Project description:Penile cancer (PeCa) is a relatively rare tumor entity but possesses higher morbidity and mortality rates especially in developing countries. To date, the concrete pathogenic signaling pathways and core machineries involved in tumorigenesis and progression of PeCa remain to be elucidated. Several studies suggested that miRNAs, which modulate gene expression at posttranscriptional level, were frequently mis-regulated and aberrantly expressed in human cancers. However, the miRNA profiles in human penile cancer have not been reported before. In this present study, the miRNA profiles were obtained from 10 fresh penile cancerous tissues and matched adjacent non-cancerous penile tissues via NGS. As a result, a total of 751 and 806 annotated miRNAs were identified in normal and cancerous penile tissues, respectively. Among which, 56 miRNAs with significantly different expression levels between paired penile tissues were identified. Subsequently, several annotated miRNAs were selected randomly and validated using quantitative real-time PCR. Compared with the previous publications regarding to the altered miRNAs expression in various types of cancers and especially genitourinary (prostate, bladder, kidney, testis) cancers, the most majority of deregulated miRNAs showed the similar expression pattern in penile cancer. Moreover, the bioinformatics analysis suggested that the putative target genes of the differentially expressed miRNAs were tightly associated with cell junction, proliferation, growth as well as genomic instability and so on, by modulating Wnt, MAPK, p53, PI3K-Akt, Notch, Hedgehog and TGF-β signaling pathways, which were all well-established to be involved in cancer initiation and progression. Our work presents a global view of the differentially expressed miRNAs and potentially regulatory networks of their target genes for clarifing the pathogenic transformation of normal penis to PeCa, which research resource also provides new insights into future investigations aimed to explore the in-depth mechanisms of miRNAs and other small RNAs including piRNAs in penile carcinogenesis regulation and effective target-specific theragnosis.
Project description:Penile cancer (PeCa) is a relatively rare tumor entity but possesses higher morbidity and mortality rates especially in developing countries. To date, the concrete pathogenic signaling pathways and core machineries involved in tumorigenesis and progression of PeCa remain to be elucidated. Several studies suggested miRNAs, which modulate gene expression at posttranscriptional level, were frequently mis-regulated and aberrantly expressed in human cancers. However, the miRNA profile in human PeCa has not been reported before. In this present study, the miRNA profile was obtained from 10 fresh penile cancerous tissues and matched adjacent non-cancerous tissues via next-generation sequencing. As a result, a total of 751 and 806 annotated miRNAs were identified in normal and cancerous penile tissues, respectively. Among which, 56 miRNAs with significantly different expression levels between paired tissues were identified. Subsequently, several annotated miRNAs were selected randomly and validated using quantitative real-time PCR. Compared with the previous publications regarding to the altered miRNAs expression in various cancers and especially genitourinary (prostate, bladder, kidney, testis) cancers, the most majority of deregulated miRNAs showed the similar expression pattern in penile cancer. Moreover, the bioinformatics analyses suggested that the putative target genes of differentially expressed miRNAs between cancerous and matched normal penile tissues were tightly associated with cell junction, proliferation, growth as well as genomic instability and so on, by modulating Wnt, MAPK, p53, PI3K-Akt, Notch and TGF-β signaling pathways, which were all well-established to participate in cancer initiation and progression. Our work presents a global view of the differentially expressed miRNAs and potentially regulatory networks of their target genes for clarifying the pathogenic transformation of normal penis to PeCa, which research resource also provides new insights into future investigations aimed to explore the in-depth mechanisms of miRNAs and other small RNAs including piRNAs in penile carcinogenesis regulation and effective target-specific theragnosis.
Project description:We aim to detect differential gene expression using RNA-seq between normal penile epithelial tissue and PB-Cre+ Smad4L/L ApcL/L penile tumors of mice
Project description:Purpose: Molecular mechanisms of penile corpus cavernosum aging and male age-related erectile dysfunction (ED) remain unclear. Here we profiled young and old rat penile corpus cavernousm by single-cell RNA sequencing (scRNA-seq). Methods:To map the single-cell transcriptomic landscape of penile corpus cavernosum during aging, we performed uniform manifold approximation and projection (UMAP), differential gene expression analysis (DGEs), pseudotime analysis and single-cell entropy algorithm to dissect cellular composition and transcriptional heterogeneity. For validation analysis, we further performed immunofluorescence studies on key molecules involved during penile corpus cavernosum aging. Results: After stringent filtering,transcriptomes of 14,879 single cells (8,557 young and 6,322 old) derived from penile corpus cavernosum of 5 young (3 months) and 5 old (23 months) rats were analyzed subsequently. Clustering analysis of cell-type specific gene expression identified 19 cell types, such as smooth muscle cells, endothelial cells, fibroblasts,myofibroblasts and immune cells.Transcriptomic analyses revealed that transcriptional alterations across all cell types exhibited distinct properties rather than universally consistent. DGEs analysis demonstrated that genes related to extracellular matrix organization were highly expressed. Among these cell types, fibroblasts showed apparent heterogeneities. By performing pseudotime and single-cell entropy analysis on fibroblasts, we observed the age-associated decrease of entropy, and aged fibroblasts were found to adopt senescent secretory phenotype, as evidenced by the high expression of genes associated with the senescence-associated secretory phenotype (SASP). Since eliminating senescent cells or SASP were demonstrated to improve health and life span, we further investigated the distinct senescence-related gene expression signatures across all cell types during aging. Conclusions: We plotted a cellular atlas of penile corpus cavernosum, and revealed the molecular alterations of aging cells, especially fibroblasts. Our work will deepen the understanding of the heterogeneity among certain cell types during penile corpus cavernosum aging and provide novel entry points for the age-associated ED treatment.
Project description:The circadian clock drives daily rhythms of gene expression and physiological functions across tissues throughout the body. Advances in next-generation DNA sequencing have provided extensive insights into gene expression at the RNA level, but more functional information at the protein level with sufficient depth has been limited by technical challenges. Recently, the next-generation mass spectrometer Orbitrap Astral was developed, allowing us to quantify protein abundance with greater sensitivity and accuracy. In this study, we generated a comprehensive mouse circadian proteome atlas (available in URL) by analyzing protein abundance in 32 tissues including the suprachiasmatic nucleus (SCN) across the day using Orbitrap Astral. Data-independent acquisition of 500 samples including developmental samples revealed the spatiotemporal profiles of 18,751 proteins, accounting for 73% of all proteins registered in UniProt. Proteome and phospho-proteome analyses of whole-cell and nuclear proteins in the liver uncovered circadian profiles not only in protein abundance but also in subcellular localization and post-translational modification. Notably, proteome analysis revealed global changes in protein phosphorylation status in hPER2-S662G mutant mice, a genetic model of human familial advanced sleep phase (FASP). This multi-tissue circadian proteome atlas provides a fundamental resource for understanding when, where, and which proteins are expressed and function.
Project description:The circadian clock drives daily rhythms of gene expression and physiological functions across tissues throughout the body. Advances in next-generation DNA sequencing have provided extensive insights into gene expression at the RNA level, but more functional information at the protein level with sufficient depth has been limited by technical challenges. Recently, the next-generation mass spectrometer Orbitrap Astral was developed, allowing us to quantify protein abundance with greater sensitivity and accuracy. In this study, we generated a comprehensive mouse circadian proteome atlas (available in URL) by analyzing protein abundance in 32 tissues including the suprachiasmatic nucleus (SCN) across the day using Orbitrap Astral. Data-independent acquisition of 500 samples including developmental samples revealed the spatiotemporal profiles of 18,751 proteins, accounting for 73% of all proteins registered in UniProt. Proteome and phospho-proteome analyses of whole-cell and nuclear proteins in the liver uncovered circadian profiles not only in protein abundance but also in subcellular localization and post-translational modification. Notably, proteome analysis revealed global changes in protein phosphorylation status in hPER2-S662G mutant mice, a genetic model of human familial advanced sleep phase (FASP). This multi-tissue circadian proteome atlas provides a fundamental resource for understanding when, where, and which proteins are expressed and function.
Project description:The circadian clock drives daily rhythms of gene expression and physiological functions across tissues throughout the body. Advances in next-generation DNA sequencing have provided extensive insights into gene expression at the RNA level, but more functional information at the protein level with sufficient depth has been limited by technical challenges. Recently, the next-generation mass spectrometer Orbitrap Astral was developed, allowing us to quantify protein abundance with greater sensitivity and accuracy. In this study, we generated a comprehensive mouse circadian proteome atlas (available in URL) by analyzing protein abundance in 32 tissues including the suprachiasmatic nucleus (SCN) across the day using Orbitrap Astral. Data-independent acquisition of 500 samples including developmental samples revealed the spatiotemporal profiles of 18,751 proteins, accounting for 73% of all proteins registered in UniProt. Proteome and phospho-proteome analyses of whole-cell and nuclear proteins in the liver uncovered circadian profiles not only in protein abundance but also in subcellular localization and post-translational modification. Notably, proteome analysis revealed global changes in protein phosphorylation status in hPER2-S662G mutant mice, a genetic model of human familial advanced sleep phase (FASP). This multi-tissue circadian proteome atlas provides a fundamental resource for understanding when, where, and which proteins are expressed and function.
Project description:The circadian clock drives daily rhythms of gene expression and physiological functions across tissues throughout the body. Advances in next-generation DNA sequencing have provided extensive insights into gene expression at the RNA level, but more functional information at the protein level with sufficient depth has been limited by technical challenges. Recently, the next-generation mass spectrometer Orbitrap Astral was developed, allowing us to quantify protein abundance with greater sensitivity and accuracy. In this study, we generated a comprehensive mouse circadian proteome atlas (available in https://chiba1.dynacom.co.jp/mcp_atlas/) by analyzing protein abundance in 32 tissues including the suprachiasmatic nucleus (SCN) across the day using Orbitrap Astral. Data-independent acquisition of 584 samples including developmental samples revealed the spatiotemporal profiles of 18,956 proteins, accounting for 73.8% of all proteins registered in UniProt. Proteome and phospho-proteome analyses of whole-cell and nuclear proteins in the liver uncovered circadian profiles not only in protein abundance but also in subcellular localization and post-translational modification. Notably, proteome analysis revealed global changes in protein phosphorylation status in hPER2-S662G mutant mice, a genetic model of human familial advanced sleep phase (FASP). This multi-tissue circadian proteome atlas provides a fundamental resource for understanding when, where, and which proteins are expressed and function.
Project description:The circadian clock drives daily rhythms of gene expression and physiological functions across tissues throughout the body. Advances in next-generation DNA sequencing have provided extensive insights into gene expression at the RNA level, but more functional information at the protein level with sufficient depth has been limited by technical challenges. Recently, the next-generation mass spectrometer Orbitrap Astral was developed, allowing us to quantify protein abundance with greater sensitivity and accuracy. In this study, we generated a comprehensive mouse circadian proteome atlas (available in URL) by analyzing protein abundance in 32 tissues including the suprachiasmatic nucleus (SCN) across the day using Orbitrap Astral. Data-independent acquisition of 500 samples including developmental samples revealed the spatiotemporal profiles of 18,751 proteins, accounting for 73% of all proteins registered in UniProt. Proteome and phospho-proteome analyses of whole-cell and nuclear proteins in the liver uncovered circadian profiles not only in protein abundance but also in subcellular localization and post-translational modification. Notably, proteome analysis revealed global changes in protein phosphorylation status in hPER2-S662G mutant mice, a genetic model of human familial advanced sleep phase (FASP). This multi-tissue circadian proteome atlas provides a fundamental resource for understanding when, where, and which proteins are expressed and function.
Project description:The circadian clock drives daily rhythms of gene expression and physiological functions across tissues throughout the body. Advances in next-generation DNA sequencing have provided extensive insights into gene expression at the RNA level, but more functional information at the protein level with sufficient depth has been limited by technical challenges. Recently, the next-generation mass spectrometer Orbitrap Astral was developed, allowing us to quantify protein abundance with greater sensitivity and accuracy. In this study, we generated a comprehensive mouse circadian proteome atlas (available in URL) by analyzing protein abundance in 32 tissues including the suprachiasmatic nucleus (SCN) across the day using Orbitrap Astral. Data-independent acquisition of 500 samples including developmental samples revealed the spatiotemporal profiles of 18,751 proteins, accounting for 73% of all proteins registered in UniProt. Proteome and phospho-proteome analyses of whole-cell and nuclear proteins in the liver uncovered circadian profiles not only in protein abundance but also in subcellular localization and post-translational modification. Notably, proteome analysis revealed global changes in protein phosphorylation status in hPER2-S662G mutant mice, a genetic model of human familial advanced sleep phase (FASP). This multi-tissue circadian proteome atlas provides a fundamental resource for understanding when, where, and which proteins are expressed and function.