Uncovering the therapeutic potential of adipose tissue derived neural stem cells for Hirschsprung's disease.

揭示脂肪组织来源的神经干细胞治疗先天性巨结肠的潜力。

基本信息

批准号：
10580052
负责人：
ALLAN M GOLDSTEIN
金额：
$ 20.62万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10580052
关键词：
Address Adipose tissue Affect Blood flow Cell Separation Cells Central Nervous System Clinical Clinical Trials Coculture Techniques Colon Colonic Aganglionosis Colorectal Compensation Complex Congenital Megacolon Congenital Neuropathy Constipation Cues Data Deposition Disease Dose Engraftment Enteral Enteric Nervous System Enterocolitis Environment Excision Fatty acid glycerol esters Fecal Incontinence Fluorescence-Activated Cell Sorting Ganglia Gastrointestinal tract structure Goals Harvest Health Human In Vitro Intestinal Secretions Intestines Investigation Knowledge Laboratory culture Life Liquid substance Methods Microsurgery Morbidity - disease rate Movement Mus Nerve Fibers Nerve Tissue Nervous System Neuroglia Neurons Neuropathy Operative Surgical Procedures Patients Physiological Pluripotent Stem Cells Population Postoperative Complications Pre-Clinical Model Procedures Process Property Protocols documentation Rectum Regenerative Medicine Reporting Research Resolution Risk Schwann Cells Scientific Advances and Accomplishments Signal Transduction Smooth Muscle Source Suction Lipectomy System Therapeutic Tissues Transplantation Vasomotor adipose derived stem cell cell motility clinical application experimental study human stem cells human tissue implantation in vivo innovation interest migration minimally invasive mouse model nerve stem cell nervous system disorder neural neurogenesis novel patient safety postnatal postnatal human preservation prevent rectal response single nucleus RNA-sequencing stem cell biomarkers stem cell migration stem cell population stem cell therapy stem cells subcutaneous treatment strategy tumorigenic

项目摘要

PROJECT SUMMARY The overarching objective of our research is to develop a stem cell therapy from subcutaneous fat tissue (SAT) to replace the congenitally absent enteric nervous system (ENS) in Hirschsprung disease (HSCR). Surgical resection of the affected colon is currently the only viable treatment for HSCR. This is a necessary life-saving procedure; albeit, more than 50% of patients still suffer from postoperative complications including constipation, fecal incontinence, and enterocolitis. To overcome these morbidities, investigations into treatments that can preserve the rectum and its functions are warranted. Replacement of the absent ENS via stem cell therapies is touted as the most promising treatment strategy to achieve this goal. Our group has demonstrated the feasibility of stem cell treatments by harvesting neural stem cells (NSCs) from the gut which engraft, migrate and differentiate into neuronal networks when transplanted into mice with HSCR. For clinical application this would require surgical resection of a piece of intestine. To prevent unnecessary resection surgery, other sources of NSCs are of interest. Human fat (adipose) tissue contains a reservoir of stem cells that are readily accessible. These cells have been examined in over 270 clinical trials for numerous diseases that support favourable patient safety profiles. In our preliminary data we have also identified that nerve fiber bundles from murine fat deposits – subcutaneous adipose tissue (SAT) - harbor an endogenous source of NSCs that are unexplored for the treatment of neuropathies. We predict that the SAT could provide a useful source of NSCs to treat colonic aganglionosis in HSCR; however, it remains undetermined if SAT-NSCs can undergo neurogenesis in the aganglionic (absent ENS) environment of the gut and there are currently no methods to purify and expand human SAT-NSCs. In the first aim of this study, we will determine if purified SAT-NSCs from mice are capable of neurogenesis in aganglionic intestine. The ganglionated ENS is supplemented postnatally by NSCs that migrate into the gut from extrinsic nerve fiber bundles and differentiate into enteric neurons in response to environmental cues from the gut. We will address whether SAT-NSCs can also undergo enteric neurogenesis when provided signalling cues from the ganglionic and aganglionic gut in in vitro coculture systems and via microsurgical SAT-NSC implantation in vivo. To determine how to isolate and expand human SAT-NSCs we will address the paucity of knowledge on the origin of these cells. To accomplish this, cells isolated from human SAT nerve fiber bundles will be unbiasedly characterised by single nuclei RNA-Seq before and after stem cell culture procedures. Cells expressing NSC markers will be purified by fluorescence activated cell sorting and their differentiation potential will be assessed in in vitro culture and in ex vivo transplants to the smooth muscle of the gut. The results of these studies will establish procedures to isolate SAT-NSCs and assess their potential to treat the congenital neuropathy in HSCR.

项目摘要我们研究的总体目的是从皮下脂肪组织开发干细胞疗法（SAT）替代了Hirschsprung病（HSCR）中先天缺失的肠神经系统（ENS）。当前，受影响的结肠的手术切除是HSCR的唯一可行治疗方法。这是必要的挽救生命的程序；尽管有超过50％的患者仍患有术后并发症，包括便秘，粪便失禁和小肠结肠炎。为了克服这些病毒，调查可以保留可以保留直肠及其功能的治疗方法。替换缺席的ENS 干细胞疗法被吹捧为实现此目标的最有前途的治疗策略。我们的小组有通过从肠道中收集神经干细胞（NSC）来证明干细胞处理的可行性当用HSCR移植到小鼠中时，植入，迁移并分化为神经元网络。用于临床应用这将需要手术切除一块肠。防止不必要的切除手术，其他NSC的来源令人感兴趣。人脂肪（脂肪）组织含有干细胞的储层很容易访问。这些细胞已在270多种临床试验中检查了许多疾病支持有利的患者安全概况。在我们的初步数据中，我们还确定了神经纤维来自鼠脂肪沉积的捆绑包 - 皮下脂肪组织（SAT） - 内源性的来源对神经病的治疗意外的NSC。我们预测SAT可以提供有用的 NSC治疗HSCR中结肠诱发的NSC源；但是，如果SAT-NSC可以在肠道（缺乏ENS）环境中经历神经发生，目前没有净化和扩展人类SAT-NSC的方法。在这项研究的第一个目的中，我们将确定是否纯化来自小鼠的SAT-NSC能够在Aganglionic肠道中神经发生。神经节的ens是在产后补充NSC，这些NSC从外部神经纤维束中迁移到肠道中，响应于肠道的环境线索，分为肠神经元。我们将解决是否当提供神经节和神经节的信号线索时，SAT-NSC也可以接受肠神经发生 Aganglionic肠道体外共培养系统，并通过体内微表外sat-nsc植入。到确定如何隔离和扩展人类SAT-NSC，我们将解决有关知识的缺乏这些细胞的起源。为此，从人类SAT Never Fiber束中孤立的细胞将是公正的在干细胞培养程序之前和之后，以单核RNA-seq为特征。表达NSC的细胞标记将通过荧光激活的细胞分选纯化，其分化电位将是在体外培养和离体移植中评估肠道的平滑肌。这些结果研究将建立隔离SAT-NSC的程序并评估其治疗先天性的潜力 HSCR中的神经病。