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.

项目总结： 先天性或获得性肠道神经系统(ENS)中断会导致神经肠道疾病， 包括先天性巨结肠、食道失弛缓症、胃轻瘫、慢性假性肠梗阻、 还有其他人。功能性ENS的正常发育依赖于肠道之间的协调相互作用 神经沟来源的细胞及其微环境，包括作为肠道的细胞外基质(ECM) 神经沟来源的细胞沿着胚胎肠道间充质向尾端迁移。 确保适当的细胞迁移、增殖和分化为肠道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 细胞外基质结合蛋白已被证明能促进肠神经嵴来源的细胞迁移，而其他 (包括聚集素和硫酸软骨素蛋白多糖)已被证明抑制迁移，证明了一种 ECM在ENS形成中的关键作用。尽管对正常的ENS的发育有了更好的理解， 目前治疗神经性肠道疾病的方法缺乏，因为它们针对的是症状而不是潜在的 无ENS或异常ENS的病理学。我们的实验室和其他人正在研究肠道神经元的潜力 干细胞移植是恢复这些患者神经和肠道功能的一种新方法，但 移植细胞的植入、迁移和增殖有限阻碍了研究。 ENS的发展依赖于当地的环境，我们相信ENTERIC的成功发展 神经干细胞移植依赖于允许的局部ECM来支持移植细胞。我们 假设抑制性细胞外基质蛋白(集聚蛋白和硫酸软骨素蛋白多糖)过度表达 肠神经干细胞移植后，阻碍干细胞的迁移。在这笔赠款中，我们 因此，目的确定无神经节段结肠和正常结肠之间ECM基因表达的差异。 肠神经干细胞移植后ECM蛋白表达的特征，并利用我们的 了解正常ENS发育以改善肠神经干细胞后的微环境 通过击倒抑制细胞迁移的ECM蛋白进行移植，以增强细胞的迁移 移植的细胞。