Genes, environment & neural stem cell transplantation in the gut

基因、环境

• 批准号: 8926953
• 负责人: PANKAJ J PASRICHA
• 金额: $ 36.45万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8926953
• 关键词: Achalasia; Allogenic; Biological Factors; Cells; Clinical; Collagen; Collagen Type IV; Colonic Inertia; Congenital Megacolon; Disease; Dysplasia; Enteral; Enteric Nervous System; Environment; Ganglia; Gastrointestinal Motility; Gastroparesis; Genes; Health; In Vitro; Integrins; Methods; Muscle; Myenteric Plexus; Natural regeneration; Nerve; Nerve Regeneration; Neuroglia; Neurons; Neuropathy; PTEN gene; PTK2 gene; Pathway interactions; Preparation; Research Personnel; Role; Societies; Stem cell transplant; Transplantation; burden of illness; cell preparation; functional loss; human disease; in vivo; nerve stem cell; neural precursor cell; neurogenesis; novel therapeutic intervention

DESCRIPTION (provided by applicant): Loss of functional enteric neurons results in clinical disorders such as achalasia, gastroparesis, neuropathic forms of pseudoobstruction, enteric neuronal dysplasia, colonic inertia and Hirschsprung's disease. Despite the lack of significant neurogenesis in vivo in these conditions, there is growing evidence to suggest the existence of multipotent neural precursor cells (NPCs) in abundant numbers in the gut. Several investigators including ourselves have shown that these cells, which have some phenotypical markers typical of glia, are capable of robust neurogenesis in vitro. We have now shown that enteric ganglia in longitudinal muscle myenteric plexus (LMMP) preparations display neurogenesis and this is even more pronounced in completely dissociated NPC preparations in culture. We have identified enteric neural precursors in vivo and in vitro and our preliminary studies implicate phosphatase and tensin homolog (PTEN) in these cells as the major "brake" on neurogenesis in vivo. Further collagen IV when added to LMMP preparations can have a profound inhibitory effect on neurogenesis. Thus it is logical to hypothesize that the discrepancy between the in vivo and ex vivo situation can be explained by factors in the microenvironment that regulate PTEN. In this proposal we therefore intend to examine the effects of collagen IV and downstream pathways including integrin, FAK and RhoA/ROCK which may drive PTEN activity, checking enteric neurogenesis in vivo, via the following specific aims: Specific Aim 1: To determine the interactions between extrinsic (collagen) and intrinsic (PTEN) brakes on neurogenesis in health and disease Specific Aim 2: To examine the effects of countering intrinsic and extrinsic brakes on enteric neurogenesis in vivo in health and disease Specific Aim 3: To examine the effects of manipulating selected key components of these pathways on survival and function of allogeneic enteric NSC (ENSC) transplants Our long-term objective is to develop neuronal transplantation as a treatment for human diseases of the enteric nervous system as well as stimulate neural regeneration in these disorders. By establishing a role for PTEN and related pathways the proposed studies will provide the critical groundwork for novel therapeutic interventions.

描述（由申请人提供）：功能性肠神经元的丧失会导致临床疾病，例如腹部，胃轻瘫，神经植物结构的神经性形式，肠神经元不典型增生，结肠惯性和赫斯普伦病。尽管在这些条件下体内缺乏明显的神经发生，但越来越多的证据表明肠道中有大量的多能神经前体细胞（NPC）存在。包括我们自己在内的几个研究人员表明，这些具有典型的神经胶质表型标记的细胞能够在体外具有牢固的神经发生。现在，我们已经表明，纵向肌肉肌丛（LMMP）制剂中的肠神经节表现出神经发生，这在培养中完全分离的NPC制剂中更为明显。我们已经在体内和体外鉴定出肠神经前体，我们的初步研究暗示这些细胞中的磷酸酶和Tensin同源物（PTEN）是体内神经发生的主要“制动”。将胶原蛋白IV添加到LMMP制剂中时可能对神经发生产生深远的抑制作用。因此，假设体内和体内情况之间的差异可以通过调节PTEN的微环境中的因素来解释是合乎逻辑的。因此，在这项提案中，我们打算检查胶原蛋白IV和下游途径的影响，包括整联蛋白，FAK和RhoA/Rock，可能会驱动PTEN活动，检查体内的肠神经发生，通过以下特定目的：特定目的：特定目的1：确定缺血（胶原蛋白）和内在（PTEN）及其及其及其及其及其及其及其及其及其及其及其及其及其及其及其及其及其及其神经疾病的影响的相互作用，以实现对健康的影响，以实现对健康的影响，以实现健康的影响，以实现健康的影响，以实现健康的影响，以实现健康的影响，以实现健康的影响，以构成健康的脑部及其对健康的影响。在健康和疾病特定目的中对体内肠神经发生的制动3：检查操纵这些途径所选的关键组成部分对同种异体肠NSC（ENSC）的生存和功能的效果，我们的长期目标是发展神经元移植作为治疗人类疾病的神经元移植，并刺激这些企业的神经疾病，并刺激这些破坏性的神经疾病。通过确立PTEN和相关途径的作用，拟议的研究将为新的治疗干预措施提供关键的基础。