The circadian clock protein BMAL and post-translational regulation of ENaC in the kidney

肾脏中生物钟蛋白 BMAL 和 ENaC 的翻译后调节

• 批准号: 10662317
• 负责人: Abdel Ayube Alli
• 金额: $ 33.55万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10662317
• 关键词: ARNTL gene; Adaptor Signaling Protein; Address; Aldosterone; Antihypertensive Agents; Apical; Attenuated; Binding; Biological Assay; Biotinylation; Blood Pressure; Cardiovascular Diseases; Caspase; Cathepsins B; Cause of Death; Cell membrane; Cells; Clock protein; Data; Distal; Duct (organ) structure; Ductal Epithelial Cell; Electrolytes; Equilibrium; Essential Hypertension; Etiology; Excretory function; Exhibits; Female; Fluorescence Resonance Energy Transfer; Genes; Goals; Grant; Hour; Hypertension; In Situ; Infusion procedures; Investigation; Ion Channel; Kidney; Knockout Mice; Knowledge; MARCKS gene; Mass Spectrum Analysis; Measures; Mediating; Membrane; Metabolic; Molecular; Mus; Nephrons; Patch-Clamp Techniques; Pathogenesis; Peptide Hydrolases; Phase; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Phospholipids; Play; Post-Translational Protein Processing; Post-Translational Regulation; Potassium; Probability; Proteins; Proteolysis; Proteome; Proteomics; Regulation; Role; Sex Differences; Sodium; Telemetry; Testing; Therapeutic; Time; United States; Up-Regulation; Western Blotting; Wild Type Mouse; absorption; alpha 1-Antitrypsin; apical membrane; blood pressure control; blood pressure reduction; blood pressure regulation; cardiovascular risk factor; circadian; circadian pacemaker; circadian regulation; comparison control; density; epithelial Na+ channel; experimental study; male; new therapeutic target; novel; overexpression; protein expression; renal epithelium; salt balance; sodium channel proteins; transcription factor; urinary

ABSTRACT The epithelial sodium channel (ENaC) expressed in the distal tubule and collecting duct is responsible for the final regulation of sodium reabsorption by the kidneys. The myristoylated alanine-rich C kinase substrate (MARCKS) plays an important role as an adaptor protein between the anionic phospholipid PIP2 and ENaC. Both ENaC and MARCKS are positively regulated by the protease cathepsin B. First, our preliminary data demonstrate renal ENaC activity and MARCKS protein expression are positively regulated by the circadian protein BMAL1. Second, our preliminary data show alpha-1 antitrypsin is increased in the BMAL1 knockout mouse kidney compared to the kidney of wild-type mice sacrificed at the same time. Third, our preliminary data show alpha-1 antitrypsin is expressed in the kidney and it strongly inhibits cathepsin B activity and contributes to blood pressure regulation. In this project we will test our hypothesis that the association between renal ENaC and MARCKS, and their function at the apical plasma membrane negatively correlates with alpha-1 antitrypsin expression in a circadian dependent manner. We will perform experiments to investigate proteolysis and apical membrane expression of ENaC and MARCKS, ENaC activity, sodium handling, and blood pressure using male and female BMAL1 knockout mice, alpha-1 antitrypsin knockout mice, alpha-1 antitrypsin overexpressing mice, cathepsin B knockout mice, and wild-type control mice. The successful completion of our proposed studies for this project will reveal new mechanisms underlying the role of BMAL1 in the regulation of renal ENaC and MARCKS and blood pressure control. Our long term goal is to provide a better understanding for the pathogenesis of essential hypertension that can potentially lead to novel drug targets and therapeutics.

抽象的 上皮钠通道 (ENaC) 在远端小管和集合管中表达 负责肾脏钠重吸收的最终调节。这 肉豆蔻酰化富含丙氨酸的 C 激酶底物 (MARCKS) 作为 阴离子磷脂 PIP2 和 ENaC 之间的衔接蛋白。 ENaC 和 MARCKS 受到蛋白酶组织蛋白酶 B 的正向调节。首先，我们初步 数据表明肾 ENaC 活性和 MARCKS 蛋白表达呈阳性 受昼夜节律蛋白 BMAL1 调节。其次，我们的初步数据显示alpha-1 与正常小鼠的肾脏相比，BMAL1 敲除小鼠肾脏中的抗胰蛋白酶含量增加 野生型小鼠同时处死。三、我们的初步数据显示alpha-1 抗胰蛋白酶在肾脏中表达，强烈抑制组织蛋白酶 B 的活性， 有助于血压调节。在这个项目中，我们将测试我们的假设 肾 ENaC 和 MARCKS 之间的关联及其在心尖部的功能 质膜与 α-1 抗胰蛋白酶表达呈负相关 昼夜节律依赖方式。我们将进行实验来研究蛋白水解作用 ENaC 和 MARCKS 的顶膜表达、ENaC 活性、钠 使用雄性和雌性 BMAL1 敲除小鼠 alpha-1 进行处理和血压测量 抗胰蛋白酶敲除小鼠、α-1 抗胰蛋白酶过表达小鼠、组织蛋白酶 B 基因敲除小鼠和野生型对照小鼠。顺利完成我们提出的 该项目的研究将揭示 BMAL1 在 肾脏 ENaC 和 MARCKS 的调节以及血压控制。我们的长期目标 旨在更好地了解原发性高血压的发病机制 可能会产生新的药物靶点和治疗方法。

项目成果

Tempol treatment normalizes membrane expression of epithelial transport proteins in the kidney of salt-loaded hypertensive diabetic db/db mice.

Tempol 治疗可使盐负荷高血压糖尿病 db/db 小鼠肾脏中上皮转运蛋白的膜表达正常化。

• 发表时间: 2023
• 作者: Dogan, Yunus E;Bala, Niharika;Chacko, Kevin M;Tuna, Kubra M;Alli, Abdel A
• 通讯作者: Alli, Abdel A

Circadian clocks of the kidney: function, mechanism, and regulation.

肾脏的昼夜节律时钟：功能、机制和调节。

• 发表时间: 2022-10-01
• 影响因子: 33.6
• 作者: Costello, Hannah M;Johnston, Jermaine G;Juffre, Alexandria;Crislip, G Ryan;Gumz, Michelle L
• 通讯作者: Gumz, Michelle L

Tempol Alters Urinary Extracellular Vesicle Lipid Content and Release While Reducing Blood Pressure during the Development of Salt-Sensitive Hypertension.

Tempol 改变尿细胞外囊泡脂质含量和释放，同时在盐敏感性高血压的发展过程中降低血压。

• 发表时间: 2021-12-01
• 影响因子: 5.5
• 作者: Chacko, Kevin M;Nouri, Mohammad;Schramm, Whitney C;Malik, Zeeshan;Liu, Lauren P;Denslow, Nancy D;Alli, Abdel A
• 通讯作者: Alli, Abdel A

Lipid Profiles of Urinary Extracellular Vesicles Released during the Inactive and Active Phases of Aged Male Mice with Spontaneous Hypertension.

患有自发性高血压的老年雄性小鼠在非活动期和活动期释放的尿细胞外囊泡的脂质谱。

• 发表时间: 2022-12-06
• 影响因子: 5.6
• 作者: Lopez, Juliana Pena;Nouri, Mohammad;Ebrahim, Areej;Chacko, Kevin M;Schramm, Whitney C;Gholam, Mohammed F;Ozrazgat;Denslow, Nancy D;Alli, Abdel A
• 通讯作者: Alli, Abdel A

Transgenic Mice Overexpressing Human Alpha-1 Antitrypsin Exhibit Low Blood Pressure and Altered Epithelial Transport Mechanisms in the Inactive and Active Cycles.

过度表达人类 Alpha-1 抗胰蛋白酶的转基因小鼠表现出低血压，并改变非活动和活动周期中的上皮运输机制。

• 发表时间: 2021
• 作者: Liu, Lauren P;Gholam, Mohammed F;Elshikha, Ahmed Samir;Kawakibi, Tamim;Elmoujahid, Nasseem;Moussa, Hassan H;Song, Sihong;Alli, Abdel A
• 通讯作者: Alli, Abdel A