{"title":"Elimination of physiological senescent cutaneous cells in a novel p16‐dependent senolytic mouse model impacts lipid metabolism in skin aging","authors":"Yuma Sugiyama, Yoichiro Kawabe, Tanenobu Harada, Yu Aoki, Keiko Tsuji, Daijiro Sugiyama, Mitsuo Maruyama","doi":"10.1111/gtc.13163","DOIUrl":"https://doi.org/10.1111/gtc.13163","url":null,"abstract":"The evidence of the correlation between cellular senescence and aging has increased in research with animal models. These models have been intentionally generated to target and regulate cellular senescent cells with the promoter activity of <jats:italic>p16</jats:italic><jats:sup><jats:italic>Ink4a</jats:italic></jats:sup> or <jats:italic>p19</jats:italic><jats:sup><jats:italic>Arf</jats:italic></jats:sup>, genes that are highly expressed in aging cells. However, the senolytic efficiency in various organs and cells from these models represents unexpected variation and diversity in some cases. We have generated a novel knock‐in model, <jats:italic>p16</jats:italic>tdT‐hDTR mice, which possess tdTomato and human diphtheria toxin receptor (hDTR) downstream of <jats:italic>Cdkn2a</jats:italic>, an endogenous <jats:italic>p16</jats:italic><jats:sup><jats:italic>Ink4a</jats:italic></jats:sup> gene. We successfully demonstrated that p16‐derived tdTomato and hDTR expressions are observed in these mouse embryo fibroblasts and following treatment with diphtheria toxin (DT) eliminates those cells. Furthermore, we demonstrated the efficacy of eliminating p16‐positive cells in vivo, and also observed a tendency to decrease their cutaneous SA‐β‐gal activity after subcutaneous DT injection into <jats:italic>p16</jats:italic>tdT‐hDTR mice. In particular, comprehensive gene expression analysis in skin revealed that upregulated genes related to lipid metabolisms with aging exhibited remarkable expressions under the senolysis. These results clearly unveiled p16‐positive senescent cells contribute to age‐related changes in skin.","PeriodicalId":12742,"journal":{"name":"Genes to Cells","volume":null,"pages":null},"PeriodicalIF":2.1,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253933","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Neonatal Fc receptor is a functional receptor for classical human astrovirus","authors":"Kei Haga, Takashi Tokui, Kana Miyamoto, Reiko Takai‐Todaka, Shiori Kudo, Azusa Ishikawa, Ryoka Ishiyama, Akiko Kato, Masaru Yokoyama, Kazuhiko Katayama, Akira Nakanishi","doi":"10.1111/gtc.13160","DOIUrl":"https://doi.org/10.1111/gtc.13160","url":null,"abstract":"Human astrovirus (HAstV) is a global cause of gastroenteritis in infants, the elderly, and the immunocompromised. However, the molecular mechanisms that control its susceptibility are not fully understood, as the functional receptor used by the virus has yet to be identified. Here, a genome‐wide CRISPR‐Cas9 library screen in Caco2 cells revealed that the neonatal Fc receptor (FcRn) can function as a receptor for classical HAstV (<jats:italic>Mamastrovirus</jats:italic> genotype 1). Deletion of <jats:italic>FCGRT</jats:italic> or <jats:italic>B2M</jats:italic>, which encode subunits of FcRn, rendered Caco2 cells and intestinal organoid cells resistant to HAstV infection. We also showed that human FcRn expression renders non‐susceptible cells permissive to viral infection and that FcRn binds directly to the HAstV spike protein. Therefore, our findings provide insight into the entry mechanism of HAstV into susceptible cells. We anticipate that this information can be used to develop new therapies targeting human astroviruses, providing new strategies to treat this global health issue.","PeriodicalId":12742,"journal":{"name":"Genes to Cells","volume":null,"pages":null},"PeriodicalIF":2.1,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Capsaicin modulates TRPV1, induces β‐defensin expression, and regulates NF‐κB in oral senescent cells and a murine model","authors":"Yoriko Ikuyo, Haruna Yokoi, Jingshu Wang, Masae Furukawa, Resmi Raju, Mitsuyoshi Yamada, Yu Aoki, Kenji Matsushita","doi":"10.1111/gtc.13158","DOIUrl":"https://doi.org/10.1111/gtc.13158","url":null,"abstract":"Aging is associated with a decline in oral immune function, marked by reduced levels of antimicrobial peptides such as defensins. Capsaicin, a bioactive component found in chili peppers, has been theorized to modulate immune responses through specific receptor pathways. This study examined the effects of aging on oral defensin levels and the potential mitigating role of capsaicin, mediated by the immune response in oral tissues. We conducted a comparative analysis between young and aged mice, with or without capsaicin supplementation, for 3 months. The effect of capsaicin was also studied in vitro in senescence‐induced human oral keratinocytes. We found that aging did not reduce defensin levels uniformly but did so in some instances. Capsaicin treatment increased defensin levels in these cases, potentially through transient receptor potential cation channel subfamily V member 1 (TRPV1)‐mediated pathways in the oral cavity. Capsaicin supplementation may counteract age‐related declines in oral defensin levels, enabling the maintenance of oral immune function during aging.","PeriodicalId":12742,"journal":{"name":"Genes to Cells","volume":null,"pages":null},"PeriodicalIF":2.1,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142253934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Genes to CellsPub Date : 2024-09-12DOI: 10.1111/gtc.13159
Hirotsugu Hino, Kaori Takaki, Mika Kobe, Satoru Mochida
{"title":"Development of luminescent probes for real‐time detection of the CDK/PP2A balance during the cell cycle","authors":"Hirotsugu Hino, Kaori Takaki, Mika Kobe, Satoru Mochida","doi":"10.1111/gtc.13159","DOIUrl":"https://doi.org/10.1111/gtc.13159","url":null,"abstract":"From a biochemical viewpoint, the cell cycle is controlled by the phosphorylation of cyclin‐dependent kinase (CDK) substrates, and the phosphorylation level is determined by the enzymatic balance between CDK and protein phosphatase 2A (PP2A). However, the conventional techniques for analyzing protein phosphorylation using radioisotopes and antibodies involve many operational steps and take days before obtaining results, making them difficult to apply to high‐throughput screening and real‐time observations. In this study, we developed luminescent probes with a light intensity that changes depending on its phosphorylation state. We modified the Nano‐lantern probe (<jats:italic>Renilla</jats:italic> luciferase‐based Ca<jats:sup>2+</jats:sup> probe) by introducing a CDK‐substrate peptide and a phosphopeptide‐binding domain into the luciferase. Our initial trial resulted in new probes that could report the CDK/PP2A balance in a purified system. Further modifications of these probes (replacing the phospho‐Ser with phospho‐Thr and randomly replacing its surrounding amino acids) improved the dynamic range by up to four‐fold, making them practical for use in the <jats:italic>Xenopus</jats:italic> egg extracts system, where many physiological events can be reproduced. Taken together, our new probes enabled the monitoring of the CDK/PP2A balance in real time, and are applicable to high‐throughput systems; the new probes thus appear promising for use in substrate and drug screening.","PeriodicalId":12742,"journal":{"name":"Genes to Cells","volume":null,"pages":null},"PeriodicalIF":2.1,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142189055","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"RNA–DNA hybrids on protein coding genes are stabilized by loss of RNase H and are associated with DNA damages during S‐phase in fission yeast","authors":"Tomoko Sagi, Daichi Sadato, Kazuto Takayasu, Hiroyuki Sasanuma, Yutaka Kanoh, Hisao Masai","doi":"10.1111/gtc.13157","DOIUrl":"https://doi.org/10.1111/gtc.13157","url":null,"abstract":"RNA–DNA hybrid is a part of the R‐loop which is an important non‐standard nucleic acid structure. RNA–DNA hybrid/R‐loop causes genomic instability by inducing DNA damages or inhibiting DNA replication. It also plays biologically important roles in regulation of transcription, replication, recombination and repair. Here, we have employed catalytically inactive human RNase H1 mutant (D145N) to visualize RNA–DNA hybrids and map their genomic locations in fission yeast cells. The RNA–DNA hybrids appear as multiple nuclear foci in <jats:italic>rnh1∆rnh201∆</jats:italic> cells lacking cellular RNase H activity, but not in the wild‐type. The majority of RNA–DNA hybrid loci are detected at the protein coding regions and tRNA. In <jats:italic>rnh1∆rnh201∆</jats:italic> cells, cells with multiple Rad52 foci increase during S‐phase and about 20% of the RNA–DNA hybrids overlap with Rad52 loci. During S‐phase, more robust association of Rad52 with RNA–DNA hybrids was observed in the protein coding region than in M‐phase. These results suggest that persistent RNA–DNA hybrids in the protein coding region in <jats:italic>rnh1∆rnh201∆</jats:italic> cells generate DNA damages during S‐phase, potentially through collision with DNA replication forks.","PeriodicalId":12742,"journal":{"name":"Genes to Cells","volume":null,"pages":null},"PeriodicalIF":2.1,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142189057","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Identification of genes contributing to attenuation of rat model of galactose-induced cataract by pyruvate","authors":"Fuuga Masuda, Mayumi Inami, Yoshihiro Takamura, Masaru Inatani, Masaya Oki","doi":"10.1111/gtc.13150","DOIUrl":"10.1111/gtc.13150","url":null,"abstract":"<p>Cataracts are a disease that reduces vision due to opacity formation of the lens. Diabetic cataracts occur at young age and progress relatively quickly, so the development of effective treatment has been awaited. Several studies have shown that pyruvate inhibits oxidative stress and glycation of lens proteins, which contribute to onset of diabetic cataracts. However, detailed molecular mechanisms have not been revealed. In this study, we attempted to reduce galactose-induced opacity by pyruvate with rat ex vivo model. Rat lenses were extracted and cultured in galactose-containing medium to induce lens opacity. After opacity had developed, continued culturing with pyruvate in the medium resulted in a reduction of lens opacity. Subsequently, we conducted microarray analysis to investigate the genes that contribute to the therapeutic effect. We performed quantitative expression measurements using RT-qPCR for extracted genes that were upregulated in cataract-induced lenses and downregulated in pyruvate-treated lenses, resulting in the identification of 34 candidate genes. Functional analysis using the STRING database suggests that metallothionein-related factors (<i>Mt1a</i>, <i>Mt1m</i>, and <i>Mt2A</i>) and epithelial-mesenchymal transition-related factors (<i>Acta2</i>, <i>Anxa1</i>, <i>Cd81</i>, <i>Mki67</i>, <i>Timp1</i>, and <i>Tyms</i>) contribute to the therapeutic effect of cataracts.</p>","PeriodicalId":12742,"journal":{"name":"Genes to Cells","volume":null,"pages":null},"PeriodicalIF":1.3,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/gtc.13150","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142106698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"L3MBTL2 maintains MYCN-amplified neuroblastoma cell proliferation through silencing NRIP3 and BRME1 genes","authors":"Ryu Okada, Hisanori Takenobu, Shunpei Satoh, Ryuichi P. Sugino, Ritsuko Onuki, Masayuki Haruta, Kyosuke Mukae, Atsuko Nakazawa, Jesmin Akter, Miki Ohira, Takehiko Kamijo","doi":"10.1111/gtc.13148","DOIUrl":"10.1111/gtc.13148","url":null,"abstract":"<p>Epigenetic alterations critically affect tumor development. Polycomb-group complexes constitute an evolutionarily conserved epigenetic machinery that regulates stem cell fate and development. They are implicated in tumorigenesis, primarily via histone modification. Polycomb repressive complex 1 (PRC1) complexes 1–6 (PRC1.1–6) mediate the ubiquitination of histone H2A on lysine 119 (H2AK119ub). Here, we studied the functional roles of a PRC1.6 molecule, L3MBTL2, in neuroblastoma (NB) cells. <i>L3MBTL2</i>-knockout and knockdown revealed that L3MBTL2 depletion suppressed NB cell proliferation via cell-cycle arrest and gamma-H2A.X upregulation. <i>L3MBTL2-</i>knockout profoundly suppressed xenograft tumor formation. Transcriptome analysis revealed suppressed cell-cycle-related and activated differentiation-related pathways. <i>Break repair meiotic recombinase recruitment factor 1</i> (<i>BRME1</i>) and <i>nuclear receptor interacting protein 3</i> (<i>NRIP3</i>) were notably de-repressed by <i>L3MBTL2</i>-knockout. The deletion of <i>L3MBTL2</i> reduced enrichment of H2AK119ub and PCGF6 at transcriptional start site proximal regions of the targets. Add-back studies unveiled the importance of L3MBTL2-BRME1 and -NRIP3 axes for NB cell proliferation. We further manifested the association of MYCN with de-repression of <i>NRIP3</i> in an <i>L3MBTL2</i>-deficient context. Therefore, this study first revealed the significance of <i>L3MBTL2-</i>mediated gene silencing in MYCN-amplified NB cells.</p>","PeriodicalId":12742,"journal":{"name":"Genes to Cells","volume":null,"pages":null},"PeriodicalIF":1.3,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142072565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Reconstruction of artificial nuclei with nuclear import activity in living mouse oocytes","authors":"Nao Yonezawa, Tomoko Shindo, Haruka Oda, Hiroshi Kimura, Yasushi Hiraoka, Tokuko Haraguchi, Kazuo Yamagata","doi":"10.1111/gtc.13149","DOIUrl":"10.1111/gtc.13149","url":null,"abstract":"<p>In eukaryotes, DNA is housed within the cell nucleus. Molecules required for the formation of a nucleus have been identified using in vitro systems with frog egg extracts and in vivo imaging of somatic cells. However, little is known about the physicochemical factors and conditions required for nuclear formation in mouse oocytes. In this study, using a reconstitution approach with purified DNA, we aimed to determine factors, such as the amount and timing of DNA introduction, required for the formation of nuclei with nuclear transport activity in mouse oocytes. T4 phage DNA (~166 kbp) was microinjected into strontium-activated oocytes to evaluate the conditions appropriate for nuclear formation. Microinjection of 100–500 ng/μL of T4 DNA, but not 20 ng/μL, was sufficient for the formation of nucleus-like structures. Furthermore, microinjection of DNA during metaphase II to telophase II, but not during interphase, was sufficient. Electron and fluorescence microscopy showed that T4 DNA-induced nucleus-like structures had nuclear lamina and nuclear pore complex structures similar to those of natural nuclei, as well as nuclear import activity. These results suggest that exogenous DNA can form artificial nuclei with nuclear transport functions in mouse oocytes, regardless of the sequence or source of the DNA.</p>","PeriodicalId":12742,"journal":{"name":"Genes to Cells","volume":null,"pages":null},"PeriodicalIF":1.3,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141975533","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Genes to CellsPub Date : 2024-08-13DOI: 10.1111/gtc.13151
Mifra Faiz, Maggie L. Kalev-Zylinska, Caitlin Dunstan-Harrison, Dean C. Singleton, Michael P. Hay, Elizabeth C. Ledgerwood
{"title":"Megakaryocyte maturation involves activation of the adaptive unfolded protein response","authors":"Mifra Faiz, Maggie L. Kalev-Zylinska, Caitlin Dunstan-Harrison, Dean C. Singleton, Michael P. Hay, Elizabeth C. Ledgerwood","doi":"10.1111/gtc.13151","DOIUrl":"10.1111/gtc.13151","url":null,"abstract":"<p>Endoplasmic reticulum stress triggers the unfolded protein response (UPR) to promote cell survival or apoptosis. Transient endoplasmic reticulum stress activation has been reported to trigger megakaryocyte production, and UPR activation has been reported as a feature of megakaryocytic cancers. However, the role of UPR signaling in megakaryocyte biology is not fully understood. We studied the involvement of UPR in human megakaryocytic differentiation using PMA (phorbol 12-myristate 13-acetate)-induced maturation of megakaryoblastic cell lines and thrombopoietin-induced differentiation of human peripheral blood-derived progenitors. Our results demonstrate that an adaptive UPR is a feature of megakaryocytic differentiation and that this response is not associated with ER stress-induced apoptosis. Differentiation did not alter the response to the canonical endoplasmic reticulum stressors DTT or thapsigargin. However, thapsigargin, but not DTT, inhibited differentiation, consistent with the involvement of Ca<sup>2+</sup> signaling in megakaryocyte differentiation.</p>","PeriodicalId":12742,"journal":{"name":"Genes to Cells","volume":null,"pages":null},"PeriodicalIF":1.3,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/gtc.13151","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141975532","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Meflin/ISLR is a marker of adipose stem and progenitor cells in mice and humans that suppresses white adipose tissue remodeling and fibrosis","authors":"Toshikazu Ishihara, Katsuhiro Kato, Kotaro Matsumoto, Miyako Tanaka, Akitoshi Hara, Yukihiro Shiraki, Hidenori Morisaki, Yuya Urano, Ryota Ando, Kisuke Ito, Shinji Mii, Nobutoshi Esaki, Kazuhiro Furuhashi, Mikito Takefuji, Takayoshi Suganami, Toyoaki Murohara, Atsushi Enomoto","doi":"10.1111/gtc.13154","DOIUrl":"10.1111/gtc.13154","url":null,"abstract":"<p>Identifying specific markers of adipose stem and progenitor cells (ASPCs) in vivo is crucial for understanding the biology of white adipose tissues (WAT). PDGFRα-positive perivascular stromal cells represent the best candidates for ASPCs. This cell lineage differentiates into myofibroblasts that contribute to the impairment of WAT function. However, ASPC marker protein(s) that are functionally crucial for maintaining WAT homeostasis are unknown. We previously identified Meflin as a marker of mesenchymal stem cells (MSCs) in bone marrow and tissue-resident perivascular fibroblasts in various tissues. We also demonstrated that Meflin maintains the undifferentiated status of MSCs/fibroblasts. Here, we show that Meflin is expressed in WAT ASPCs. A lineage-tracing experiment showed that Meflin<sup>+</sup> ASPCs proliferate in the WAT of obese mice induced by a high-fat diet (HFD), while some of them differentiate into myofibroblasts or mature adipocytes. Meflin knockout mice fed an HFD exhibited a significant fibrotic response as well as increases in adipocyte cell size and the number of crown-like structures in WAT, accompanied by impaired glucose tolerance. These data suggested that Meflin expressed by ASPCs may have a role in reducing disease progression associated with WAT dysfunction.</p>","PeriodicalId":12742,"journal":{"name":"Genes to Cells","volume":null,"pages":null},"PeriodicalIF":1.3,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/gtc.13154","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141970991","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}