Calcium activated chloride channel modulation of myometrial excitability

 描述（由申请人提供）：早产是一个巨大的，日益增长的全球卫生保健挑战。在美国，每年发生> 520，000例早产，其中许多与相当高的发病率相关。尽管研究已经深入了解了导致早产（PTL）的机制，但PTL的治疗手段仍然有限且无效。然而，最近对钙激活氯离子通道的Anoctamin（ANO）家族的鉴定彻底改变了我们对这些氯离子通道如何在调节其他平滑肌组织的膜电位、起搏和收缩性中发挥不可或缺的作用的理解。在目前的提案中，我们建立了ANO1在人类子宫平滑肌（USM）的收缩性和起搏频率中的功能重要性，确定了使用ANO1作为体外减弱人类USM收缩的治疗靶点的可行性，并探索了负责这些有益作用的潜在分子机制。特别是，研究将跨越整个组织功能器官浴实验 对从妊娠晚期剖腹产患者采集的新鲜人子宫肌条进行了基于细胞的机制研究，重点关注这些通道在驱动USM细胞膜电位、钙处理和非特异性肌动蛋白动力学中的作用。我们将采用广泛的技术，包括共聚焦显微镜，电生理学，体内和体外基因编辑，以及药理学拮抗剂策略，以确定该通道家族在人类子宫收缩中的作用。这项工作的目的不仅是建立这个通道在正常子宫平滑肌生理学中的重要性，而且还确定它是否可以作为治疗过早收缩的新靶点。因此，高度翻译的可能性 存在早产的治疗方法。

 DESCRIPTION (provided by applicant): Preterm birth is an enormous, growing global health care challenge. In the United States, >520,000 preterm births occur each year, many of which are associated with substantial morbidity. Although research has made insights into the mechanisms contributing to pre-term labor (PTL), the therapeutic armamentarium for PTL remains limited and inefficacious. However, the recent identification of the anoctamin (ANO) family of calcium-activated chloride channels has revolutionized our understanding of how these chloride channels play integral roles in modulating membrane potential, pacing, and contractility in other smooth muscle tissues. In this current proposal we establish ANO1's functional importance in the contractility and pacing frequency of human uterine smooth muscle (USM), determine the feasibility of using ANO1 as a therapeutic target to attenuate human USM contractions in vitro, and explore the underlying molecular mechanisms responsible for these beneficial effects. In particular, studies will span whole tissue functional organ bath experiments on fresh human myometrium strips harvested from late gestation c-section patients to mechanistic cell based studies focusing on the role of these channels on driving USM cell membrane potential, in calcium handling and filamentous-actin dynamics. We will employ a wide range of techniques including confocal microscopy, electrophysiology, in vivo and in vitro gene editing, and pharmacological antagonist strategies to establish the role of this channel family in human uterine contractility. The work is purposed to not only establish the importance of this channel in normal uterine smooth muscle physiology, but to also ascertain if it may serve as a new target to treat pre-mature contractions. As such, the potential for a highly translational therapy for pre- term labor exists.