Role of Adipocyte Na/K-ATPase Signaling in the Development and Progression of Uremic Cardiomyopathy By Altering Adipocyte Phenotype

脂肪细胞 Na/K-ATP 酶信号传导通过改变脂肪细胞表型在尿毒症心肌病发生和进展中的作用

• 批准号: 10046425
• 负责人: Komal Sodhi
• 金额: $ 44.4万
• 依托单位: MARSHALL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-05 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10046425
• 关键词: Address; Adipocytes; Adipose tissue; Affect; Amplifiers; Attenuated; Biochemical; Cardiac; Cardiomyopathies; Cardiovascular Diseases; Cardiovascular system; Cells; Chronic Kidney Failure; Clinical; Cresol; Data; Development; Disease; Disease model; End stage renal failure; Experimental Models; Exposure to; Functional disorder; Future; Gene Transfer; Heart; In Vitro; Indican; Inflammatory; Intervention; Kidney; Kidney Failure; Lentivirus Vector; Mediator of activation protein; Metabolic; Metabolic syndrome; Methods; Modeling; Molecular; Morbidity - disease rate; Morphology; Mus; Myocardium; N Domain; Na(+)-K(+)-Exchanging ATPase; Nephrectomy; Obesity; Outcome; Oxidants; Oxidative Stress; Pathogenesis; Pathway interactions; Pattern; Peptides; Permeability; Phenotype; Physiological; Play; Prevention; Production; Protocols documentation; Reactive Oxygen Species; Reporting; Role; SRC gene; Signal Transduction; Skeletal Muscle; Source; Stress; Subfamily lentivirinae; Testing; Time; Tissues; Toxin; Work; base; cytokine; cytokine release syndrome; dietary manipulation; experimental study; improved; in vivo; interest; molecular phenotype; mortality; mouse model; novel; oxidant stress; promoter; receptor; small hairpin RNA; therapeutic target; transcriptome sequencing; uremic cardiomyopathy; vector; western diet

The role of oxidant stress in the pathophysiology of cardiovascular disease has long been a subject of considerable interest. Although end stage renal disease is relatively uncommon, milder degrees of chronic kidney disease [CKD] such as CKD 3 are quite common and have been implicated as important determinant in cardiovascular morbidity and mortality. Oxidant stress plays a key role in the development of renal-failure- associated cardiomyopathy (also known as uremic cardiomyopathy), both experimentally and clinically. The “Na/K-ATPase,” a sodium-potassium pump, has been shown by our group to affect cellular signaling via the “Na/K-ATPase signaling,” which amplifies oxidative stress. We have specifically shown that this pathway is critical to the pathophysiology of several experimental models of disease including obesity/metabolic syndrome and experimental uremic cardiomyopathy. We and others have also observed that the adipocyte itself is an important source of oxidant stress in models with obesity/metabolic syndrome and that mediators directly tied to the cellular phenotype of these adipocytes play a causal role in the cardiovascular conditions associated with obesity/metabolic syndrome. This led us to believe that adipocytes could play a central role in uremic cardiomyopathy. Therefore, we hypothesize that adipocytes create systemic oxidant stress through the Na/K- ATPase feed-forward oxidant amplification loop in uremic cardiomyopathy and serve as a therapeutic target for this condition. Our group has developed a cell permeant peptide, NaKtide, from the N domain of the α1 subunit of the Na/K-ATPase, which inhibits Na/K-ATPase-ROS amplification. Our preliminary results show that the NaKtide, targeted specifically to adipocyte, attenuates oxidative stress and inflammatory cytokines, in addition to improving metabolic parameters. Our experimental approach includes studies to determine the role of adipocyte Na/K-ATPase signaling in the development of experimental uremic cardiomyopathy using a partial nephrectomy (PNx) mouse model with and without dietary manipulations (Aim 1). We will also target the NaKtide to the adipocyte using “lentiviral gene transfer” strategy (Aim 1) to determine the role of adipocyte-derived Na/K- ATPase/Src signaling in the progression of uremic cardiomyopathy. To test the off-target effects of NaKtide, we will also use lentiviral vectors with tissue specific promoters, including heart and kidney to targeted NaKtide specially in these tissues. Further, we will target c-Src-shRNA to adipocytes using lentivirus vector, as an alternate strategy of inhibiting Na/K-ATPase signaling, to demonstrate the role of c-Src as a downstream mediator of Na/K-ATPase in exacerbating oxidative stress and eventually uremic cardiomyopathy. In Aim 2, we will employ in vitro protocols for primary adipocytes, isolated from Sham or PNx operated C57BL6 mice and perform RNASeq analysis to study the adipocyte phenotypic alterations and pathways associated with uremic cardiomyopathy phenotype. We will also use primary adipocytes isolated from C57BL6 and Na/K-ATPase α1+/- mice which will be exposed to uremic toxins, subsequently activating Na/K-ATPase signaling, to determine the activation of specific downstream molecular pathways in vitro that mimic in vivo outcomes. These experiments will allow us to determine if the Na/K-ATPase signaling and/or adipocytes are potential targets for disease intervention. These studies if proven may provide a basis for the future studies to ameliorate uremic cardiomyopathy phenotype.

氧化应激在心血管疾病病理生理学中的作用长期以来一直是 相当大的兴趣。虽然终末期肾病相对少见，但轻度慢性肾病 疾病[CKD]如CKD 3是相当常见的，并已被认为是重要的决定因素， 心血管疾病发病率和死亡率。氧化应激在肾衰竭的发展中起着关键作用， 相关性心肌病（也称为尿毒症心肌病），实验和临床。 Na/K-ATPase是一种钠钾泵，我们的研究小组已经证明它会影响细胞信号传导 通过“Na/K-ATP酶信号”，这放大了氧化应激。我们已经明确表明， 对包括肥胖/代谢综合征在内的几种疾病实验模型的病理生理学至关重要 和实验性尿毒症心肌病我们和其他人也观察到脂肪细胞本身是一种 肥胖/代谢综合征模型中氧化应激重要来源和与之直接相关的介质 这些脂肪细胞的细胞表型在心血管疾病中起因果作用 肥胖/代谢综合征。这使我们相信脂肪细胞在尿毒症中起着重要作用。 心肌病因此，我们假设脂肪细胞通过Na/K- 尿毒症心肌病中ATP酶前馈氧化放大环及其作为治疗靶点的作用 这个条件。本课题组从α1亚基的N结构域开发了一种细胞渗透肽NaKtide 抑制Na/K-ATP酶-ROS扩增。我们的初步结果表明， NaKtide，特别针对脂肪细胞，减弱氧化应激和炎症细胞因子，此外， 改善代谢参数我们的实验方法包括研究以确定 使用部分脂肪细胞Na/K-ATP酶信号在实验性尿毒症心肌病发生中的作用 肾切除术（PNx）小鼠模型，有和没有饮食操作（目的1）。我们还将瞄准NaKtide 使用“慢病毒基因转移”策略（目的1）向脂肪细胞转移，以确定脂肪细胞来源的Na/K- ATP酶/Src信号在尿毒症心肌病进展中的作用为了测试NaKtide的脱靶效应，我们 还将使用具有组织特异性启动子的慢病毒载体，包括心脏和肾脏，以靶向NaKtide 特别是在这些组织中。此外，我们将使用慢病毒载体将c-Src-shRNA靶向脂肪细胞， 抑制Na/K-ATP酶信号传导的替代策略，以证明c-Src作为下游酶的作用。 Na/K-ATP酶介导的氧化应激加剧，最终尿毒症心肌病。在目标2中， 将采用从假手术或PNx操作的C57 BL 6小鼠分离的原代脂肪细胞的体外方案， 进行RNASeq分析以研究与尿毒症相关的脂肪细胞表型改变和途径。 心肌病表型我们还将使用从C57 BL 6和Na/K-ATP酶α1+/-分离的原代脂肪细胞 将小鼠暴露于尿毒症毒素，随后激活Na/K-ATP酶信号传导，以确定 在体外激活特异性下游分子途径，模拟体内结果。这些实验 将使我们能够确定Na/K-ATP酶信号传导和/或脂肪细胞是否是疾病的潜在靶点 干预这些研究结果为进一步改善尿毒症的研究提供了依据 心肌病表型

项目成果

Na/K-ATPase suppresses LPS-induced pro-inflammatory signaling through Lyn.

• DOI: 10.1016/j.isci.2022.104963
• 发表时间: 2022-09-16
• 期刊: ISCIENCE
• 影响因子: 5.8
• 作者: Zhang, Jue;Chang, Jackie;Beg, Mirza Ahmar;Huang, Wenxin;Zhao, Yiqiong;Dai, Wen;Wu, Xiaopeng;Cui, Weiguo;Pillai, Sneha S.;Lakhani, Hari Vishal;Sodhi, Komal;Shapiro, Joseph I.;Sahoo, Daisy;Zheng, Ze;Silverstein, Roy L.;Chen, Yiliang
• 通讯作者: Chen, Yiliang

Pivotal role of adipocyte-Na/K-ATPase Signaling in the pathogenesis of Experimental Uremic Cardiomyopathy by fat transplantation.

脂肪细胞-Na/K-ATP酶信号传导在脂肪移植引起的实验性尿毒症心肌病发病机制中的关键作用。

• 发表时间: 2022
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Pillai,SnehaS;Lakhani,HariVishal;Zehra,Mishghan;Pereira,DuaneG;Sodhi,Komal
• 通讯作者: Sodhi,Komal

Mechanisms in Chronic Kidney Disease and Associated Cardiovascular Remodeling Contributes to the Progression of Cognitive Impairment in Women.

慢性肾脏病和相关心血管重塑的机制导致女性认知障碍的进展。

• 发表时间: 2022
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Lakhani,HariVishal;Thakur,Amrit;Pillai,SnehaS;Pereira,DuaneG;Thompson,Ellen;Sodhi,Komal;Fedorova,OlgaV
• 通讯作者: Fedorova,OlgaV

The Na/K-ATPase Signaling and SGLT2 Inhibitor-Mediated Cardiorenal Protection: A Crossed Road?

• DOI: 10.1007/s00232-021-00192-z
• 发表时间: 2021-12
• 期刊: The Journal of membrane biology
• 作者: Liu J;Tian J;Sodhi K;Shapiro JI
• 通讯作者: Shapiro JI

Optimization of Prescription Drug Monitoring Program to Overcome Opioid Epidemic in West Virginia.

• DOI: 10.7759/cureus.22434
• 发表时间: 2022-03
• 期刊: Cureus
• 作者: Al-Astal AY;Sodhi K;Lakhani HV
• 通讯作者: Lakhani HV