Assessment of a potential application of endogenous stem cells to treat congenital disorders

评估内源干细胞治疗先天性疾病的潜在应用

• 批准号: 22K20740
• 负责人: Sunardi Mukhamad
• 金额: $ 1.83万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Research Activity Start-up
• 财政年份: 2022
• 资助国家: 日本
• 起止时间: 2022-08-31 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-22K20740/
• 关键词: tissue regeneration; HSCR; enteric nervous system; stem cell

Congenital diseases often result in incomplete organogenesis due to impaired stem cells function caused by toxic effects of gene mutations. Although surgical interventions are the primary treatment for these organ defects, their effectiveness remains unsatisfactory. However, advancements in genome editing technology now enable the in vivo correction of gene mutations in stem cells. To assess the potential of diseased stem cells to restore their ability to form autonomous tissues through gene repair, we successfully developed a mouse model of Hirschsprung disease (HSCR), serving as a model for congenital stem cell disorders. This model facilitates the removal of the disease-causing mutation through Cre-loxP-mediated gene recombination. The heterozygous mouse exhibited enteric nervous system defects resembling HSCR pathology. By crossing this mouse with Phox2b-Cre mice, we confirmed that deleting RET S811F in vagal neural crest-derived ENS stem cells evaded the enteric nerve defect phenotype. Moreover, crossing the HSCR mouse with Dhh-Cre mice eliminated the dominant negative mutation in Schwann cell precursors, which crucial for postnatal intestinal neurogenesis. We observed an increase in mCherry-positive intestinal neurons in regions lacking proper nerve function, suggesting compensation by cells where dominant-negative mutations were removed. The analysis of removing the disease-causing mutation in vagal neural crest-derived cells at different developmental time periods (Embryonic day 10.5. E11.5, and E12.5) revealed no notable differences among the observed time periods.

先天性疾病往往导致不完整的器官，由于受损的干细胞功能所造成的基因突变的毒性作用。虽然手术干预是这些器官缺陷的主要治疗方法，但其有效性仍不令人满意。然而，基因组编辑技术的进步现在能够在体内校正干细胞中的基因突变。为了评估患病干细胞通过基因修复恢复其形成自主组织的能力的潜力，我们成功地开发了先天性巨结肠病（HSCR）的小鼠模型，作为先天性干细胞疾病的模型。该模型有利于通过Cre-loxP介导的基因重组去除致病突变。杂合子小鼠表现出类似HSCR病理学的肠神经系统缺陷。通过将这只小鼠与Phox2b-Cre小鼠杂交，我们证实了在迷走神经嵴来源的ENS干细胞中删除RET S811F可以避免肠神经缺陷表型。此外，HSCR小鼠与Dhh-Cre小鼠杂交消除了雪旺细胞前体中的显性负突变，这对出生后的肠道神经发生至关重要。我们观察到在缺乏适当神经功能的区域中mCherry阳性肠神经元的增加，这表明显性阴性突变被去除的细胞的补偿。不同发育时期（胚胎10.5天）迷走神经嵴源性细胞致病突变的去除分析。E11.5和E12.5）显示在观察的时间段之间无显著差异。

项目成果

Potential application of endogenous stem cells for the treatment of congenital diseases

内源干细胞治疗先天性疾病的潜在应用

• 发表时间: 2023
• 作者: Peng Y;Tsuno Y;Matsui A;Hiraoka Y;Tanaka K;Horike S;Daikoku T;Mieda M;Mukhamad Sunardi
• 通讯作者: Mukhamad Sunardi