LPA2 receptor-containing complexes in regulating secretory diarrhea

含 LPA2 受体的复合物调节分泌性腹泻

• 批准号: 9284460
• 负责人: Anjaparavanda P Naren
• 金额: $ 35.1万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-15 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9284460
• 关键词: Address; Adenylate Cyclase; Affect; Africa; Agonist; Americas; Apical; Applications Grants; Asia; Attenuated; Bacillus (bacterium); Biological Assay; Blood; Body Weight decreased; Cell membrane; Cells; Cessation of life; Chloride Channels; Chlorides; Cholera; Cholera Toxin; Complex; Country; Coupled; Cyclic AMP; Cystic Fibrosis Transmembrane Conductance Regulator; Diarrhea; Disease; Distal; Edg4 Protein; Enterotoxins; Epidemic; Epithelial Cells; Fluids and Secretions; Food; Food Contamination; Future; G-substrate; GTP-Binding Proteins; Generations; Haiti; Human; Inflammatory Bowel Diseases; Intestines; Latin America; Lysophosphatidic Acid Receptors; Macromolecular Complexes; Mediating; Metabolic; Methods; Microscopy; Mus; Pattern; Phosphorylation; Physiological; Play; Protein Isoforms; Proteomics; Reporting; Role; Scaffolding Protein; Small Interfering RNA; Stem cells; Symptoms; Testing; Therapeutic; Vibrio cholerae; World Health Organization; analog; assay development; base; clinically relevant; contaminated water; drug discovery; high throughput screening; ileum; intestinal crypt; lipid mediator; lysophosphatidic acid; multidisciplinary; public health relevance; small molecule; transcriptome; transcriptome sequencing

 DESCRIPTION (provided by applicant): The unifying hypothesis for this grant application is that LPA2 agonists have the potential for therapy of cholera and other types of enterotoxin-induced secretory diarrhea. Thus in this renewal application we propose to develop a physiologically relevant human intestinal stem cells (enteroids) to study CFTR-dependent fluid secretion and develop assays to demonstrate and identify LPA2 receptor specific small molecule agonists. LPA2 dependent inhibition of CFTR requires adenylate cyclase 6 (AC6) suggesting that AC6 is part of the CFTR-NHERF2-LPA2 complex. Two hypotheses will be tested: Specific Aim 1: To test the hypothesis that LPA2 agonists attenuate CTX-induced and CFTR- dependent fluid secretion in human intestinal stem cells (spheroids and enteroids), and mitigate diarrheal symptom Specific Aim 2: To test the hypothesis that adenylate cyclase 6 (AC6) exists in the macromolecular complex of CFTR-NHERF2-LPA2 in human intestinal epithelial cells and that AC6 plays an important role in modulating LPA2-dependent inhibition of CFTR channel function that can be therapeutically relevant in targeting LPA2 in controlling secretory diarrheas. The proposed study is highly significant because (a) it addresses a deadly disease; (b) it has clinical relevance and implications; (c) it is a multidisciplinary project coves basic biomedical studies, high throughput assay development, and sets a stage for future drug discovery.

 描述（由申请人提供）：本授权申请的统一假设是LPA2激动剂具有治疗霍乱和其他类型肠毒素诱导的分泌性腹泻的潜力。因此，在该更新申请中，我们提出开发生理学相关的人肠干细胞（肠样细胞）以研究CFTR依赖性液体分泌，并开发用于证明和鉴定LPA2受体特异性小分子激动剂的测定法。CFTR的LPA 2依赖性抑制需要腺苷酸环化酶6（AC6），这表明AC6是CFTR-NHERF2-LPA 2复合物的一部分。将检验两个假设：具体目标1：为了检验LPA2激动剂减弱人肠干细胞中CTX诱导的和CFTR依赖的液体分泌的假设，（球状体和肠状体），并减轻腹泻症状具体目标2：为了验证腺苷酸环化酶6（AC6）存在于人肠上皮细胞CFTR-NHERF 2-LPA 2大分子复合物中，并且AC6在调节LPA 2-NHERF 2中起重要作用的假设。CFTR通道功能的依赖性抑制，其在控制分泌型糖尿病中靶向LPA 2方面可以是治疗相关的。拟议的研究是非常重要的，因为（a）它解决了一个致命的疾病;（B）它具有临床相关性和影响;（c）它是一个多学科项目涵盖基础生物医学研究，高通量检测开发，并为未来的药物发现奠定了基础。