The regulation of renal tubular transport by cannabinoid receptor type 1 (CB1R) and its endogenous lipid ligands

1型大麻素受体（CB1R）及其内源性脂质配体对肾小管转运的调节

• 批准号: 10588113
• 负责人: Joshua L Rein
• 金额: --
• 依托单位: JAMES J PETERS VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10588113
• 关键词: 2-arachidonylglycerol; Acids; Acute; Address; Adult; Affinity; Agonist; Aldosterone; Alkalies; American; Animal Model; Apical; Applications Grants; Arachidonic Acids; Bicarbonates; Biological; Board Certification; CNR1 gene; CNR2 gene; Cannabinoids; Cannabis; Cell Separation; Cell physiology; Cells; Cellular biology; Certification; Chronic Disease; Chronic Kidney Failure; Consumption; Coupled; Development; Development Plans; Dinoprostone; Disease; Distal; Diuresis; Diuretics; Duct (organ) structure; Educational workshop; Electrolytes; Electrophysiology (science); Elements; Endocannabinoids; Environment; Enzymes; Equilibrium; Esters; Excretory function; Exhibits; Exposure to; Fluid Balance; GLUT-2 protein; GTP-Binding Proteins; Genetic; Glucose; Goals; Homeostasis; Human; Hypertension; Hyponatremia; Impairment; In Vitro; Ingestion; Intercalated Cell; Investigation; Ion Transport; K ATPase; K-Series Research Career Programs; Kidney; Kidney Diseases; Knockout Mice; Knowledge; Leadership; Legal; Ligands; Limb structure; Lipid Biochemistry; Lipids; Liquid Chromatography; Measures; Mediating; Medicine; Mentors; Mentorship; Metabolic Pathway; Metabolic acidosis; Microdissection; Microscopy; Molecular; Monoacylglycerol Lipases; Mus; NAPE-PLD; Nephrons; Oryctolagus cuniculus; Osmosis; Output; Paracrine Communication; Pathway interactions; Perfusion; Persons; Pharmacology; Physiological; Physiology; Play; Population; Potassium; Property; Proteins; Proteinuria; Rattus; Receptor Activation; Receptor Signaling; Records; Recovery; Recreation; Recreational Drugs; Regulation; Renal Blood Flow; Reporting; Research; Rodent; Role; Signal Transduction; Sodium; Sodium Bicarbonate; Sodium Chloride; Sorting; Specialist; Testing; Therapeutic; Therapeutic Agents; Thick; Time; Training; Transcript; Transgenic Mice; Translating; Tubular formation; Urine; Veterans; Water; absorption; aged; anandamide; autocrine; base; cannabinoid receptor; career; career development; cyclooxygenase 2; endogenous cannabinoid system; fatty acid amide hydrolase; fetal; human disease; inhibitor; insight; kidney epithelial cell; kidney fibrosis; lipoprotein lipase; marijuana use; medical schools; mouse model; novel; novel therapeutics; paracrine; pharmacologic; phytocannabinoid; preclinical study; renal tubular transport; response; skills; success; tandem mass spectrometry; urinary; volume hypertension

PROJECT SUMMARY/ABSTRACT: Cannabis is the most commonly used federally illicit recreational drug in the U.S., and legal recreational and medicinal use has increased over the last decade. Cannabis contains phytocannabinoids with differing affinities for the ubiquitously expressed G-protein coupled cannabinoid receptors CB1R and CB2R. The kidney produces endogenous cannabinoids (ECs) that increase urine output with variable effects on Na+ and K+ excretion in rodents. Few studies in humans suggest that cannabinoids can act as diuretics, promoting not only urinary Na+ but alsoHCO3- losses. We have preliminarily shown in mice that CB1R is expressed in and impairs pHi regulation in intercalated cells (ICs), cells in the cortical collecting duct (CCD) that are responsible for H+/HCO3- secretion and Na+ (and K+) transport. We have also shown that cannabinoid receptor agonists acutely increase urinary water excretion in mice. Therefore, we hypothesize that ICs participate in CB1R mediated diuresis possibly through cross-talk with principal cells, cells responsible for the reabsorption of Na+ and water. This VA Career Development Award application aims to define the role of ECs in the CCD. Three Specific Aims (SA) will be studied to evaluate the presence and physiological significance of CB1R signaling on function of the mouse CCD. SA1 will define cell-specific expression of components of the EC system in this segment; SA2 will determine the role of CB1R signaling on cell-specific function in the CCD, utilizing pharmacologic activators/inhibitors of CB1R; and SA3 will determine whether targeted deletion of CB1R in ICs alters cell-specific functions. The results promise to provide new insight into the effects of cannabinoids on the kidney and allows us to identify targets (e.g., CB1R agonists) for development of novel therapeutic agents for the treatment of disorders such as hypertension, volume overload, metabolic acidosis, and hyponatremia. The proposed training plan includes career development activities, courses, and workshops to enhance the proficiency of the PI (Dr. Joshua Rein, a board-certified nephrologist and certified hypertension specialist) in single tubule microdissection, isolated tubule in vitro microperfusion, functional fluorescent microscopy, lipid biochemistry and pharmacology, cell biology, electrophysiology, cell sorting, and animal models including the development of transgenic mouse models. These skills will be reinforced by a team of mentors, advisors, and collaborators, all of whom have the requisite expertise, knowledge, and mentorship track-records. In sum, this VA CDA grant proposal provides a robust career development plan for Dr. Rein to accomplish his career goals to develop a national reputation as an expert in the renal EC system, aiming to uncover novel molecular pathways underlying the renal regulation of electrolyte and acid/base balance. His long-term goal is to translate key observations from preclinical studies into an enhanced understanding of human disease and therapeutics. The expertise and support of his mentors will be critical to his career development. The James J. Peters VAMC and the Icahn School of Medicine at Mount Sinai provide outstanding academic environments for Dr. Rein’s career development, with leadership committed to his success and 75% protected time conduct his research.

项目摘要/摘要： 大麻是美国最常用的联邦非法娱乐毒品，合法的娱乐和 在过去的十年里，药用用途有所增加。大麻含有不同亲和力的植物大麻素 对于普遍表达的G蛋白偶联大麻素受体CB1R和CB2R。肾生血 内源性大麻素(ECs)增加尿量，对Na+和K+排泄有不同影响 啮齿动物。很少有人体研究表明大麻素可以起到利尿剂的作用，不仅能促进尿钠 但也有HCO3的损失。我们已经在小鼠身上初步表明，CB1R在小鼠体内表达并损害phi调节 在间质细胞(ICs)中，负责H+/HCO3-分泌的皮质集合管(CCD)中的细胞 Na+(和K+)转运。我们还发现大麻素受体激动剂能显著增加尿量。 小鼠体内的水分排泄。因此，我们推测ICs可能参与了CB1R介导的利尿。 通过与主细胞的串扰，主细胞负责对Na+和水的重吸收。 这项退伍军人职业发展奖申请旨在定义欧洲经济合作与发展组织在社区发展计划中的角色。三个具体的 将研究AIMS(SA)以评估CB1R信号在功能上的存在和生理意义 鼠标的电荷耦合器件。SA1将在这一细分中定义EC系统组件的特定细胞表达； SA2将利用药理学手段确定CB1R信号在细胞特异性功能中的作用 CB1R的激活物/抑制物；和SA3将决定ICs中CB1R的靶向缺失是否会改变细胞特异性 功能。这一结果有望为大麻类药物对肾脏的影响提供新的见解，并允许 美国将确定靶点(例如，CB1R激动剂)，以开发治疗糖尿病的新治疗剂 高血压、容量超负荷、代谢性酸中毒和低钠血症等疾病。 拟议的培训计划包括职业发展活动、课程和讲习班，以加强 专科医生(注册肾病专家和高血压专家Joshua Rein博士)在 单管显微解剖、体外肾小管微灌流、功能荧光显微镜、脂质 生物化学和药理学、细胞生物学、电生理学、细胞分选和动物模型 转基因小鼠模型的建立。这些技能将通过一个由导师、顾问和 合作者，所有人都有必要的专业知识和指导记录。 总而言之，这项退伍军人事务部CDA拨款提案为雷小山博士提供了一个强大的职业发展计划来完成 他的职业目标是在全国树立肾脏EC系统专家的声誉，旨在发现 肾脏调节电解质和酸碱平衡的分子途径。他的长期目标是 将临床前研究中的关键观察结果转化为对人类疾病的更好理解 治疗学。他导师的专业知识和支持将对他的职业发展至关重要。詹姆斯·J。 彼得斯VAMC和位于西奈山的伊坎医学院为 雷小山博士的职业发展，领导层致力于他的成功，75%的时间保护他的行为 研究。