Cardiomyocyte CaM kinase II as a driver of cardiac inflammation and remodeling

心肌细胞 CaM 激酶 II 作为心脏炎症和重塑的驱动因素

• 批准号: 10308392
• 负责人: JOAN HELLER BROWN
• 金额: $ 39.5万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-15 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10308392
• 关键词: Adult; Anti-Inflammatory Agents; Attenuated; Biological Assay; CCL2 gene; Ca(2+)-Calmodulin Dependent Protein Kinase; Cardiac; Cardiac Myocytes; Cause of Death; Cells; Development; Failure; Fibrosis; Flow Cytometry; Functional disorder; Gene Deletion; Gene Expression; Generations; Genetic Transcription; Goals; Growth; Heart; Heart Diseases; Heart failure; Hypertrophy; Immune; Immunohistochemistry; In Situ Hybridization; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Interleukin-1; Interleukin-18; Intervention; Knock-out; Knockout Mice; Lead; Mediating; Mediator of activation protein; Mus; Muscle Cells; Myocarditis; Necrosis; Phenotype; Play; Population; Prevention; Process; Role; Signal Transduction; Signaling Molecule; Site; Stimulus; Stress; T-Lymphocyte; Time; Tissues; Transgenic Mice; Transgenic Organisms; Ventricular; Ventricular Dysfunction; Work; calmodulin-dependent protein kinase II; cardiogenesis; cell type; chemokine; cytokine; immune activation; knock-down; macrophage; novel; p65; pressure; prevent; recruit; response; single-cell RNA sequencing

ABSTRACT The heart responds to stress through hypertrophic growth of cardiomyocytes and progresses to heart failure when stress is sustained. Our previous studies showed that hypertrophy in response to a variety of stimuli occurs independent of signaling through the calcium/calmodulin dependent protein kinase II (CaMKII) but that progression to heart failure is attenuated when CaMKII is deleted. Inflammation is a key contributor to the adverse remodeling associated with heart failure. The long term objective of this proposal is to demonstrate that CaMKII signaling within cardiomyocytes initiates cardiac inflammation in response to non-ischemic interventions such as pressure overload (TAC) and that this plays a significant role in the development of heart failure. Studies proposed in Aim 1 determine if cardiomyocyte localized CaMKII signaling drives cardiac inflammation using cardiac specific CaMKII knockout mice (CKO) to demonstrate loss of TAC-induced inflammatory gene expression and inflammasome activation. We determine if these responses occur specifically in cardiomyocytes by isolation of adult mouse ventricular myocytes from CTL and CKO mice, and by in situ hybridization and enzymatic assays in tissue sections. CaMKIIC transgenics and mice with cardiac specific KO of the p65 subunit of NFkB are used to further demonstrate involvement of the cardiomyocyte in igniting inflammation. Aim 2 asks whether cardiomyocyte CaMKII signaling contributes to accumulation of inflammatory/immune cells in response to TAC. Work in this aim uses CKO mice to demonstrate that TAC promotes immune cell responses through cardiomyocyte CaMKII initiated signals. Studies focus on macrophages and T-cells, using immunohistochemistry and flow cytometry as well as single cell RNA seq to comprehensively define specific populations of macrophages that accumulate in the heart. Cardiac specific KOs or knockdown of chemokines/cytokines is used to demonstrate that generation of these mediators in cardiomyocytes triggers responses of specific immune cell types. Aim 3 determines if blockade of cardiomyocyte CaMKII-initiated inflammation attenuates adverse remodeling and at what point this needs to be accomplished. Proposed studies use cardiac specific KO or inhibition of selected inflammatory mediators to demonstrate that their formation in cardiomyocytes is critical for development of fibrosis and ventricular dysfunction following TAC. Conditional gene deletion with AAV9 Cre is used to establish the time at which maximal benefit from CaMKII inhibition is achieved. Our findings should significantly impact future research since the cardiomyocyte has not previously been considered as a generator of inflammatory signals, the mechanisms by which inflammatory responses are activated in the absence of “alarmins” has not heretofore been determined, and the concept that most effective prevention of heart failure development could be achieved by early cardiomyocyte-targeted anti-inflammatory interventions is novel.

摘要 心脏通过心肌细胞的肥大生长对应激作出反应，并发展为心力衰竭 当压力持续时。我们以前的研究表明，肥大反应于各种刺激， 不依赖于通过钙/钙调蛋白依赖性蛋白激酶II（CaMKII）的信号传导， 当CaMKII缺失时，心力衰竭的进展减弱。炎症是一个关键因素， 与心力衰竭相关的不良重塑。这项建议的长远目标是 证明心肌细胞内的CaMKII信号传导启动心脏炎症反应， 非缺血性干预，如压力超负荷（TAC），这在 心力衰竭的发展。目标1中提出的研究确定心肌细胞是否定位于CaMKII 使用心脏特异性CaMKII敲除小鼠（CKO）， TAC诱导的炎性基因表达和炎性小体活化的丧失。我们确定这些 通过从CTL中分离成年小鼠心室肌细胞， CKO小鼠，并通过原位杂交和组织切片中的酶测定。CaMKII β C转基因和 使用具有NFkB的p65亚基的心脏特异性KO的小鼠来进一步证明NFkB的p65亚基的参与。 心肌细胞引发炎症。目的2询问心肌细胞CaMKII信号传导是否有助于 炎症/免疫细胞响应于TAC的积聚。这项工作使用CKO小鼠， 证明TAC通过心肌细胞CaMKII启动的信号促进免疫细胞应答。 研究集中在巨噬细胞和T细胞，使用免疫组织化学和流式细胞术以及单克隆抗体。 cell RNA seq来全面定义在心脏中积聚的巨噬细胞的特定群体。 心脏特异性科斯或趋化因子/细胞因子的敲低用于证明这些细胞因子的产生 心肌细胞中的介质触发特定免疫细胞类型的应答。目标3确定是否封锁 心肌细胞CaMKII启动的炎症减弱了不良重塑， 需要完成。拟定的研究使用心脏特异性KO或抑制选定的炎性 介质，以证明它们在心肌细胞中的形成对纤维化的发展至关重要， TAC后的心室功能障碍。使用AAV 9 Cre的条件性基因缺失来确定在第一阶段的时间。 从而获得CaMKII抑制的最大益处。我们的发现将对未来产生重大影响。 研究由于心肌细胞以前没有被认为是炎症信号的发生器， 在缺乏“警报素”的情况下，炎症反应被激活的机制没有 迄今为止已经确定，最有效的预防心力衰竭发展的概念可以 通过早期心肌细胞靶向抗炎干预来实现是新颖的。

项目成果

Molecular Signaling to Preserve Mitochondrial Integrity against Ischemic Stress in the Heart: Rescue or Remove Mitochondria in Danger.

• DOI: 10.3390/cells10123330
• 发表时间: 2021-11-27
• 期刊: Cells
• 影响因子: 6
• 作者: Yu JD;Miyamoto S
• 通讯作者: Miyamoto S

SiglecF(HI) Marks Late-Stage Neutrophils of the Infarcted Heart: A Single-Cell Transcriptomic Analysis of Neutrophil Diversification.

• DOI: 10.1161/jaha.120.019019
• 发表时间: 2021-02-16
• 期刊: Journal of the American Heart Association
• 影响因子: 5.4
• 作者: Calcagno DM;Zhang C;Toomu A;Huang K;Ninh VK;Miyamoto S;Aguirre AD;Fu Z;Heller Brown J;King KR
• 通讯作者: King KR

RhoA signaling increases mitophagy and protects cardiomyocytes against ischemia by stabilizing PINK1 protein and recruiting Parkin to mitochondria.

• DOI: 10.1038/s41418-022-01032-w
• 发表时间: 2022-12
• 期刊: CELL DEATH AND DIFFERENTIATION
• 影响因子: 12.4
• 作者: Tu, MichelleZ;Tan, Valerie P.;Yu, Justin D.;Tripathi, Raghav;Bigham, Zahna;Barlow, Melissa;Smith, Jeffrey M.;Brown, Joan Heller;Miyamoto, Shigeki
• 通讯作者: Miyamoto, Shigeki

The contribution of the cardiomyocyte to tissue inflammation in cardiomyopathies.

• DOI: 10.1016/j.cophys.2020.10.003
• 发表时间: 2021-03
• 期刊: Current opinion in physiology
• 影响因子: 2.5
• 作者: Ninh VK;Brown JH
• 通讯作者: Brown JH

A Kinase Interacting Protein 1 (AKIP1) promotes cardiomyocyte elongation and physiological cardiac remodelling.

• DOI: 10.1038/s41598-023-30514-1
• 发表时间: 2023-03-10
• 期刊: SCIENTIFIC REPORTS
• 影响因子: 4.6
• 作者: Nijholt, Kirsten T.;Sanchez-Aguilera, Pablo I.;Booij, Harmen G.;Oberdorf-Maass, Silke U.;Dokter, Martin M.;Wolters, Anouk H. G.;Giepmans, Ben N. G.;van Gilst, Wiek H.;Brown, Joan H.;de Boer, Rudolf A.;Sillje, Herman H. W.;Westenbrink, B. Daan
• 通讯作者: Westenbrink, B. Daan