Modifying the intestinal extracellular matrix to optimize enteric neuronal stem cell therapy for Hirschsprung disease

修饰肠细胞外基质以优化先天性巨结肠症的肠神经元干细胞疗法

• 批准号: 10731056
• 负责人: Jessica Mueller
• 金额: $ 8.31万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10731056
• 关键词: Agrin; Autologous Stem Cell Transplantation; Binding Proteins; Cell Migration Inhibition function; Cell Therapy; Cell Transplantation; Cells; Chondroitin Sulfate Proteoglycan; Chondroitinases; Chronic; Collagen; Collagen Type VI; Colon; Congenital Megacolon; Data; Development; Disease; Distal; Embryo; Engraftment; Ensure; Enteral; Enteric Nervous System; Environment; Esophageal achalasia; Extracellular Matrix; Extracellular Matrix Proteins; Fibronectins; Gastrointestinal Process; Gastroparesis; Gene Expression; Goals; Grant; Human; Integrins; Intestinal Pseudo-Obstruction; Intestines; Laminin; Lentivirus Vector; Mesenchyme; Modeling; Morbidity - disease rate; Motor; Mus; N-Cadherin; Neural Crest; Neuroglia; Neurons; Pathology; Patients; Pharmacotherapy; Process; Proliferating; Proteins; Research; Role; Sensory; Stem cell transplant; Symptoms; Tenascin; Transplantation; Vitronectin; blastomere structure; cell motility; embryo cell; improved; knock-down; migration; mouse model; nanoparticle; nerve stem cell; nerve supply; nervous system development; novel; novel strategies; overexpression; permissiveness; postnatal; protein expression; small hairpin RNA; stem cell migration; stem cell therapy; success; transcriptome sequencing; versican

PROJECT SUMMARY: Congenital or acquired disruption of the enteric nervous system (ENS) leads to neurointestinal diseases, including Hirschsprung disease, esophageal achalasia, gastroparesis, chronic intestinal pseudo-obstruction, and others. Normal development of a functional ENS relies on the coordinated interaction between enteric neural crest-derived cells and their microenvironment, including the extracellular matrix (ECM), as the enteric neural crest-derived cells migrate rostrocaudally along the embryonic gut mesenchyme.1 These interactions ensure proper cell migration, proliferation, and differentiation into the enteric neurons and glial cells that regulate numerous gastrointestinal processes, including motor and sensory function. Numerous ECM and ECM binding proteins have been shown to promote enteric neural crest-derived cell migration, while others (including agrin and chondroitin sulfate proteoglycans) have been shown to inhibit migration, demonstrating a critical role of the ECM in ENS formation. Despite an improving understanding of normal ENS development, current therapies for neurointestinal diseases are lacking, as they target symptoms rather than the underlying pathology of an absent or abnormal ENS. Our lab and others are investigating the potential of enteric neuronal stem cell transplantation as a novel approach to restoring innervation and gut function in these patients, but studies have been hampered by limited engraftment, migration, and proliferation of transplanted cells.1 Given the reliance of ENS development on the local environment, we believe that successful development of enteric neuronal stem cell transplantation relies on a permissive local ECM to support the transplanted cells. We hypothesize that inhibitory ECM proteins (agrin and chondroitin sulfate proteoglycans) are over-expressed following enteric neuronal stem cell transplantation, thwarting the migration of stem cells. In this grant we therefore aim to identify ECM gene expression differences between the aganglionic and normal colon, characterize ECM protein expression following enteric neuronal stem cell transplantation, and to leverage our understanding of normal ENS development to modify the microenvironment after enteric neuronal stem cell transplantation by knocking down ECM proteins that inhibit cell migration in order to enhance the migration of transplanted cells.

项目概要： 先天性或后天性肠神经系统 (ENS) 破坏会导致神经肠道疾病， 包括先天性巨结肠症、食管贲门失弛缓症、胃轻瘫、慢性假性肠梗阻、 和其他人。功能性 ENS 的正常发育依赖于肠溶之间的协调相互作用 神经嵴衍生细胞及其微环境，包括细胞外基质（ECM），作为肠 神经嵴衍生细胞沿着胚胎肠道间充质向尾部迁移。1 这些相互作用 确保细胞适当迁移、增殖和分化为肠神经元和神经胶质细胞 调节许多胃肠道过程，包括运动和感觉功能。众多 ECM 和 ECM 结合蛋白已被证明可以促进肠神经嵴衍生的细胞迁移，而其他蛋白 （包括集聚蛋白和硫酸软骨素蛋白多糖）已被证明可以抑制迁移，证明 ECM 在 ENS 形成中的关键作用。尽管人们对 ENS 正常发展的了解不断加深， 目前神经肠道疾病的治疗方法还很缺乏，因为它们针对的是症状而不是根本原因 ENS 缺失或异常的病理学。我们的实验室和其他人正在研究肠神经元的潜力 干细胞移植作为恢复这些患者神经支配和肠道功能的新方法，但是 研究因移植细胞的植入、迁移和增殖有限而受到阻碍。1 ENS的发展对当地环境的依赖，我们相信肠溶的成功发展 神经元干细胞移植依赖于允许的局部 ECM 来支持移植的细胞。我们 假设抑制性 ECM 蛋白（集聚蛋白和硫酸软骨素蛋白聚糖）过度表达 肠神经元干细胞移植后，阻碍干细胞的迁移。在这笔赠款中我们 因此，旨在确定无神经节结肠和正常结肠之间 ECM 基因表达的差异， 表征肠神经元干细胞移植后的 ECM 蛋白表达，并利用我们的 了解正常ENS发育以改变肠神经元干细胞后的微环境 移植通过敲低抑制细胞迁移的ECM蛋白来增强细胞的迁移 移植的细胞。