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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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的心脏通过心肌细胞肥大生长来应对压力并发展为心力衰竭当压力持续存在时。我们之前的研究表明，肥大对多种刺激的反应发生独立于钙/钙调蛋白依赖性蛋白激酶 II (CaMKII) 的信号传导，但当 CaMKII 被删除时，心力衰竭的进展就会减弱。炎症是导致该病的一个关键因素与心力衰竭相关的不良重塑。该提案的长期目标是证明心肌细胞内的 CaMKII 信号传导可引发心脏炎症反应非缺血性干预措施，例如压力超负荷（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 CaMKII 使用心脏特异性 CaMKII 敲除小鼠 (CKO) 来证明信号传导驱动心脏炎症 TAC 诱导的炎症基因表达和炎症小体激活的丧失。我们确定这些是否通过从 CTL 中分离成年小鼠心室肌细胞和 CKO 小鼠，并通过组织切片的原位杂交和酶测定。 CaMKIIC 转基因和使用具有心脏特异性 NFkB p65 亚基 KO 的小鼠来进一步证明心肌细胞引发炎症。目标 2 询问心肌细胞 CaMKII 信号传导是否有助于炎症/免疫细胞对 TAC 的反应而积累。为了实现这一目标，我们使用 CKO 小鼠来证明 TAC 通过心肌细胞 CaMKII 启动信号促进免疫细胞反应。研究重点是巨噬细胞和 T 细胞，使用免疫组织化学和流式细胞术以及单一细胞 RNA seq 全面定义心脏中积累的特定巨噬细胞群。心脏特异性 KO 或趋化因子/细胞因子的敲低用于证明这些因子的产生心肌细胞中的介质触发特定免疫细胞类型的反应。目标 3 确定是否封锁心肌细胞 CaMKII 引发的炎症减轻了不良重塑，并且在什么时候会出现这种情况需要完成。拟议的研究使用心脏特异性 KO 或抑制选定的炎症介体证明它们在心肌细胞中的形成对于纤维化的发展至关重要 TAC 后心室功能障碍。使用 AAV9 Cre 进行条件基因删除来确定时间 CaMKII 抑制的最大益处得以实现。我们的发现应该会对未来产生重大影响由于心肌细胞以前不被认为是炎症信号的产生者，因此进行了研究，在没有“警报”的情况下炎症反应被激活的机制尚未被证实迄今为止已确定，并且最有效地预防心力衰竭发展的概念是通过早期心肌细胞靶向抗炎干预来实现这一目标是新颖的。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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