Piezo-1 & 2’s role in murine intestinal muscularis cells of the SIP syncytium

压电1

• 批准号: 10587233
• 负责人: James C Dunn
• 金额: $ 74.95万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10587233
• 关键词: Acute; Applications Grants; Biological Models; Biomechanics; Biomedical Engineering; Calcium; Calcium Channel; Cells; Cellular Structures; Clinical; Cues; Data; Disease; Enteral; Enterochromaffin Cells; Enteroendocrine Cell; Epithelium; Event; Failure; Frequencies; Gap Junctions; Genetic; Giant Cells; Goals; Growth; Hydrogels; Impairment; In Vitro; Interstitial Cell of Cajal; Intestinal Obstruction; Intestines; Ion Channel; Knock-out; Mechanics; Mediating; Methods; Modeling; Molecular; Morphology; Mus; Myopathy; Neuroglia; Neurons; Obstruction; Patients; Phase; Physiological; Piezo 1 ion channel; Pilot Projects; Platelet-Derived Growth Factor Receptor; Process; Property; Receptor Cell; Research; Role; Small Intestines; Smooth Muscle Myocytes; Stretching; System; Testing; Therapeutic; Thick; Tracer; Villus; adverse outcome; bioelectricity; cell motility; cell type; comparison control; cost; effective therapy; experimental study; genetic approach; in vitro Model; in vivo; innovation; insight; intestinal epithelium; mechanical force; mechanotransduction; motility disorder; nodal myocyte; novel; release of sequestered calcium ion into cytoplasm; response; scaffold; sensor; stem; tool; transmission process

This grant application aims to identify and characterize the role of Piezo in cells of the muscularis during baseline and stretched states. Bowel motility disorders and acute bouts of bowel obstruction are common clinical problems and can be severe, and we do not fully understand their mechanisms or have effective treatments. One crucial mechanism controlling bowel motility is mechanosensing, allowing each bowel segment to sense and respond to stretch. Various cell types within the intestinal muscularis control motility, sense stretch, and induce contractions. Among these cells are the components of the SIP syncytium comprised of smooth muscle cells (SMC), interstitial cells of Cajal (ICC), and PDGFRcells. These cells respond to stretch and receive inputs from enteric neurons (EN), glia, and enteroendocrine cells (EEC) that modulate SIP syncytium activity. However, the mechanism underlying the ability to sense and respond to stretch has not been thoroughly evaluated. Here we propose to investigate the role of Piezo in this process. Piezo is a relatively new calcium channel that functions as a mechanosensor in numerous cells, including those in the gut. Here, we will focus on its role in smooth muscle cells, interstitial cells of Cajal, and PDGFRcells of the small intestine. To perform these experiments, we have developed novel methods to maintain murine cells of the muscularis in an in vitro system. We will also use physiologically tunable hydrogels to examine the consequences of stretch in this setting. Finally, we will use various genetic approaches to explore the effects of mechanosensing in SIP cells of the gut. After this study, we anticipate that we will further elucidate the biomechanical and physiological consequences of disrupting a mechanosensitive channel that alters muscularis function and highlight myopathy's indirect effect on the adjacent epithelial layer to exacerbate the dysmotility further.

本拨款申请旨在确定和表征基线和拉伸状态下压电在肌层细胞中的作用。肠蠕动障碍和急性肠梗阻是常见的临床问题，并且可能很严重，我们尚未完全了解其机制或情况。控制肠道蠕动的一个关键机制是机械传感，使每个肠道节段都能感知拉伸并做出反应。 肠肌层内的各种细胞类型控制运动、感知拉伸并诱导收缩，其中包括由平滑肌细胞 (SMC)、Cajal 间质细胞 (ICC) 和 PDGFR 细胞组成的 SIP 合胞体。这些细胞对拉伸做出反应，并接收来自调节 SIP 合胞体活性的肠神经元 (EN)、神经胶质细胞和肠内分泌细胞 (EEC) 的输入。感知和响应拉伸的能力尚未得到彻底评估，压电是一种相对较新的钙通道，在许多细胞（包括肠道细胞）中发挥机械传感器的作用。在这里，我们将重点关注其在小肠平滑肌细胞、Cajal 间质细胞和 PDGFR细胞中的作用。 为了进行这些实验，我们开发了在体外系统中维持小鼠肌层细胞的新方法，我们还将使用生理可调水凝胶来检查拉伸在这种情况下的后果。机械感应对肠道 SIP 细胞的影响 在这项研究之后，我们预计我们将进一步阐明破坏改变肌层功能的机械感应通道的生物力学和生理学后果，并强调肌病的间接影响。对邻近上皮层的影响进一步加剧运动障碍。