Development of Bioactive Chemical Probes for Calcium-activated Chloride Channel

描述(申请人提供)：离子通道是膜蛋白，存在于可兴奋细胞和不可兴奋细胞中。它们选择性地引导离子穿过细胞膜，在细胞生理中发挥关键作用，包括电和细胞信号、离子动态平衡和激素分泌。这项建议的目的是为编码钙激活氯离子通道(CACC)的TMEM16A寻找有效和特异的小分子化学探针。CACC的异常被认为是囊性纤维化、哮喘、慢性支气管炎和高血压等疾病的原因。它的作用涉及许多生理过程，包括心脏和神经兴奋、感觉转导、跨上皮分泌、平滑肌收缩和受精。与首次克隆TMEM16A基因的莉莉·简博士的实验室合作，我们开发了几个细胞系和一种基于荧光的碘替代通量分析。用于高通量筛选的协议正在针对CACC目标TMEM16A进行测试。这项提案概述了一项计划，即使用表达TMEM16A的细胞系和遗传编码的Cl-/I-生物传感器进行&gt；300,000-化合物筛选，以寻找有效的CACC抑制/激活探针。然后，活性化合物将通过自动膜片钳记录进行评估。与加州大学旧金山分校的莉莉·简博士合作，我们未来的计划包括分别在神经元和上皮细胞等本地制剂中测试分离的化合物。随着药理学和药物化学的进一步努力，这些活性化合物可能被用作开发治疗CACC相关疾病的药物的工具或先导化合物。 与公共卫生相关：最近发现的钙激活氯离子通道(CACC)为识别小分子探针铺平了一条新的途径。CACC活性异常被认为是几种主要疾病的原因，如囊性纤维化、哮喘、慢性支气管炎和高血压。它的作用涉及许多生理过程，包括心脏和神经兴奋、感觉转导、跨上皮分泌、平滑肌收缩和受精。因此，特定的调制化合物对帮助治疗开发是有用的。

DESCRIPTION (provided by applicant): Ion channels are membrane proteins, found in both excitable cells and non-excitable cells. They selectively conduct ions across cellular membranes and play a critical role in cellular physiology, including electrical and cellular signaling, ion homeostasis, and hormone secretion. The objective of this proposal is to find potent and specific small molecule chemical probes for TMEM16A that encodes the calcium-activated chloride channel (CaCC). Abnormality of CaCC is thought to be causal to diseases, such as cystic fibrosis, asthma, chronic bronchitis, and hypertension. Its role is implicated in numerous physiological processes including cardiac and neuronal excitation, sensory transduction, trans-epithelial secretion, smooth muscle contraction, and fertilization. In collaboration with Dr. Lily Jan's laboratory, which first cloned the TMEM16A gene, we have developed several cell lines and a fluorescence-based iodide surrogate flux assay. The protocol for high-throughput screening is being tested for the CaCC target TMEM16A. This proposal outlines a plan to conduct a >300,000-compound screen using the TMEM16A expressing cell line with a genetically encoded Cl-/I- biosensor to search potent inhibitor/activator probes for CaCC. The active compounds will then be evaluated by automated patch-clamp recording. In collaboration with Dr. Lily Jan at University of California at San Francisco, our future plan includes testing of the isolated compounds in native preparations, such as neurons and epithelium, respectively. These active compounds, with further efforts in pharmacology and medicinal chemistry, may be exploited either as tools or as lead compounds for developing therapeutic remedies to the CaCC-associated diseases. PUBLIC HEALTH RELEVANCE: The recent discovery of the calcium-activated chloride channel (CaCC) has paved a new path to identify small molecule probes. Abnormality of CaCC activity is thought to be causal to several major diseases, such as cystic fibrosis, asthma, chronic bronchitis, and hypertension. Its role is implicated in numerous physiological processes including cardiac and neuronal excitation, sensory transduction, trans-epithelial secretion, smooth muscle contraction, and fertilization. Therefore, specific modulation compounds are useful to aid therapeutic development.