen ,030215 immunology - Abstract
The demand for iron is high in pregnancy to meet the increased requirements for erythropoiesis. Even pregnant females with initially iron‐replete stores develop iron‐deficiency anemia, due to inadequate iron absorption. In anemic females, the maternal iron supply is dedicated to maintaining iron metabolism in the fetus and placenta. Here, using a mouse model of iron deficiency in pregnancy, we show that iron recycled from senescent erythrocytes becomes a predominant source of this microelement that can be transferred to the placenta in females with depleted iron stores. Ferroportin is a key protein in the molecular machinery of cellular iron egress. We demonstrate that under iron deficiency in pregnancy, levels of ferroportin are greatly reduced in the duodenum, placenta and fetal liver, but not in maternal liver macrophages and in the spleen. Although low expression of both maternal and fetal hepcidin predicted ferroportin up‐regulation in examined locations, its final expression level was very likely correlated with tissue iron status. Our results argue that iron released into the circulation of anemic females is taken up by the placenta, as evidenced by high expression of iron importers on syncytiotrophoblasts. Then, a substantial decrease in levels of ferroportin on the basolateral side of syncytiotrophoblasts, may be responsible for the reduced transfer of iron to the fetus. As attested by the lowest decrease in iron content among analyzed tissues, some part is retained in the placenta. These findings confirm the key role played by ferroportin in tuning iron turnover in iron‐deficient pregnant mouse females and their fetuses. more...
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- 2021
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32. TAGLN Is Downregulated by TRAF6-Mediated Proteasomal Degradation in Prostate Cancer Cells
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Gaozhen Jia, Fuping Wen, Xiaochen Sun, Chenghua Yang, Haolan Yu, Chenxia Sun, Zhenyang Dong, and Wei Bao
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Male ,0301 basic medicine ,Proteasome Endopeptidase Complex ,Cancer Research ,Down-Regulation ,Muscle Proteins ,Cell Line ,law.invention ,03 medical and health sciences ,Prostate cancer ,0302 clinical medicine ,Ubiquitin ,Downregulation and upregulation ,law ,Cell Line, Tumor ,medicine ,Humans ,RNA-Seq ,Molecular Biology ,Actin ,TNF Receptor-Associated Factor 6 ,biology ,Chemistry ,Gene Expression Profiling ,Microfilament Proteins ,Ubiquitination ,Prostatic Neoplasms ,medicine.disease ,In vitro ,Ubiquitin ligase ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,PC-3 Cells ,Proteolysis ,biology.protein ,Cancer research ,Suppressor ,Signal transduction ,Protein Binding ,Signal Transduction - Abstract
Transgelin (TAGLN, also named SM22) is an actin-associated protein and affects dynamics of actin filaments. Deregulation of TAGLN contributes to the development of different cancers, and it is commonly considered to be a tumor suppressor. TAGLN is usually downregulated in prostate cancer; however, the detailed functions of TAGLN in prostate cancer and how TAGLN is regulated remains unclear. In this study, we confirmed that TAGLN is downregulated in prostate cancer tissues and demonstrated that the downregulation of TAGLN occurs through proteasomal degradation. Next, we found that the expression level of TAGLN is inversely correlated with TRAF6. We screened more than 20 E2–E3 pairs by in vitro ubiquitination assay and found that the E2A–TRAF6 pair catalyzed mono ubiquitination of TAGLN. We then identified the ubiquitination sites of TAGLN to be on K89 or K108 residues and demonstrated that ubiquitination of TAGLN on K89/K108 are important for TRAF6-mediated proteasomal degradation. Furthermore, we investigated the function of TAGLN in prostate cancer cells. We found that ablation of TAGLN promoted prostate cancer cell proliferation and suppressed their migration via activation of NF-κB and Myc signaling pathways. Overall, our study provided new insights into the mechanisms underlying TAGLN expression and activity in prostate cancer. Implications: E3 ligase TRAF6 mediate mono-ubiquitination and degradation of TAGLN, which leads to activation of NF-κB and Myc signaling pathways in prostate cancer cells. more...
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- 2021
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33. Prognostic value of glycolysis markers in head and neck squamous cell carcinoma: a meta-analysis
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Laibo Jiang, Bin Cheng, Xianyue Ren, Yuanyuan Li, Juan Xia, and Yanting Wang
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Monocarboxylic Acid Transporters ,Oncology ,Thyroid Hormones ,Aging ,medicine.medical_specialty ,Muscle Proteins ,PKM2 ,HNSCC ,glycolysis marker ,Hexokinase ,Internal medicine ,Biomarkers, Tumor ,medicine ,Humans ,Glycolysis ,Glucose Transporter Type 1 ,Glucose Transporter Type 4 ,biology ,business.industry ,Glucose transporter ,Membrane Proteins ,Cell Biology ,Prognosis ,medicine.disease ,Head and neck squamous-cell carcinoma ,meta-analysis ,stomatognathic diseases ,Head and Neck Neoplasms ,Carcinoma, Squamous Cell ,Monocarboxylate transporter 4 ,biology.protein ,GLUT1 ,Carrier Proteins ,business ,GLUT4 ,Pyruvate kinase ,Research Paper - Abstract
Glycolysis markers including glucose transporter 1 (GLUT1), monocarboxylate transporter 4 (MCT4), hexokinase 2 (HK2), pyruvate kinase M2 (PKM2) and glucose transporter 4 (GLUT4) play vital roles in head and neck squamous cell carcinoma (HNSCC). However, their prognostic value in HNSCC is still controversial. In this meta-analysis, we searched the PubMed, Web of Science and Cochrane Library databases and included thirty-seven studies (3272 patients) that met the inclusion criteria. Higher expression levels of the glycolysis markers in tumor tissues correlated with poorer overall survival (OS; P < 0.001), disease-free survival (DFS; P = 0.03) and recurrence-free survival (RFS; P < 0.001) of HNSCC patients. Subgroup and sensitivity analyses demonstrated that higher expression levels of GLUT1 (P < 0.001), MCT4 (P = 0.002), HK2 (P = 0.002) and PKM2 (P < 0.001) correlated with poorer OS among HNSCC patients. Higher expression of MCT4 (P < 0.001) and PKM2 (P = 0.008) predicted poorer DFS among HNSCC patients. However, GLUT4 expression levels did not associate with clinical outcomes in HNSCC patients. These results demonstrate that glycolysis markers, such as GLUT1, MCT4, HK2 and PKM2, are potential prognostic predictors and therapeutic targets in HNSCC. more...
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- 2021
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34. Classification of node-positive melanomas into prognostic subgroups using keratin, immune, and melanogenesis expression patterns
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Eran Nizri, Dvir Netanely, Dov Hershkovitz, Stav Leibou, Tomer Perluk, Sarah Amar, Hananya Vaknine, Ronen Brenner, Ron Shamir, Jacob Frand, Valentina Zemser-Werner, Rivi Haiat Factor, Carmit Levy, Roma Parikh, and Neta Stern more...
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Male ,0301 basic medicine ,Cancer Research ,Carcinogenesis ,Ubiquitin-Protein Ligases ,Muscle Proteins ,Disease ,Biology ,Article ,Metastasis ,Tripartite Motif Proteins ,Tumour biomarkers ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Keratin ,Cancer genomics ,Genetics ,medicine ,Humans ,RNA-Seq ,Neoplasm Metastasis ,Receptors, Immunologic ,Melanoma ,Molecular Biology ,Gene ,Melanosome ,Melanins ,chemistry.chemical_classification ,Melanosomes ,Immunity ,medicine.disease ,Survival Analysis ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,Cutaneous melanoma ,Cancer research ,Female ,Kallikreins ,Transcriptome - Abstract
Cutaneous melanoma tumors are heterogeneous and show diverse responses to treatment. Identification of robust molecular biomarkers for classifying melanoma tumors into clinically distinct and homogenous subtypes is crucial for improving the diagnosis and treatment of the disease. In this study, we present a classification of melanoma tumors into four subtypes with different survival profiles based on three distinct gene expression signatures: keratin, immune, and melanogenesis. The melanogenesis expression pattern includes several genes that are characteristic of the melanosome organelle and correlates with worse survival, suggesting the involvement of melanosomes in melanoma aggression. We experimentally validated the secretion of melanosomes into surrounding tissues by melanoma tumors, which potentially affects the lethality of metastasis. We propose a simple molecular decision tree classifier for predicting a tumor’s subtype based on representative genes from the three identified signatures. Key predictor genes were experimentally validated on melanoma samples taken from patients with varying survival outcomes. Our three-pattern approach for classifying melanoma tumors can contribute to advancing the understanding of melanoma variability and promote accurate diagnosis, prognostication, and treatment. more...
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- 2021
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35. Quantitative single-protein imaging reveals molecular complex formation of integrin, talin, and kindlin during cell adhesion
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Lisa S. Fischer, Ralph T. Böttcher, Thomas Schlichthaerle, Reinhard Fässler, C. Klingner, Maximilian T. Strauss, Ralf Jungmann, and Carsten Grashoff
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0301 basic medicine ,Talin ,Integrins ,In silico ,Science ,Complex formation ,Integrin ,General Physics and Astronomy ,Muscle Proteins ,02 engineering and technology ,General Biochemistry, Genetics and Molecular Biology ,Article ,Cell Line ,03 medical and health sciences ,Mice ,Microscopy ,Cell Adhesion ,Animals ,Humans ,Super-resolution microscopy ,Cell adhesion ,Multidisciplinary ,biology ,Chemistry ,Integrin beta1 ,Resolution (electron density) ,General Chemistry ,Adhesion ,021001 nanoscience & nanotechnology ,Single Molecule Imaging ,Focal adhesion ,Cytoskeletal Proteins ,030104 developmental biology ,Biophysics ,biology.protein ,0210 nano-technology ,Molecular probe - Abstract
Single-molecule localization microscopy (SMLM) enabling the investigation of individual proteins on molecular scales has revolutionized how biological processes are analysed in cells. However, a major limitation of imaging techniques reaching single-protein resolution is the incomplete and often unknown labeling and detection efficiency of the utilized molecular probes. As a result, fundamental processes such as complex formation of distinct molecular species cannot be reliably quantified. Here, we establish a super-resolution microscopy framework, called quantitative single-molecule colocalization analysis (qSMCL), which permits the identification of absolute molecular quantities and thus the investigation of molecular-scale processes inside cells. The method combines multiplexed single-protein resolution imaging, automated cluster detection, in silico data simulation procedures, and widely applicable experimental controls to determine absolute fractions and spatial coordinates of interacting species on a true molecular level, even in highly crowded subcellular structures. The first application of this framework allowed the identification of a long-sought ternary adhesion complex—consisting of talin, kindlin and active β1-integrin—that specifically forms in cell-matrix adhesion sites. Together, the experiments demonstrate that qSMCL allows an absolute quantification of multiplexed SMLM data and thus should be useful for investigating molecular mechanisms underlying numerous processes in cells., Single-molecule localisation microscopy is limited by low labeling and detection efficiencies of the molecular probes. Here the authors report a framework to obtain absolute molecular quantities on a true molecular scale; the data reveal a ternary adhesion complex underlying cell-matrix adhesion. more...
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- 2021
36. Eicosapentaenoic acid changes muscle transcriptome and intervenes in aging-related fiber type transition in male mice
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Noritada Yoshikawa, Masaaki Uehara, Mayu Nishimura, Ken Ichirou Morohashi, Akiko Kuribara-Souta, Motohisa Yamamoto, Hiroki Yamazaki, and Hirotoshi Tanaka
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Male ,Aging ,medicine.medical_specialty ,Physiology ,Endocrinology, Diabetes and Metabolism ,Muscle Proteins ,Biology ,complex mixtures ,Transcriptome ,Mice ,Physiology (medical) ,Internal medicine ,medicine ,Animals ,health care economics and organizations ,Fiber type ,Transition (genetics) ,Skeletal muscle ,Muscle weakness ,social sciences ,medicine.disease ,Eicosapentaenoic acid ,Phenotype ,Mice, Inbred C57BL ,Muscle Fibers, Slow-Twitch ,Endocrinology ,medicine.anatomical_structure ,Eicosapentaenoic Acid ,Gene Expression Regulation ,Sarcopenia ,Muscle Fibers, Fast-Twitch ,lipids (amino acids, peptides, and proteins) ,Insulin Resistance ,medicine.symptom ,geographic locations ,Research Article - Abstract
Age-related sarcopenia is associated with a variety of changes in skeletal muscle. These changes are interrelated with each other and associated with systemic metabolism, the details of which, however, are largely unknown. Eicosapentaenoic acid (EPA) is a promising nutrient against sarcopenia and has multifaceted effects on systemic metabolism. In this study, we hypothesized that the aging process in skeletal muscle can be intervened by the administration of EPA. Seventy-five-week-old male mice were assigned to groups fed an EPA-deprived diet (EPA−) or an EPA-enriched diet with 1 wt% EPA (EPA+) for 12 wk. Twenty-four-week-old male mice fed with normal chow were also analyzed. At baseline, the grip strength of the aging mice was lower than that of the young mice. After 12 wk, EPA+ showed similar muscle mass but increased grip strength compared with EPA−. EPA+ displayed higher insulin sensitivity than EPA−. Immunohistochemistry and gene expression analysis of myosin heavy chains (MyHCs) revealed fast-to-slow fiber type transition in aging muscle, which was partially inhibited by EPA. RNA sequencing (RNA-Seq) analysis suggested that EPA supplementation exerts pathway-specific effects in skeletal muscle including the signatures of slow-to-fast fiber type transition. In conclusion, we revealed that aging skeletal muscle in male mice shows lower grip strength and fiber type changes, both of which can be inhibited by EPA supplementation irrespective of muscle mass alteration. NEW & NOTEWORTHY This study demonstrated that the early phenotype of skeletal muscle in aging male mice is characterized by muscle weakness with fast-to-slow fiber type transition, which could be ameliorated by feeding with EPA-enriched diet. EPA induced metabolic changes such as an increase in systemic insulin sensitivity and altered muscle transcriptome in the aging mice. These changes may be related to the fiber type transition and influence muscle quality. more...
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- 2021
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37. CircTM7SF3 contributes to oxidized low-density lipoprotein-induced apoptosis, inflammation and oxidative stress through targeting miR-206/ASPH axis in atherosclerosis cell model in vitro
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Ming Bai and Xiaojuan Wang
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Male ,lcsh:Diseases of the circulatory (Cardiovascular) system ,ASPH ,THP-1 Cells ,Cell ,Muscle Proteins ,Apoptosis ,medicine.disease_cause ,Mixed Function Oxygenases ,0302 clinical medicine ,0303 health sciences ,biology ,medicine.diagnostic_test ,miR-206 ,Middle Aged ,Lipoproteins, LDL ,medicine.anatomical_structure ,circTM7SF3 ,030220 oncology & carcinogenesis ,Female ,medicine.symptom ,Inflammation Mediators ,Cardiology and Cardiovascular Medicine ,Signal Transduction ,Research Article ,Inflammation ,Flow cytometry ,Ox-LDL ,03 medical and health sciences ,medicine ,Humans ,Viability assay ,030304 developmental biology ,Aged ,Messenger RNA ,business.industry ,Interleukin-6 ,Tumor Necrosis Factor-alpha ,Macrophages ,Calcium-Binding Proteins ,Membrane Proteins ,RNA, Circular ,Atherosclerosis ,Molecular biology ,MicroRNAs ,Oxidative Stress ,Gene Expression Regulation ,lcsh:RC666-701 ,Case-Control Studies ,biology.protein ,business ,Oxidative stress - Abstract
Background Atherosclerosis (AS) is a chronic inflammatory disorder. The aim of our study was to explore the role of circular RNA (circRNA) transmembrane 7 superfamily member 3 (circTM7SF3) in AS progression. Methods Experiments were conducted using oxidized low-density lipoprotein (ox-LDL)-induced THP-1-derived macrophages and differentiated human monocyte-derived macrophages (hMDMs). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of circTM7SF3, its linear form TM7SF3, microRNA-206 (miR-206) and aspartyl (asparaginyl) β-hydroxylase (ASPH) messenger RNA (mRNA). Cell viability and apoptosis were examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. Cell inflammation was analyzed by measuring the production of tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) using enzyme-linked immunosorbent assay (ELISA) kits. Cell oxidative stress was assessed through analyzing the levels of oxidative stress markers using their corresponding commercial kits. Dual-luciferase reporter assay and RNA-pull down assay were used to confirm the interaction between miR-206 and circTM7SF3 or ASPH. The protein level of ASPH was examined by Western blot assay. Results CircTM7SF3 level was markedly increased in the serum samples of AS patients and ox-LDL-induced THP-1-derived macrophages compared with their matching counterparts. ox-LDL induced-damage in THP-1 cells was partly attenuated by the interference of circTM7SF3. MiR-206 was a downstream molecular target of circTM7SF3. Si-circTM7SF3-mediated effects in ox-LDL-induced THP-1-derived macrophages were partly ameliorated by the addition of anti-miR-206. MiR-206 directly interacted with ASPH mRNA. CircTM7SF3 silencing reduced the expression of ASPH partly through up-regulating miR-206 in THP-1-derived macrophages. ASPH overexpression partly counteracted the effects induced by miR-206 overexpression in ox-LDL-induced THP-1-derived macrophages. Conclusion CircTM7SF3 contributed to ox-LDL-induced injury in AS cell model through up-regulating the expression of ASPH via targeting miR-206. more...
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- 2021
38. A Dominant C150Y Mutation in FHL1 Induces Structural Alterations in LIM2 Domain Causing Protein Aggregation In Human and Drosophila Indirect Flight Muscles
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Veeramani Preethish-Kumar, Upendra Nongthomba, Keshava Prasad T S, Kiran K. Mangalaparthi, Sruthi Unni, Nalini Atchayaram, Balasundaram Padmanabhan, Rashmi Santhoshkumar, and Gayathri Narayanappa
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Adult ,Male ,Protein Conformation, alpha-Helical ,0301 basic medicine ,Mutant ,Mutation, Missense ,Muscle Proteins ,Protein aggregation ,medicine.disease_cause ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,Muscular Diseases ,Protein Domains ,medicine ,Animals ,Humans ,Child ,Muscle, Skeletal ,Exome sequencing ,Genes, Dominant ,Mutation ,Muscle biopsy ,biology ,medicine.diagnostic_test ,Intracellular Signaling Peptides and Proteins ,Genetic Diseases, X-Linked ,General Medicine ,LIM Domain Proteins ,biology.organism_classification ,Phenotype ,FHL1 ,Cell biology ,Drosophila melanogaster ,030104 developmental biology ,Female ,Protein Multimerization ,030217 neurology & neurosurgery - Abstract
FHL1-related myopathies are rare X-linked dominant myopathies. Though clinically classified into several subgroups, spinal and scapuloperoneal muscle involvement are common to all. In this study, we identified c.449G > A, p.C150Y mutation by clinical exome sequencing in two patients from same family (son and mother) of Indian origin who presented with multiple contractures. Muscle biopsy showed numerous intracytoplasmic aggregates intensely stained on HE and MGT. The strong reactions to M-NBT revealed aggregates to be reducing bodies and positively labeled to anti-FHL1 antibody. Ultrastructurally, Z-band streaming and granular and granulofilamentous material were seen. Further, the translational evidence of mutant peptide was confirmed using mass spectrometric analysis. To establish p.C150Y as the cause for protein aggregation, in vivo studies were carried out using transgenic Drosophila model which highlighted Z-band abnormalities and protein aggregates in indirect flight muscles with compromised physiological function. Thus, recapitulating the X-linked human disease phenotype. Additionally, the molecular dynamics simulation analysis unraveled the drastic change in α-helix of LIM2, the region immediately next to site of C150Y mutation that could be the plausible cause for protein aggregation. To the best of our knowledge, this is the first study of p.C150Y mutation in FHL1 identified in Indian patients with in vivo and in silico analysis to establish the cause for protein aggregation in muscle. more...
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- 2021
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39. Systemic effect of FHL1 on neuromuscular junction and myotube formation via insulin-like growth factor and myostatin signaling pathways
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Kai Zhao, Wei Jiang, Feixu Zhang, Xia Wu, Jiamei Wu, Zengmin Du, Ying Chen, Jing Zheng, Chengyong Shen, and Xiao Xiao
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0301 basic medicine ,Scaffold protein ,Follistatin ,Muscle Fibers, Skeletal ,Neuromuscular Junction ,Biophysics ,Muscle Proteins ,Myostatin ,Biochemistry ,Neuromuscular junction ,Cell Line ,Cell Fusion ,Mice ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Receptors, Cholinergic ,Phosphorylation ,Molecular Biology ,Cell Proliferation ,Acetylcholine receptor ,biology ,Myogenesis ,Intracellular Signaling Peptides and Proteins ,Receptor Protein-Tyrosine Kinases ,Cell Differentiation ,Cell Biology ,LIM Domain Proteins ,FHL1 ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,biology.protein ,Signal transduction ,Signal Transduction - Abstract
Four-and-a-half LIM domain protein 1 (FHL1) is a member of the FHL protein family that serves as a scaffold protein to maintain normal cellular structure and function. Its mutations have been implicated in multiple muscular diseases. These FHL1 related myopathies are characterized by symptoms such as progressive muscle loss, rigid or bent spine, even cardiac or respiratory failure in some patients, which implies pathological problems not only in muscles, but also in the nervous system. Moreover, decreased FHL1 protein level has been found in patients with FHL1 mutations, indicating the protein loss-of-function as a pathological cause of such diseases. These findings suggest the significance of understanding the systemic role of FHL1 in the homeostasis of nervous system and muscle. Here we reported that Fhl1 loss in C2C12 myotubes obscured acetylcholine receptor (AChR) clustering in addition to myotube fusion, which was associated with impaired MuSK phosphorylation. Mechanistically, myostatin-SMAD2/3 signaling was enhanced, whereas IGF-PI3K-AKT signaling was suppressed in Fhl1-/- C2C12 myotubes. Reversion of these molecular alterations rescued AChR clustering and differentiation deficits. These data outline a systemic regulation of AChR clustering and myotube fusion by FHL1, which may offer clues for mechanism study and development of therapeutic strategies to treat FHL1 related myopathies. more...
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- 2021
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40. Mutational Spectrum of CAPN3 with Genotype-Phenotype Correlations in Limb Girdle Muscular Dystrophy Type 2A/R1 (LGMD2A/LGMDR1) Patients in India
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Pankaj Jha, Mohammed Faruq, Mohammad Husain, Mehar Chand Sharma, Pankaj Pathak, Sumit Randhir Singh, Chitra Sarkar, Vaishali Suri, Prerana Jha, Rohit Bhatia, and Sheffali Gulati
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Population ,India ,Muscle Proteins ,Biology ,Carrier testing ,medicine ,Humans ,Missense mutation ,Genetic Testing ,Muscular dystrophy ,Allele ,education ,Myopathy ,Genetic Association Studies ,Genetic testing ,Genetics ,education.field_of_study ,medicine.diagnostic_test ,Calpain ,Sequence Analysis, DNA ,medicine.disease ,Muscular Dystrophies, Limb-Girdle ,Neurology ,Mutation ,Neurology (clinical) ,medicine.symptom ,Limb-girdle muscular dystrophy - Abstract
Background: Limb girdle muscular dystrophy recessive type 1 (LGMDR1, Previously LGMD2A) is characterized by inactivating mutations in CAPN3. Despite the significant burden of muscular dystrophy in India, and particularly of LGMDR1, its genetic characterization and possible phenotypic manifestations are yet unidentified. Material and Methods: We performed bidirectional CAPN3 sequencing in 95 LGMDR1 patient samples characterized by calpain-3 protein analysis, and these findings were correlated with clinical, biochemical and histopathological features. Results: We identified 84 (88.4%) cases of LGMDR1 harboring 103 CAPN3 mutations (71 novel and 32 known). At least two mutant alleles were identified in 79 (94.2%) of patients. Notably, 76% exonic variations were enriched in nine CAPN3 exons and overall, 41 variations (40%) correspond to only eight exonic and intronic mutations. Patients with two nonsense/out of frame/splice-site mutations showed significant loss of calpain-3 protein as compared to those with two missense/inframe mutations (P = 0.04). We observed a slow progression of disease and less severity in our patients compared to European population. Rarely, presenting clinical features were atypical, and mimicked other muscle diseases like FSHMD, distal myopathy and metabolic myopathies. Conclusion: This is first systematic study to characterize the genetic framework of LGMDR1 in the Indian population. Preliminary calpain-3 immunoblot screening serves well to direct genetic testing. Our findings prioritized nine CAPN3 exons for LGMDR1 diagnosis in our population; therefore, a targeted-sequencing panel of nine exons could serve well for genetic diagnosis, carrier testing, counseling and clinical trial feasibility study in LGMDR1 patients in India. more...
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- 2021
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41. A functional family of fluorescent nucleotide analogues to investigate actin dynamics and energetics
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Jenna Elliott, Jessica Colombo, Pekka Lappalainen, Konstantin Kogan, Alphée Michelot, Tommi Kotila, Audrey Guillotin, Adrien Antkowiak, Institut de Biologie du Développement de Marseille (IBDM), Aix Marseille Université (AMU)-Collège de France (CdF (institution))-Centre National de la Recherche Scientifique (CNRS), University of Helsinki, Helsingin yliopisto = Helsingfors universitet = University of Helsinki, and Institute of Biotechnology more...
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0301 basic medicine ,Muscle Proteins ,General Physics and Astronomy ,Plasma protein binding ,[SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC] ,0302 clinical medicine ,BINDING ,Nucleotide ,ComputingMilieux_MISCELLANEOUS ,chemistry.chemical_classification ,Multidisciplinary ,biology ,[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,Nucleotides ,Chemistry ,Hydrolysis ,Microfilament Proteins ,Energetics ,Fluorescence ,Actin Cytoskeleton ,POLYMERIZATION ,Profilin ,Thermodynamics ,Rabbits ,Algorithms ,Protein Binding ,NUCLEATION ,Science ,Kinetics ,Fluorescence Polarization ,macromolecular substances ,Article ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Animals ,Actin ,Fluorescent Dyes ,COMPLEX ,General Chemistry ,Models, Theoretical ,Actins ,FILAMENT TURNOVER ,030104 developmental biology ,PROFILIN ,Cellular motility ,biology.protein ,Biophysics ,VISUALIZATION ,1182 Biochemistry, cell and molecular biology ,030217 neurology & neurosurgery ,Fluorescence anisotropy - Abstract
Actin polymerization provides force for vital processes of the eukaryotic cell, but our understanding of actin dynamics and energetics remains limited due to the lack of high-quality probes. Most current probes affect dynamics of actin or its interactions with actin-binding proteins (ABPs), and cannot track the bound nucleotide. Here, we identify a family of highly sensitive fluorescent nucleotide analogues structurally compatible with actin. We demonstrate that these fluorescent nucleotides bind to actin, maintain functional interactions with a number of essential ABPs, are hydrolyzed within actin filaments, and provide energy to power actin-based processes. These probes also enable monitoring actin assembly and nucleotide exchange with single-molecule microscopy and fluorescence anisotropy kinetics, therefore providing robust and highly versatile tools to study actin dynamics and functions of ABPs., Actin polymerization provides force for vital processes of the eukaryotic cell, but our understanding of actin dynamics and energetics remains limited due to the lack of high-quality probes. Here authors identify a family of highly sensitive fluorescent nucleotide analogues which bind to actin and provide energy to power actin-based processes. more...
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- 2021
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42. Initiation of focal adhesion assembly by talin and kindlin: A dynamic view
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Edward F. Plow, Liang Zhu, and Jun Qin
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Models, Molecular ,Talin ,Integrins ,Integrin ,Muscle Proteins ,Focal adhesion assembly ,Review ,Biochemistry ,Focal adhesion ,Mice ,03 medical and health sciences ,Interaction network ,Cell surface receptor ,Animals ,Cell adhesion ,Molecular Biology ,030304 developmental biology ,Mice, Knockout ,Focal Adhesions ,0303 health sciences ,biology ,Chemistry ,030302 biochemistry & molecular biology ,Cell migration ,Cell biology ,Cytoskeletal Proteins ,biology.protein ,Function (biology) ,Protein Binding ,Signal Transduction - Abstract
Focal adhesions (FAs) are integrin-containing protein complexes regulated by a network of hundreds of protein-protein interactions. They are formed in a spatiotemporal manner upon the activation of integrin transmembrane receptors, which is crucial to trigger cell adhesion and many other cellular processes including cell migration, spreading and proliferation. Despite decades of studies, a detailed molecular level understanding on how FAs are organized and function is lacking due to their highly complex and dynamic nature. However, advances have been made on studying key integrin activators, talin and kindlin, and their associated proteins, which are major components of nascent FAs critical for initiating the assembly of mature FAs. This review will discuss the structural and functional findings of talin and kindlin and their immediate interaction network, which will shed light upon the architecture of nascent FAs and how they act as seeds for FA assembly to dynamically regulate diverse adhesion-dependent physiological and pathological responses. more...
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- 2020
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43. SPEG binds with desmin and its deficiency causes defects in triad and focal adhesion proteins
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Isabelle Marty, Jasmine Lin, Pankaj B. Agrawal, Yuanfan Zhang, Qifei Li, Shiyu Luo, Shideh Kazerounian, and Quinn Murphy
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Male ,Myosin light-chain kinase ,Integrin ,Muscle Proteins ,Mice, Transgenic ,Biology ,Desmin ,Focal adhesion ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Genetics ,Animals ,Muscle, Skeletal ,Myosin-Light-Chain Kinase ,Molecular Biology ,Genetics (clinical) ,030304 developmental biology ,Mice, Knockout ,RYR1 ,Focal Adhesions ,0303 health sciences ,Intracellular Signaling Peptides and Proteins ,General Medicine ,Vinculin ,Cell biology ,Triadin ,Mutation ,biology.protein ,Calcium ,General Article ,Myofibril ,Cell Adhesion Molecules ,030217 neurology & neurosurgery ,Myopathies, Structural, Congenital - Abstract
Striated preferentially expressed gene (SPEG), a member of the myosin light chain kinase family, is localized at the level of triad surrounding myofibrils in skeletal muscles. In humans, SPEG mutations are associated with centronuclear myopathy and cardiomyopathy. Using a striated muscle-specific Speg-knockout (KO) mouse model, we have previously shown that SPEG is critical for triad maintenance and calcium handling. Here, we further examined the molecular function of SPEG and characterized the effects of SPEG deficiency on triad and focal adhesion proteins. We used yeast two-hybrid assay, and identified desmin, an intermediate filament protein, to interact with SPEG and confirmed this interaction by co-immunoprecipitation. Using domain-mapping assay, we defined that Ig-like and fibronectin III domains of SPEG interact with rod domain of desmin. In skeletal muscles, SPEG depletion leads to desmin aggregates in vivo and a shift in desmin equilibrium from soluble to insoluble fraction. We also profiled the expression and localization of triadic proteins in Speg-KO mice using western blot and immunofluorescence. The amount of RyR1 and triadin were markedly reduced, whereas DHPRα1, SERCA1 and triadin were abnormally accumulated in discrete areas of Speg-KO myofibers. In addition, Speg-KO muscles exhibited internalized vinculin and β1 integrin, both of which are critical components of the focal adhesion complex. Further, β1 integrin was abnormally accumulated in early endosomes of Speg-KO myofibers. These results demonstrate that SPEG-deficient skeletal muscles exhibit several pathological features similar to those seen in MTM1 deficiency. Defects of shared cellular pathways may underlie these structural and functional abnormalities in both types of diseases. more...
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- 2020
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44. NSUN5 Facilitates Viral RNA Recognition by RIG-I Receptor
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Jiaqian Liang, Chen Wang, Wei Meng, Chenhui Li, Haoyang Zeng, Haiyang Hu, Quanyi Wang, Juanjuan Zhu, Boyue Sun, Lele Zhang, and Fuqiang Ye
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viruses ,Immunology ,Muscle Proteins ,chemical and pharmacologic phenomena ,Mice ,RNA Virus Infections ,Chlorocebus aethiops ,Animals ,Humans ,RNA Viruses ,Immunology and Allergy ,Viral rna ,Receptors, Immunologic ,Receptor ,Vero Cells ,Gene ,RNA, Double-Stranded ,tRNA Methyltransferases ,biology ,RIG-I ,virus diseases ,RNA ,RNA virus ,Methyltransferases ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,Immunity, Innate ,Cell biology ,RNA silencing ,HEK293 Cells ,DEAD Box Protein 58 ,RNA, Viral ,biological phenomena, cell phenomena, and immunity ,Signal transduction - Abstract
The RIG-I receptor induces the innate antiviral responses upon sensing RNA viruses. The mechanisms through which RIG-I optimizes the strength of the downstream signaling remain incompletely understood. In this study, we identified that NSUN5 could potentiate the RIG-I innate signaling pathway. Deficiency of NSUN5 enhanced RNA virus proliferation and inhibited the induction of the downstream antiviral genes. Consistently, NSUN5-deficient mice were more susceptible to RNA virus infection than their wild-type littermates. Mechanistically, NSUN5 bound directly to both viral RNA and RIG-I, synergizing the recognition of dsRNA by RIG-I. Collectively, to our knowledge, this study characterized NSUN5 as a novel RIG-I coreceptor, playing a vital role in restricting RNA virus infection. more...
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- 2020
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45. Alternative splicing transitions associate with emerging atrophy phenotype during denervation‐induced skeletal muscle atrophy
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Jiaying Qiu, Junjie Sun, Hualin Sun, Yan Chang, and Liucheng Wu
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Male ,0301 basic medicine ,Time Factors ,Physiology ,Clinical Biochemistry ,Muscle Proteins ,Biology ,Cell Line ,Rats, Sprague-Dawley ,Mice ,03 medical and health sciences ,Exon ,0302 clinical medicine ,Atrophy ,Tibialis anterior muscle ,medicine ,Animals ,Guanine Nucleotide Exchange Factors ,RNA-Seq ,Muscle, Skeletal ,Cell Proliferation ,Denervation ,Gene Expression Profiling ,Alternative splicing ,Cell Differentiation ,Cell Biology ,medicine.disease ,Sciatic Nerve ,Phenotype ,Muscle Denervation ,Muscle atrophy ,Cell biology ,Alternative Splicing ,Disease Models, Animal ,Muscular Atrophy ,030104 developmental biology ,030220 oncology & carcinogenesis ,medicine.symptom ,Transcriptome ,C2C12 - Abstract
Alternative splicing (AS) presents a key posttranscriptional regulatory mechanism associated with numerous physiological processes. However, little is known about its role in skeletal muscle atrophy. In this study, we used a rat model of denervated skeletal muscle atrophy and performed RNA-sequencing to analyze transcriptome profiling of tibialis anterior muscle at multiple time points following denervation. We found that AS is a novel mechanism involving muscle atrophy, which is independent changes at the transcript level. Bioinformatics analysis further revealed that AS transitions are associated with the appearance of the atrophic phenotype. Moreover, we found that the inclusion of multiple highly conserved exons of Obscn markedly increased at 3 days after denervation. In addition, we confirmed that this newly transcript inhibited C2C12 cell proliferation and exacerbated myotube atrophy. Finally, our study revealed that a large number of RNA-binding proteins were upregulated when the atrophy phenotype appeared. Our data emphasize the importance of AS in this process. more...
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- 2020
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46. Soluble RANKL exaggerates hindlimb suspension‐induced osteopenia but not muscle protein balance
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Toni L. Speacht, Charles H. Lang, and Henry J. Donahue
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Male ,musculoskeletal diseases ,medicine.medical_specialty ,0206 medical engineering ,Protein metabolism ,Muscle Proteins ,P70-S6 Kinase 1 ,02 engineering and technology ,Article ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Internal medicine ,Cortical Bone ,medicine ,Animals ,Orthopedics and Sports Medicine ,Femur ,Receptor ,030203 arthritis & rheumatology ,biology ,RANK Ligand ,medicine.disease ,020601 biomedical engineering ,Mice, Inbred C57BL ,Osteopenia ,Bone Diseases, Metabolic ,Endocrinology ,medicine.anatomical_structure ,Hindlimb Suspension ,chemistry ,RANKL ,Sarcopenia ,biology.protein ,Cortical bone - Abstract
We examined the hypothesis that exaggerating unloading-induced bone loss using a combination of hindlimb suspension (HLS) and exogenous injections of receptor activator of nuclear factor kappa-B ligand (RANKL) also exaggeratesgastrocnemius and quadriceps muscle loss. Forty, male C57Bl/6J mice (16 weeks) were subjected to HLS or normal ambulation (ground control, GC) for 14 days. Mice received 3 intraperitoneal injections of either human recombinant soluble RANKL or PBS as control (n=10/group) at 24 hour intervals starting on Day 1 of HLS. GC + RANKL and HLS mice exhibited similar decreases in trabecular bone volume and density in both proximal tibias and distal femurs. However, RANKL affected trabecular number, separation, and connectivity density, while HLS decreased trabecular thickness. The combination of RANKL and HLS exacerbated these changes. Similarly,GC + RANKL and HLS mice saw comparable decreases in cortical bone volume, thickness, and strength in femur midshafts, and combination treatment exacerbated these changes. Plasma concentrations of P1NP were increased in both groups receiving RANKL, while CTX concentrations were unchanged.HLS decreased gastrocnemius weight and was associated with a reduction in global protein synthesis, and no change in proteasome activity. This change was correlated with a decrease in S6K1 and S6 phosphorylation, but no change in 4E-BP1 phosphorylation. Injection of RANKL did not alter gastrocnemius or quadriceps muscle protein metabolism in GC or HLS mice.Our results suggest that injection of soluble RANKL exacerbates unloading-induced bone loss, but not unloading-induced gastrocnemius or quadriceps muscle loss. This article is protected by copyright. All rights reserved. more...
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- 2020
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47. Nuclear numbers in syncytial muscle fibers promote size but limit the development of larger myonuclear domains
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Douglas P. Millay, Einar Eftestøl, Alyssa A. W. Cramer, Julien Ochala, Sakthivel Sadayappan, Vikram Prasad, Hannah F. Dugdale, Taejeong Song, Kristian Gundersen, and Kenth-Arne Hansson
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Male ,0301 basic medicine ,Satellite Cells, Skeletal Muscle ,Science ,Skeletal muscle ,Muscle Proteins ,General Physics and Astronomy ,Mice, Transgenic ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Developmental biology ,medicine ,Animals ,Muscle, Skeletal ,Cell Size ,Cell Nucleus ,Multidisciplinary ,Extramural ,Membrane Proteins ,General Chemistry ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Models, Animal ,Reserve capacity ,Female ,030217 neurology & neurosurgery - Abstract
Mammalian cells exhibit remarkable diversity in cell size, but the factors that regulate establishment and maintenance of these sizes remain poorly understood. This is especially true for skeletal muscle, comprised of syncytial myofibers that each accrue hundreds of nuclei during development. Here, we directly explore the assumed causal relationship between multinucleation and establishment of normal size through titration of myonuclear numbers during mouse neonatal development. Three independent mouse models, where myonuclear numbers were reduced by 75, 55, or 25%, led to the discovery that myonuclei possess a reserve capacity to support larger functional cytoplasmic volumes in developing myofibers. Surprisingly, the results revealed an inverse relationship between nuclei numbers and reserve capacity. We propose that as myonuclear numbers increase, the range of transcriptional return on a per nuclear basis in myofibers diminishes, which accounts for both the absolute reliance developing myofibers have on nuclear accrual to establish size, and the limits of adaptability in adult skeletal muscle., Skeletal muscle is composed of syncytial myofibres, each containing hundreds of nuclei. Through genetic reduction of the number of nuclei per myofibre, the authors confirm that more nuclei produce larger cells but myofibres with fewer nuclei adaptively compensate leading to larger and functional myonuclear domains. more...
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- 2020
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48. Mitochondrial dysfunction and consequences in calpain-3-deficient muscle
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Jack H. Van der Meulen, Jennifer M. Peterson, Helen Johnston, Aditi Phadke, Jessica F. Boehler, Aurelia Defour, Vanessa E. Jahnke, Kanneboyina Nagaraju, Qing Yu, Jyoti K. Jaiswal, and Kitipong Uaesoontrachoon more...
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0301 basic medicine ,lcsh:Diseases of the musculoskeletal system ,Muscle Proteins ,Mitochondrion ,Sarcomere ,Cell Line ,Myoblasts ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Loss of Function Mutation ,medicine ,Animals ,Myocyte ,Orthopedics and Sports Medicine ,PPAR delta ,Muscular dystrophy ,Molecular Biology ,Cells, Cultured ,MyoD Protein ,Organelle Biogenesis ,biology ,Calpain ,Muscle cell differentiation ,Research ,PAX7 Transcription Factor ,Skeletal muscle ,Cell Biology ,medicine.disease ,Mitochondria, Muscle ,Cell biology ,Mitochondria ,Mice, Inbred C57BL ,Thiazoles ,030104 developmental biology ,medicine.anatomical_structure ,Mitochondrial biogenesis ,biology.protein ,Calpain-3 deficiency ,Muscle membrane repair ,lcsh:RC925-935 ,030217 neurology & neurosurgery ,LGMD2A - Abstract
BackgroundNonsense or loss-of-function mutations in the non-lysosomal cysteine protease calpain-3 result in limb-girdle muscular dystrophy type 2A (LGMD2A). While calpain-3 is implicated in muscle cell differentiation, sarcomere formation, and muscle cytoskeletal remodeling, the physiological basis for LGMD2A has remained elusive.MethodsCell growth, gene expression profiling, and mitochondrial content and function were analyzed using muscle and muscle cell cultures established from healthy and calpain-3-deficient mice. Calpain-3-deficient mice were also treated with PPAR-delta agonist (GW501516) to assess mitochondrial function and membrane repair. The unpairedttest was used to assess the significance of the differences observed between the two groups or treatments. ANOVAs were used to assess significance over time.ResultsWe find that calpain-3 deficiency causes mitochondrial dysfunction in the muscles and myoblasts. Calpain-3-deficient myoblasts showed increased proliferation, and their gene expression profile showed aberrant mitochondrial biogenesis. Myotube gene expression analysis further revealed altered lipid metabolism in calpain-3-deficient muscle. Mitochondrial defects were validated in vitro and in vivo. We used GW501516 to improve mitochondrial biogenesis in vivo in 7-month-old calpain-3-deficient mice. This treatment improved satellite cell activity as indicated by increased MyoD and Pax7 mRNA expression. It also decreased muscle fatigability and reduced serum creatine kinase levels. The decreased mitochondrial function also impaired sarcolemmal repair in the calpain-3-deficient skeletal muscle. Improving mitochondrial activity by acute pyruvate treatment improved sarcolemmal repair.ConclusionOur results provide evidence that calpain-3 deficiency in the skeletal muscle is associated with poor mitochondrial biogenesis and function resulting in poor sarcolemmal repair. Addressing this deficit by drugs that improve mitochondrial activity offers new therapeutic avenues for LGMD2A. more...
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- 2020
49. Hypermethylation-mediated silencing of NDRG4 promotes pancreatic ductal adenocarcinoma by regulating mitochondrial function
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Xing-Jing Luo, Hai-E Liu, and Hao-Hong Shi
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China ,Epithelial-Mesenchymal Transition ,endocrine system diseases ,Gene Expression ,Muscle Proteins ,Apoptosis ,Nerve Tissue Proteins ,Mitochondrion ,Biology ,Adenocarcinoma ,Biochemistry ,Methylation ,Article ,Epigenesis, Genetic ,03 medical and health sciences ,Cell Movement ,Cell Line, Tumor ,Databases, Genetic ,Biomarkers, Tumor ,Gene silencing ,Humans ,NDRG4 ,Neoplasm Invasiveness ,Epigenetics ,Molecular Biology ,Cell Proliferation ,Regulation of gene expression ,0303 health sciences ,Gene knockdown ,Cell growth ,030302 biochemistry & molecular biology ,Cell Cycle ,PDAC ,General Medicine ,DNA Methylation ,digestive system diseases ,Mitochondria ,Gene Expression Regulation, Neoplastic ,Cancer cell ,DNA methylation ,Cancer research ,Mitochondrial function ,Carcinoma, Pancreatic Ductal - Abstract
The N-myc downstream regulated gene (NDRG) family members are dysregulated in several tumors. Functionally, NDRGs play an important role in the malignant progression of cancer cells. However, little is known about the potential implications of NDRG4 in pancreatic ductal adenocarcinoma (PDAC). The aim of the current study was to elucidate the expression pattern of NDRG4 in PDAC and evaluate its potential cellular biological effects. Here, we firstly report that epigenetic-mediated silencing of NDRG4 promotes PDAC by regulating mitochondrial function. Data mining demonstrated that NDRG4 was significantly down-regulated in PDAC tissues and cells. PDAC patients with low NDRG4 expression showed poor prognosis. Epigenetic regulation by DNA methylation was closely associated with NDRG4 down-regulation. NDRG4 overexpression dramatically suppressed PDAC cell growth and metastasis. Further functional analysis demonstrated that up-regulated NDRG4 in SW1990 and Canpan1 cells resulted in attenuated mitochondrial function, including reduced ATP production, decreased mitochondrial membrane potential, and increased fragmented mitochondria. However, opposite results were obtained for HPNE cells with NDRG4 knockdown. These results indicate that hypermethylation-driven silencing of NDRG4 can promote PDAC by regulating mitochondrial function and that NDRG4 could be as a potential biomarker for PDAC patients. [BMB Reports 2020; 53(12): 658-663]. more...
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- 2020
50. Striated Preferentially Expressed Protein Kinase (SPEG)-Deficient Skeletal Muscles Display Fewer Satellite Cells with Reduced Proliferation and Delayed Differentiation
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Tian Zhang, Shideh Kazerounian, Qifei Li, Jasmine Lin, Shiyu Luo, Samantha M. Rosen, and Pankaj B. Agrawal
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0301 basic medicine ,Population ,Cell ,Muscle Proteins ,chemistry.chemical_element ,Protein Serine-Threonine Kinases ,Calcium ,Biology ,Pathology and Forensic Medicine ,Myoblasts ,Mice ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Muscle, Skeletal ,education ,Protein kinase A ,Myosin-Light-Chain Kinase ,Gene ,Cell Proliferation ,education.field_of_study ,Skeletal muscle ,Regular Article ,Cell Differentiation ,Triad (anatomy) ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Apoptosis ,030217 neurology & neurosurgery ,Myopathies, Structural, Congenital - Abstract
Centronuclear myopathies (CNMs) are a subtype of congenital myopathies characterized by skeletal muscle weakness and an increase in the number of central myonuclei. SPEG (striated preferentially expressed protein kinase) has been identified as the sixth gene associated with CNM, and it has been shown that striated muscle-specific Speg-knockout (KO) mice have defective triad formation, abnormal excitation-contraction coupling, and calcium mishandling. The impact of SPEG deficiency on the survival and function of myogenic cells remains to be deciphered. In this study, the authors examined the overall population, proliferation, and differentiation of myogenic cells obtained from striated muscle-specific Speg-KO mice and compared them with wild-type (WT) controls. SPEG-deficient skeletal muscles contained fewer myogenic cells, which on further study demonstrated reduced proliferation and delayed differentiation compared with those from WT muscles. Regenerative response to skeletal muscle injury in Speg-KO mice was compared with that of WT mice, leading to the identification of similar abnormalities including fewer satellite cells, fewer dividing cells, and an increase in apoptotic cells in KO mice. Overall, these results reveal specific abnormalities in myogenic cell number and behavior associated with SPEG deficiency. Similar satellite cell defects have been reported in mouse models of MTM1- and DNM2-associated CNM, suggestive of shared underlying pathways. more...
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- 2020
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