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性药物，以及法律娱乐性和 在过去的十年中，药用使用率有所增加。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。肾脏产生 内源性大麻素（EC）增加尿量，对Na+和K+排泄的影响可变 啮齿动物。在人类中，很少有研究表明大麻素可以充当利尿剂，不仅促进尿Na+ 但也是hco3-损失。我们在小鼠中最初表明CB1R在以PHI的调节中表达并损害PHI 在插入细胞（IC）中，皮质收集管（CCD）中负责H+/HCO3-分泌的细胞 和Na+（和K+）传输。我们还表明，大麻素受体激动剂急性增加 小鼠的水排泄。因此，我们假设IC参与CB1R介导的利尿 通过与主要细胞交叉对话，负责Na+和水重吸收的细胞。 该VA职业发展奖的应用程序旨在定义EC在CCD中的作用。三个具体 将研究目标（SA）以评估CB1R信号对功能的存在和物理意义 鼠标CCD。 SA1将定义EC系统组件的细胞特异性表达； SA2将使用药理学确定CB1R信号传导在CCD中特异性功能中的作用 CB1R的激活剂/抑制剂； SA3将确定CB1R在IC中的靶向缺失是否改变了细胞特异性 功能。结果有望提供有关大麻素对肾脏影响的新见解，并允许 美国确定用于开发新型治疗剂的靶标（例如CB1R激动剂） 高血压，体积超负荷，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，课程和研讨会，以增强 PI（董事会认证的肾脏科医生和认证高血压专家Joshua Rein博士）的熟练程度 单管显微解剖，分离的体外微灌注，功能性荧光显微镜，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，包括 开发转基因小鼠模型。这些技能将由导师，顾问团队加强 合作者，所有人都具有必要的专业知识，知识和精明的轨道记录。 总而言之，该VA CDA赠款提案为Rein博士提供了强大的职业发展计划 他的职业目标是发展为肾脏EC系统专家的国家声誉，旨在揭露小说 电解质和酸/碱平衡的肾脏调节的基础分子途径。他的长期目标是 将临床前研究的关键观察转化为对人类疾病和 疗法。他的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·瓦姆（Peters VAMC）和西奈山（Mount Sinai）的伊坎医学院为 Rein博士的职业发展，领导力致力于他的成功，有75％的人保护时间 研究。