Developing a human pluripotent stem cell-based strategy for treating Hirschsprung disease

开发基于人类多能干细胞的策略来治疗先天性巨结肠症

• 批准号: MR/V002163/1
• 负责人: Anestis Tsakiridis
• 金额: $ 129.6万
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FV002163%2F1
• 关键词: Developing; human; pluripotent; stem; cell

Hirschsprung disease is a life-threatening intestinal disorder caused by an absence of intrinsic nerve cells (aganglionosis) in the most distal gastrointestinal tract. It occurs in approximately 1 in 5000 live births, making it one of the most common congenital diseases affecting the gut. Given that intrinsic gut nerve cells (enteric neurons) mediate the contractions necessary for normal gut function, their absence in Hirschsprung patients causes severe constipation or intestinal obstruction. The only treatment available is surgical removal of the affected part of the bowel combined with a 'pull through' procedure, which entails connecting the healthy part of the gut to the anus. However, the surgery necessitates retention of part of the abnormal gut including the anal sphincter, which is likely to account in part for the long-term, often life-long, gastrointestinal problems and poor quality of life suffered by the majority of patients with Hirschsprung disease. Surgery, readmissions and outpatients hospital appointments required for management of this condition present a significant burden for the healthcare system. Recent advances in the understanding of development of the gut's intrinsic (or enteric) nervous system and pathogenesis of the disease, as well as considerable progress in regenerative medicine, have highlighted potential for alternative treatments, such as cell replacement therapy.During normal embryonic development, the enteric nervous system is derived from a transient population of cells termed neural crest cells that migrate from the neural tube to innervate the gastrointestinal tract. In Hirschsprung disease these cells fail to colonize the distal gut leaving a variable segment of aganglionosis. Experiments in animal models have demonstrated that transplantation of enteric nervous system progenitor/stem cells (derivatives of the original neural crest cells and harvested from mouse gut explants or mouse embryonic stem cells) have the ability to differentiate into enteric neurons and colonize when transplanted into aganglionic gut explants. Similar neuronal progenitor cells obtained from human postnatal gut explants have been shown to effectively colonize and differentiate within aganglionic gut explants of Hirschsprung patients. However, the harvesting of such human nerve progenitor cells from gut is difficult and becomes more so with increasing post-natal age. Therefore, although such preclinical testing has demonstrated that cell therapy should be a viable option for curing Hirschsprung disease, the availability of human enteric neurons from post-natal gut remains a bottleneck for development of cell-based therapies.An attractive alternative source is offered by pluripotent stem cells, where there is significant potential of generating appropriate cells for therapy both in terms of numbers and tailoring cell properties for the task. We have recently developed efficient protocols to generate human neural crest cells and enteric nervous system progenitors from human pluripotent stem cells. In this project, we will test the ability of these progenitors to correct for the lack of enteric neurons in animal and patient-derived models of Hirschsprung disease following transplantation. We will also optimise the current methods of generating and purifying human enteric nervous system progenitors from pluripotent stem cellsand will develop a new improved animal model of Hirschsprung disease. In this way, we will establish the pre-clinical basis for a regenerative medicine approach for treating Hirschsprung disease by transplantation of cells derived from human pluripotent stem cells to re-innervate the affected part of the gut. In the first instance our goal will be to improve the outcome of current treatments by innervation of the short section remaining after routine surgery.

先天性巨结肠症是一种危及生命的肠道疾病，由最远端胃肠道缺乏内在神经细胞（神经节细胞增多症）引起。它发生在大约五千分之一的活产中，使其成为影响肠道的最常见先天性疾病之一。鉴于内在的肠道神经细胞（肠神经元）介导正常肠道功能所需的收缩，先天性巨结肠患者缺乏它们会导致严重的便秘或肠梗阻。唯一可用的治疗方法是手术切除肠道受影响的部分，并结合“拉通”手术，将肠道的健康部分连接到肛门。然而，手术需要保留部分异常肠道，包括肛门括约肌，这可能是大多数先天性巨结肠患者遭受长期（通常是终生）胃肠道问题和生活质量差的部分原因。治疗这种疾病所需的手术、再入院和门诊预约给医疗保健系统带来了沉重的负担。对肠道内在（或肠）神经系统发育和疾病发病机制的理解的最新进展，以及再生医学的巨大进展，凸显了替代疗法（例如细胞替代疗法）的潜力。在正常胚胎发育过程中，肠神经系统源自称为神经嵴细胞的瞬时细胞群，这些细胞从神经管迁移以神经支配胃肠道。在先天性巨结肠症中，这些细胞无法定植于远端肠道，留下一段可变的无神经节细胞症。动物模型实验表明，肠神经系统祖细胞/干细胞（原始神经嵴细胞的衍生物，从小鼠肠道外植体或小鼠胚胎干细胞中收获）的移植具有分化为肠神经元的能力，并在移植到无神经节肠外植体中时定植。从人出生后肠道外植体中获得的类似神经元祖细胞已被证明可以在先天性巨结肠患者的无神经节肠道外植体中有效定植和分化。然而，从肠道中获取此类人类神经祖细胞很困难，并且随着出生后年龄的增加变得更加困难。因此，尽管此类临床前测试已证明细胞疗法应该是治疗先天性巨结肠症的可行选择，但来自出生后肠道的人类肠道神经元的可用性仍然是基于细胞的疗法开发的瓶颈。多能干细胞提供了一个有吸引力的替代来源，其在产生用于治疗的适当细胞的数量和定制细胞特性方面具有巨大的潜力。我们最近开发了有效的方案，从人类多能干细胞中产生人类神经嵴细胞和肠神经系统祖细胞。在这个项目中，我们将测试这些祖细胞在移植后纠正动物和患者来源的先天性巨结肠模型中肠神经元缺乏的能力。我们还将优化目前从多能干细胞中产生和纯化人类肠神经系统祖细胞的方法，并将开发一种新的改进的先天性巨结肠动物模型。通过这种方式，我们将为通过移植源自人类多能干细胞的细胞来重新刺激肠道受影响部分来治疗先天性巨结肠的再生医学方法奠定临床前基础。首先，我们的目标是通过常规手术后剩余的短节的神经支配来改善当前治疗的效果。

项目成果

Early anteroposterior regionalisation of human neural crest is shaped by a pro-mesodermal factor.

• DOI: 10.7554/elife.74263
• 发表时间: 2022-09-26
• 期刊: eLife
• 影响因子: 7.7
• 作者: Gogolou A;Souilhol C;Granata I;Wymeersch FJ;Manipur I;Wind M;Frith TJR;Guarini M;Bertero A;Bock C;Halbritter F;Takasato M;Guarracino MR;Tsakiridis A
• 通讯作者: Tsakiridis A

Early anteroposterior regionalisation of human neural crest is shaped by a pro-mesodermal factor

人类神经嵴的早期前后区域化是由前中胚层因子塑造的

• DOI: 10.1101/2021.09.24.461516
• 发表时间: 2021
• 作者: Gogolou A

A short history of pluripotent stem cells markers.

• DOI: 10.1016/j.stemcr.2023.11.012
• 发表时间: 2024-01-09
• 期刊: STEM CELL REPORTS
• 影响因子: 5.9
• 作者: Andrews, Peter W.;Gokhale, Paul J.
• 通讯作者: Gokhale, Paul J.

Notch signalling influences cell fate decisions and HOX gene induction in axial progenitors

Notch信号传导影响轴向祖细胞的细胞命运决定和HOX基因诱导

• DOI: 10.1101/2023.06.16.545269
• 发表时间: 2023
• 作者: Cooper F

Shaping axial identity during human pluripotent stem cell differentiation to neural crest cells.

• DOI: 10.1042/bst20211152
• 发表时间: 2022-02-28
• 期刊: Biochemical Society transactions
• 影响因子: 3.9