Engineering of functional smooth muscle cells from gastrointestinal myofibroblast

胃肠道肌成纤维细胞的功能性平滑肌细胞工程

• 批准号: 8888878
• 负责人: Seungil Ro
• 金额: $ 32.77万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-15 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8888878
• 关键词: Adopted; Animal Model; Automobile Driving; Back; Binding; Blood Vessels; Boxing; Calcium; Cell Line; Cell Separation; Cell Therapy; Cells; Clinic; Clinical; Colon; Complex; Consult; CpG Islands; DNA Modification Methylases; Development; Disease; Doctor of Philosophy; Ectopic Expression; Embryo; Engineering; Epigenetic Process; Fibrosis; Functional disorder; Gastrointestinal Motility; Gastrointestinal tract structure; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Genetic Engineering; Genetic Identity; Growth Factor; Human; Hyperplasia; Hypertrophy; Image; Injury; Interstitial Cell of Cajal; Intestinal Obstruction; Intestines; Knowledge; Label; LacZ Genes; Literature; Maps; Methods; Methylation; Modeling; Molecular; Molecular Genetics; Molecular Profiling; Moon; Mus; Myofibroblast; Names; Neurons; Organ; Phenotype; Platelet-Derived Growth Factor alpha Receptor; Postdoctoral Fellow; Preventive Medicine; Principal Investigator; Process; Proliferating; Recovery; Regenerative Medicine; Research; Serum Response Factor; Small Intestines; Smooth Muscle; Smooth Muscle Myocytes; System; Techniques; Testing; Tissues; adult stem cell; cell dedifferentiation; cell transformation; cell type; cellular engineering; clinical application; cofactor; gastrointestinal; genome-wide; jejunum; motility disorder; myocardin; programs; public health relevance; repaired; response; tissue regeneration; tool; transcription factor

 DESCRIPTION (provided by applicant): Dysfunctions of smooth muscle cells (SMCs) are associated with hypertrophy, hyperplasia and fibrosis found in SM diseases including gastrointestinal (GI) bowel obstruction and Hirschprung's disease. Defective SMCs are dedifferentiated cells that have lost their contractile phenotype. The phenotypic change of SMCs to non- functional dedifferentiated cells has been discovered in vascular injuries ~50 years ago, but the phenotypic and genetic identity of the transformed cells are still unknown. We have found that the transformed cells in the hypertrophic small intestine are myofibroblast like cells that weakly express platelet-derived growth factor receptor alpha and beta (PDGFRa/ß) (called "PDGFRalowßhigh cells"). We have isolated differentiated PDGFRahigh cells and SMCs from colon and jejunum, and obtained genome scale gene expression profiles. By analyzing the transcriptomes, we identified SMC-specific transcription factors including SRF and MYOCD. Moreover, we found that the phenotype of SMCs is regulated by SRF and an epigenetic modulator DNA methyltransferase 1 and 3a (DNMT1/3a). The present project seeks to uncover a cellar and genetic mechanism for understanding how SMCs are dedifferentiated during the development of intestinal SM hypertrophy and how dedifferentiated cells are redifferentiated into SMCs during the recovery process. Furthermore, this project aims to isolate dedifferentiated cells, establish a cell line in culture, and genetically engineer SMCs from the cell line using SMC-specific transcription factors and the epigenetic regulator DNMT1/3a. To achieve these aims, we have formed a collaborative, multi-institutional research team with Drs. Seungil Ro (PhD, PI), Moon young Lee (MD/PhD, studying the plasticity of SMCs in the hypertrophic condition), a to-be-hired postdoc (PhD, studying the engineering of SMCs from PDGRFalow cells), Douglas Redelman (PhD, cell isolation), Gabsang Lee (PhD/DVM, cell line establishment), Terence Smith (PhD, calcium imaging), Laren Becker (MD/PhD, tissue supply and consulting), Kent Sasse (MD, tissue supply), and Joseph Miano and Kenton Sanders (PhDs, research tool supply and project consulting). These studies will significantly advance our understanding on the phenotypic transition of SMCs at the transcriptome level and could offer a human SMC line for clinical use to repair damaged or non-functional SMCs found in GI SM diseases.

 描述（由申请方提供）：平滑肌细胞（SMC）功能障碍与SM疾病（包括胃肠道（GI）肠梗阻和Hirschprung病）中发现的肥大、增生和纤维化相关。有缺陷的SMC是已经失去其收缩表型的去分化细胞。平滑肌细胞向无功能去分化细胞的表型改变在约50年前就已在血管损伤中被发现，但转化细胞的表型和遗传身份仍不清楚。我们已经发现，肥大小肠中的转化细胞是肌成纤维细胞样细胞，其弱表达血小板衍生生长因子受体α和β（PDGFR α/β）（称为“PDGFR低β高细胞”）。我们从结肠和空肠中分离了分化的PDGFRahigh细胞和SMCs，并获得了基因组规模的基因表达谱。通过分析转录组，我们确定了SMC特异性转录因子，包括SRF和MYOCD。此外，我们发现SMC的表型受SRF和表观遗传调节因子DNA甲基转移酶1和3a（DNMT 1/3a）的调节。本项目旨在揭示一个细胞和遗传机制，以了解SMCs是如何在肠道SM肥大的发展过程中去分化，以及如何去分化的细胞在恢复过程中再分化成SMCs。此外，该项目旨在分离去分化细胞，建立培养细胞系，并使用SMC特异性转录因子和表观遗传调节因子DNMT 1/3a从细胞系中遗传工程化SMC。为了实现这些目标，我们与Seungil Ro博士（博士，PI），Moon young Lee博士组成了一个合作的多机构研究团队（MD/PhD，研究肥大条件下SMC的可塑性），待聘博士后（博士，研究PDGRFalow细胞的SMC工程），道格拉斯Redelman（PhD，细胞分离），Gabsang Lee（PhD/DVM，细胞系建立），Terence Smith（博士，钙成像），拉伦贝克尔（医学博士/博士，组织供应和咨询），肯特萨斯（医学博士，组织供应），约瑟夫米亚诺和塞隆桑德斯（博士，研究工具供应和项目咨询）。这些研究将显著推进我们对SMC在转录组水平上的表型转变的理解，并且可以提供用于临床的人SMC系，以修复在GI SM疾病中发现的受损或无功能的SMC。