Lauren Belfiore, Anjali Balakrishnan, Yacine Touahri, Dawn Zinyk, Humna Noman, Satoshi Okawa, Jeff Biernaskie, Carol Schuurmans
{"title":"神经胶质转录因子组合表达诱导小鼠胚胎成纤维细胞雪旺细胞特异性基因表达。","authors":"Lauren Belfiore, Anjali Balakrishnan, Yacine Touahri, Dawn Zinyk, Humna Noman, Satoshi Okawa, Jeff Biernaskie, Carol Schuurmans","doi":"10.1002/dvdy.70054","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Schwann cells provide peripheral nerve trophic support, myelinate axons, and assist in repair. However, Schwann cell repair capacity is limited by chronic injury, disease, and aging. Schwann cell reprogramming is a cellular conversion strategy that could provide a renewable cell supply to repair injured nerves. Here, we developed a plasmid-based approach to test the Schwann cell conversion potential of four glial transcription factors.</p><p><strong>Results: </strong>We employed four transcription factors implicated in Schwann cell differentiation and repair: Sox10, Sox2, Jun, and Pax3. Expression vectors were generated for Sox10 alone and two triple transcription factor combinations: Jun-Pax3-Sox2 (triple 1, T1) and Sox10-Jun-Sox2 (triple 2, T2). Mouse embryonic fibroblasts (MEFs) were transfected with these vectors, transferred to glial inductive media, and Schwann cell-marker expression was in assessed by immunostaining, flow cytometry, and qPCR. All expression vectors repressed fibroblast-specific gene expression. However, T2 was most efficient at generating O4<sup>+</sup> Schwann cell-like cells, which had some capacity to myelinate denervated axons from explanted dorsal root ganglia. In comparison, T1 more efficiently induced repair Schwann cell-marker expression in converted O4<sup>+</sup> cells.</p><p><strong>Conclusions: </strong>T1 and T2 convert MEFs to Schwann cells with different efficacies and gene expression profiles, and may provide cell-based therapies for peripheral nerve repair.</p>","PeriodicalId":11247,"journal":{"name":"Developmental Dynamics","volume":" ","pages":""},"PeriodicalIF":1.5000,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Combinatorial expression of glial transcription factors induces Schwann cell-specific gene expression in mouse embryonic fibroblasts.\",\"authors\":\"Lauren Belfiore, Anjali Balakrishnan, Yacine Touahri, Dawn Zinyk, Humna Noman, Satoshi Okawa, Jeff Biernaskie, Carol Schuurmans\",\"doi\":\"10.1002/dvdy.70054\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Schwann cells provide peripheral nerve trophic support, myelinate axons, and assist in repair. However, Schwann cell repair capacity is limited by chronic injury, disease, and aging. Schwann cell reprogramming is a cellular conversion strategy that could provide a renewable cell supply to repair injured nerves. Here, we developed a plasmid-based approach to test the Schwann cell conversion potential of four glial transcription factors.</p><p><strong>Results: </strong>We employed four transcription factors implicated in Schwann cell differentiation and repair: Sox10, Sox2, Jun, and Pax3. Expression vectors were generated for Sox10 alone and two triple transcription factor combinations: Jun-Pax3-Sox2 (triple 1, T1) and Sox10-Jun-Sox2 (triple 2, T2). Mouse embryonic fibroblasts (MEFs) were transfected with these vectors, transferred to glial inductive media, and Schwann cell-marker expression was in assessed by immunostaining, flow cytometry, and qPCR. All expression vectors repressed fibroblast-specific gene expression. However, T2 was most efficient at generating O4<sup>+</sup> Schwann cell-like cells, which had some capacity to myelinate denervated axons from explanted dorsal root ganglia. In comparison, T1 more efficiently induced repair Schwann cell-marker expression in converted O4<sup>+</sup> cells.</p><p><strong>Conclusions: </strong>T1 and T2 convert MEFs to Schwann cells with different efficacies and gene expression profiles, and may provide cell-based therapies for peripheral nerve repair.</p>\",\"PeriodicalId\":11247,\"journal\":{\"name\":\"Developmental Dynamics\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":1.5000,\"publicationDate\":\"2025-06-19\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Developmental Dynamics\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1002/dvdy.70054\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"ANATOMY & MORPHOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Developmental Dynamics","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1002/dvdy.70054","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"ANATOMY & MORPHOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
背景:雪旺细胞提供周围神经营养支持,髓鞘轴突,并协助修复。然而,雪旺细胞的修复能力受到慢性损伤、疾病和衰老的限制。雪旺细胞重编程是一种细胞转化策略,可以提供可再生的细胞供应来修复受损的神经。在这里,我们开发了一种基于质粒的方法来测试四种胶质转录因子的雪旺细胞转化潜力。结果:我们使用了四个参与雪旺细胞分化和修复的转录因子:Sox10、Sox2、Jun和Pax3。生成单独Sox10和两个三重转录因子组合Jun-Pax3-Sox2 (triple 1, T1)和Sox10- jun - sox2 (triple 2, T2)的表达载体。将这些载体转染小鼠胚胎成纤维细胞(mef),转移到胶质诱导培养基上,通过免疫染色、流式细胞术和qPCR评估雪旺细胞标志物的表达。所有表达载体均抑制成纤维细胞特异性基因的表达。然而,T2最有效的是产生O4+雪旺细胞样细胞,这些细胞具有一定的能力,能够从外植的背根神经节中形成髓鞘。相比之下,T1在转化的O4+细胞中更有效地诱导修复雪旺细胞标志物的表达。结论:T1和T2将mef转化为雪旺细胞的效果和基因表达谱不同,可能为周围神经修复提供细胞基础疗法。
Combinatorial expression of glial transcription factors induces Schwann cell-specific gene expression in mouse embryonic fibroblasts.
Background: Schwann cells provide peripheral nerve trophic support, myelinate axons, and assist in repair. However, Schwann cell repair capacity is limited by chronic injury, disease, and aging. Schwann cell reprogramming is a cellular conversion strategy that could provide a renewable cell supply to repair injured nerves. Here, we developed a plasmid-based approach to test the Schwann cell conversion potential of four glial transcription factors.
Results: We employed four transcription factors implicated in Schwann cell differentiation and repair: Sox10, Sox2, Jun, and Pax3. Expression vectors were generated for Sox10 alone and two triple transcription factor combinations: Jun-Pax3-Sox2 (triple 1, T1) and Sox10-Jun-Sox2 (triple 2, T2). Mouse embryonic fibroblasts (MEFs) were transfected with these vectors, transferred to glial inductive media, and Schwann cell-marker expression was in assessed by immunostaining, flow cytometry, and qPCR. All expression vectors repressed fibroblast-specific gene expression. However, T2 was most efficient at generating O4+ Schwann cell-like cells, which had some capacity to myelinate denervated axons from explanted dorsal root ganglia. In comparison, T1 more efficiently induced repair Schwann cell-marker expression in converted O4+ cells.
Conclusions: T1 and T2 convert MEFs to Schwann cells with different efficacies and gene expression profiles, and may provide cell-based therapies for peripheral nerve repair.
期刊介绍:
Developmental Dynamics, is an official publication of the American Association for Anatomy. This peer reviewed journal provides an international forum for publishing novel discoveries, using any model system, that advances our understanding of development, morphology, form and function, evolution, disease, stem cells, repair and regeneration.