贲门失弛缓症是一种罕见病，病因不明，患者生活质量极差。免疫炎症引起的食管壁内抑制性肠神经丛损伤可能是贲门失弛缓症的发病基础。申请人前期：（1）通过单细胞转录组测序和多重免疫测定等手段发现，Th17细胞可能是贲门失弛缓症发病中的关键免疫亚群；（2）Th17细胞可引起共培养的神经元损伤，这一作用与外泌体分泌密切相关；（3）Th17细胞培养上清液外泌体中miR-101/199明显升高；下调miR-101/199表达后，可逆转其外泌体对共培养神经元的损伤。基于此，申请人提出Th17细胞通过外泌体miR-101/199介导抑制性肠神经元损伤而致病的假说。本研究拟:（1）验证Th17细胞对抑制性肠神经元损伤从而导致贲门失弛缓症的表型；（2）在细胞、动物和临床层面，探索Th17细胞通过外泌体miR-101/199引起抑制性肠神经元损伤及其具体机制。项目顺利进行将为揭示此病的病因提供新的依据。

Achalasia is a rare disease with extremely poor quality of life. Although with unknown etiology, immune-mediated inhibitory neuron degeneration has been proposed as one of the main causes. We previously found that Th17 cells may be the key subset in the pathogenesis of achalasia by single-cell RNA sequencing (scRNA-seq) and flow cytometry. SK-N-SH neurons were impaired while co-cultured with Th17 cells in vitro. Th17 cells derived exosomal miR-101/199 were highly expressed and could impair co-cultured neurons, which could be reversed by down-regulation of miR-101/199 of Th17 cells. Therefore, we propose that Th17 cells could mediate inhibitory enteric neuron degeneration by the exosomal miR-101/199. This study intends to: (1) verify Th17 cells could impair inhibitory enteric neurons and cause achalasia. (2) explore the mechanism of inhibitory enteric neuron damage caused by Th17 cells derived exosomal miR-101/199 using in vitro, animal model, and clinical samples. If successful, this study may provide new findings on the pathogenesis of achalasia.