Background/Aims Interstitial cells of Cajal (ICC) are specialized gastrointestinal (GI) pacemaker cells required for normal GI motility. Dysfunctions in ICC have been reported in patients with GI motility disorders, such as gastroparesis, who exhibit debilitating symptoms and greatly reduced quality of life. While the proteins, calcium-activated chloride channel anoctamin-1 (ANO1) and the receptor tyrosine kinase (KIT), are known to be expressed by human ICC, relatively little is known about the broad molecular circuitry underpinning human ICC functions. The present study therefore investigates the transcriptome and proteome of ANO1-expressing, KITlow/CD45-/CD11B- ICC obtained from primary human gastric tissue. Methods Excess human gastric tissue resections were obtained from sleeve gastrectomy patients. ICC were purified using fluorescence-activated cell sorting (FACSorting). Then, ICC were characterized by using immunofluorescence, real-time polymerase chain reaction, RNA-sequencing and mass spectrometry. Results Compared to unsorted cells, real-time polymerase chain reaction showed the KITlow/CD45-/CD11B- ICC had: a 9-fold (P < 0.05) increase in ANO1 expression; unchanged KIT expression; and reduced expression for genes associated with hematopoietic cells (CD68, > 10-fold, P < 0.001) and smooth muscle cells (DES, > 4-fold, P < 0.05). RNA-sequencing and gene ontology analyses of the KITlow/CD45-/CD11B- cells revealed a transcriptional profile consistent with ICC function. Similarly, mass spectrometry analyses of the KITlow/CD45-/CD11B- cells presented a proteomic profile consistent with ICC activities. STRING-based protein interaction analyses using the RNA-sequencing and proteomic datasets predicted protein networks consistent with ICC-associated pacemaker activity and ion transport. Conclusion These new and complementary datasets provide a valuable molecular framework for further understanding how ICC pacemaker activity regulates smooth muscle contraction in both normal GI tissue and GI motility disorders.

中文翻译：

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Cajal 原代人胃间质细胞的转录组和蛋白质组分析预测起搏器网络。

背景/目标 Cajal 间质细胞 (ICC) 是正常 GI 运动所需的特殊胃肠道 (GI) 起搏细胞。据报道，患有胃轻瘫等胃肠道运动障碍的患者会出现 ICC 功能障碍，这些患者表现出衰弱症状并大大降低生活质量。虽然已知蛋白质、钙激活氯离子通道 anoctamin-1 (ANO1) 和受体酪氨酸激酶 (KIT) 由人类 ICC 表达，但人们对支持人类 ICC 功能的广泛分子回路知之甚少。因此，本研究调查了从原代人胃组织中获得的表达 ANO1 的 KITlow/CD45-/CD11B-ICC 的转录组和蛋白质组。方法从袖状胃切除术患者中获得多余的人胃组织切除物。使用荧光激活细胞分选 (FACSorting) 纯化 ICC。然后，通过使用免疫荧光、实时聚合酶链反应、RNA 测序和质谱法对 ICC 进行表征。结果 与未分选的细胞相比，实时聚合酶链反应显示 KITlow/CD45-/CD11B- ICC 具有： ANO1 表达增加 9 倍 (P < 0.05)；不变的 KIT 表达；与造血细胞（CD68，> 10 倍，P < 0.001）和平滑肌细胞（DES，> 4 倍，P < 0.05）相关的基因表达降低。KITlow/CD45-/CD11B- 细胞的 RNA 测序和基因本体论分析揭示了与 ICC 功能一致的转录谱。同样，KITlow/CD45-/CD11B- 细胞的质谱分析呈现出与 ICC 活动一致的蛋白质组学特征。使用 RNA 测序和蛋白质组数据集的基于字符串的蛋白质相互作用分析预测与 ICC 相关起搏器活动和离子传输一致的蛋白质网络。结论 这些新的互补数据集为进一步了解 ICC 起搏器活动如何调节正常 GI 组织和 GI 运动障碍中的平滑肌收缩提供了有价值的分子框架。