Regulation of myocardial GPCRs by USP20 in normal and hypertrophied heart

USP20 对正常和肥厚心脏中心肌 GPCR 的调节

• 批准号: 10630331
• 负责人: SUDHA K SHENOY
• 金额: $ 56.21万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10630331
• 关键词: Address; Affect; Affinity; Agonist; American; Apoptosis; Autophagocytosis; Autophagosome; Biomechanics; Blood Pressure; C57BL/6 Mouse; Cardiac; Cardiac Myocytes; Cardiomegaly; Cellular Stress; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Compensation; Complex; Control Groups; Coupling; Cyclic AMP-Dependent Protein Kinases; Deubiquitination; Development; Down-Regulation; Drug Screening; Fibrosis; Functional disorder; G-Protein-Coupled Receptors; GTP-Binding Proteins; Genes; Heart; Heart Hypertrophy; Heart failure; Human; Hypertension; Hypertrophy; Isoproterenol; Knock-in Mouse; Knockout Mice; Label; Left Ventricular Hypertrophy; Left Ventricular Remodeling; Ligands; Link; LoxP-flanked allele; Lysine; Mass Spectrum Analysis; Mediating; Mus; Myocardial; Myocardial dysfunction; Myocardium; Pathologic; Pathway interactions; Patients; Phosphorylation; Phosphorylation Inhibition; Play; Polyubiquitination; Proteins; Receptor Activation; Regulation; Research; Role; Signal Induction; Signal Transduction; Testing; Transducers; Ubiquitination; aorta constriction; beta-2 Adrenergic Receptors; cardioprotection; desensitization; drug discovery; experimental study; heart function; hypertensive; in vivo; mortality; novel; pressure; prevent; protein expression; receptor; response; trafficking; ubiquitin isopeptidase; ubiquitin-specific protease

PROJECT SUMMARY Chronic pressure overload that leads to left ventricular hypertrophy (LVH) and adverse cardiac remodeling is one of the leading causes of heart failure and affects millions of Americans. In addition to pathological insults and biomechanical factors, neurohormonal pathways that involve coordinated signaling through β1 and β2 adrenergic receptors (ARs) can play a major role in myocardial adaptation to pressure overload. The factors that shift the myocardial response to pressure overload from adaptive to maladaptive LVH remain largely obscure. Our research has focused on the regulation of βAR trafficking and signaling by ubiquitination/deubiquitination mechanisms. We recently discovered that differential regulation of β1AR and β2AR endocytic trafficking can be achieved by the deubiquitinase (DUB) called ubiquitin-specific protease-20 (USP20), which we found deubiquitinates both βARs. In response to the βAR agonist (-)isoproterenol (Iso), USP20 is phosphorylated on Ser333 by protein kinase A (PKA); this phosphorylation inhibits USP20 DUB activity toward the β2AR and regulates trafficking of the β2AR to autophagosomes. USP20 phosphorylation occurs in vivo, as well: (1) USP20 phosphorylation is significantly elevated in failing human hearts when compared with non-failing hearts and (2) pressure overload achieved by transverse aortic constriction (TAC) triggers USP20 phosphorylation in cardiomyocytes of WT, but not β1AR KO mice. Accordingly, during LVH, USP20 phosphorylation requires β1AR activation in the heart and may play a role in pathologic remodeling in LVH and/or heart failure. We hypothesize that “USP20 and its phosphorylation status fine-tune βAR signal transduction, endocytic trafficking and autophagy, thus impacting cardiac remodeling in LVH.” We will test our hypotheses by addressing the following specific aims: (1) To determine the role of cardiomyocyte-USP20 in the development of cardiac dysfunction, (2) To determine the regulation of myocardial βAR signaling by USP20 Ser333 phosphorylation and (3) To determine the coordinated roles of β1AR and USP20 in regulating ubiquitination status of the autophagy protein Beclin1 and its effect in cardiomyocyte autophagy.

项目摘要 慢性压力超负荷，导致左心室肥大（LVH）和不良心脏重塑 是一个一个一个一个一个一个一个一个一个心力衰竭的一个一个一个，并且是asea术 侮辱和生物力学因子，涉及通过β1和 β2肾上腺素能受体（ARS）可以在压力超负荷的心肌适应中起主要作用 这将心肌反应转移到压力超负荷从自适应到适应不良的LVH仍然很大程度上保持 我们的研究晦涩。 泛素化/去泛素化机制。 β2AR内吞运输可以通过称为泛素特异性蛋白酶-20的去泛素酶（DUB）来实现 （USP20），我们发现对βAR激动剂（ - ）异丙肾上腺素（ISO），deubiqueItin USP20在Ser333上被蛋白激酶A（PKA）抑制了USP20 DUB； 对β2AR的活性和β2AR的规则运输到自噬体 也发生在体内：（1）在失败的心脏中，USP20磷酸化显着升高 与非释放心脏和（2）通过横向主动脉结构（TAC）实现的压力超负荷相比 触发WT的心肌细胞中的USP20磷酸化，但没有β1ARKO小鼠。 USP20磷酸化需要心脏中的β1AR激活，并可能在病理重塑中发挥作用 LVH和心力衰竭。 信号转导，内吞运输和自噬，从而影响LVH的心脏重塑。 我们将通过解决以下特定目的来检验我们的假设：（1）确定 心脏功能障碍发展中心肌细胞-USP20，（2）确定调节 USP20 Ser333磷酸化和（3）确定协调的心肌βAR信号传导 β1AR和USP20在调节自噬蛋白11及其ITS的泛素化状态中的作用 心肌细胞自噬的作用。

项目成果

Editorial: Regulation of hormone and growth factor signalling by ubiquitin and ubiquitin-like protein modifications.

社论：通过泛素和泛素样蛋白修饰调节激素和生长因子信号传导。

• DOI: 10.3389/fendo.2024.1397685
• 发表时间: 2024
• 期刊: Frontiers in endocrinology
• 影响因子: 5.2
• 作者: Shenoy,SudhaK;Grimsey,NeilJ;Piper,RobertC
• 通讯作者: Piper,RobertC

Agonist-activated glucagon receptors are deubiquitinated at early endosomes by two distinct deubiquitinases to facilitate Rab4a-dependent recycling.

• DOI: 10.1074/jbc.ra120.014532
• 发表时间: 2020-12-04
• 期刊: The Journal of biological chemistry
• 作者: Kaur S;Chen Y;Shenoy SK
• 通讯作者: Shenoy SK

The ubiquitination status of the glucagon receptor determines signal bias.

• DOI: 10.1016/j.jbc.2023.104690
• 发表时间: 2023-05
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Kaur, Suneet;Sokrat, Badr;Capozzi, Megan E.;El, Kimberley;Bai, Yushi;Jazic, Aeva;Han, Bridgette;Kumar, Kaavya Krishna;D'Alessio, David A.;Campbell, Jonathan E.;Bouvier, Michel;Shenoy, Sudha K.
• 通讯作者: Shenoy, Sudha K.

A single phenylalanine residue in β-arrestin2 critically regulates its binding to G protein-coupled receptors.

• DOI: 10.1016/j.jbc.2022.101837
• 发表时间: 2022-05
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Jean-Charles, Pierre-Yves;Rajiv, Vishwaesh;Sarker, Subhodeep;Han, Sangoh;Bai, Yushi;Masoudi, Ali;Shenoy, Sudha K.
• 通讯作者: Shenoy, Sudha K.