Autologous cell transplantation for the treatment of colorectal aganglionosis

自体细胞移植治疗结直肠无神经节细胞病

基本信息

批准号：
10308113
负责人：
Ryo Hotta
金额：
$ 7.75万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10308113
关键词：
Action Potentials Address Affect Age Area Autologous Autologous Transplantation Cell Separation Cell Survival Cell Therapy Cell Transplantation Cells Child Childhood Chronic Clinical Colon Colorectal Congenital Disorders Congenital Megacolon Constipation Data Development Diphtheria Toxin Disease Distal Drug Screening Electrophysiology (science)Embryo Enteral Enteric Nervous System Enterocolitis Environment Esophageal achalasia Excision Exhibits Failure Fecal Incontinence Functional Gastrointestinal Disorders Functional disorder Gastrointestinal Diseases Gastrointestinal tract structure Gastroparesis Goals Growth Factor Healthcare Human Immunosuppression In Vitro Injections Intestinal Pseudo-Obstruction Intestines Irritable Bowel Syndrome Laboratories Length Life Mediating Methods Modeling Morbidity - disease rate Mus Nerve Neural Crest Neuroglia Neuronal Differentiation Neuronal Injury Neurons Only Child Operative Surgical Procedures Proliferating Rodent Savings Stem cell transplant Synapses Testing Time Transgenic Mice Transplantation base cell motility clinical application clinically significant combinatorial cost diphtheria toxin receptor effective therapy enteric neuropathy experimental study ganglion cell gastrointestinal gastrointestinal function in vivo induced pluripotent stem cell innovation migration mouse model nerve stem cell novel novel strategies postnatal progenitor reduce symptoms single-cell RNA sequencing stem stem cells transcriptome transcriptomics transplantation therapy

项目摘要

PROJECT SUMMARY Hirschsprung disease is a potentially lethal congenital disorder characterized by the absence of enteric nervous system (ENS) along variable lengths of distal intestine due to the failure of neural crest-derived cells to colonize the entire intestine during development. The surgical treatment of Hirschsprung disease involves removing the aganglionic segment. While this is life- saving, significant complications commonly occur after surgery, including constipation, fecal incontinence, and enterocolitis. Transplantation of enteric neural crest-derived cells (ENCCs) offers a promising new approach to replacing missing or abnormal enteric neurons in Hirschsprung disease and other neurointestinal diseases. Recent studies, including from our laboratory, demonstrate that ENCCs can be isolated from the postnatal intestine, propagated in culture, and transplanted into the gut wall. However, major challenges remain: (1) the numbers of neurons generated have been limited, (2) their ability to integrate into neuroglial networks capable of restoring gut function has yet to be demonstrated, and (3) mice with Hirschsprung disease die at 4-6 weeks of age, limiting the time window available for analysis after cell transplantation. To overcome these challenges, we propose the following aims: (1) to incorporate an innovative approach to isolating and expanding postnatal gut-derived ENCCs that generates greater numbers of progenitor ENCCs than prior methods and (2) to transplant autologously-derived ENCCs into a novel non-lethal model of colorectal aganglionosis generated by local injection of human diphtheria toxin (DT) into transgenic mice whose neural crest-derived cells express DT receptor. To optimize ENCC expansion, a combinatorial drug screen approach will be used. ENCCs will be cultured with candidate molecules, together with known growth factors, to identify the optimal “cocktail” for progenitor cell expansion. Single cell RNAseq will be performed to characterize the transcriptome profile of cells prior to transplantation. Autologously-derived donor cells will be delivered into the experimentally- generated aganglionic segment of colon. Analyses will include quantitative determination of ENCC survival, proliferation, and neuronal differentiation, and functional characterization of neuronal activity. The results obtained will establish an optimized approach to ENCC isolation, expansion, and transplantation, and demonstrate the potential of cell-based therapy to restore GI function in Hirschsprung disease and other neurointestinal diseases.

项目摘要 Hirschsprung病是一种潜在的致命先天性疾病，其特征是没有由于失败神经rest衍生的细胞在发育过程中殖民整个肠道。手术赫希斯普伦疾病的治疗涉及去除僵化的节段。虽然这是生命 - 手术后通常会发生储蓄，明显的并发症，包括便秘，粪便尿失禁和小肠结肠炎。肠神经元衍生细胞（ENCC）的移植在替换缺失或异常的肠神经元的新方法中提供了承诺的新方法 Hirschsprung疾病和其他神经疾病。最近的研究，包括我们实验室，证明可以将ENCC与产后肠道隔离，在培养，并移植到肠壁上。但是，仍然存在主要挑战：（1）数字产生的神经元受到限制，（2）它们整合到神经网络中的能力能够恢复肠道功能尚未证明，并且（3）带有Hirschsprung的小鼠疾病在4-6周龄时死亡，限制了用于分析后的时间窗口移植。为了克服这些挑战，我们提出以下目的：（1）纳入了一种创新的方法，用于隔离和扩展产后肠道衍生的ENCC 比先前的方法生成更多的祖细胞ENCC，并且（2）移植自体衍生的ENCC成为一种新型的非致命性结肠直肠体病模型通过局部注射人二乳毒素毒素（DT）产生的转基因小鼠的神经波峰衍生的细胞表达DT接收器。为了优化ENCC扩展，一种组合药物将使用屏幕方法。 ENCC将与候选分子一起培养已知的生长因子，以确定用于祖细胞扩张的最佳“鸡尾酒”。单细胞将执行RNASEQ以表征细胞的转录组轮廓移植。自体源细胞将在实验中传递到结肠的产生的Aganglionic片段。分析将包括定量确定 ENCC存活，增殖和神经元分化以及功能表征神经元活性。获得的结果将建立一种优化的方法来隔离，扩展和移植，并证明基于细胞的治疗的潜力恢复 Hirschsprung疾病和其他神经疾病疾病中的GI功能。