Role of nonmuscle myosin IIA in sex-dependent failed repair mechanisms in acute kidney injury

非肌肉肌球蛋白 IIA 在急性肾损伤性别依赖性失败修复机制中的作用

基本信息

批准号：
10311908
负责人：
Eryn E. Dixon
金额：
$ 6.87万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10311908
关键词：
Actins Acute Acute Renal Failure with Renal Papillary Necrosis Adhesions Adopted Affect Atlases Automobile Driving Bilateral Binding Binding Proteins Biochemical Biological Biological Assay Biology Cell Adhesion Molecules Cell Nucleus Cell Polarity Cell Shape Cell model Cell physiology Cells Centers for Disease Control and Prevention (U.S.)Characteristics Chronic Kidney Failure Cytokinesis Cytoskeleton Data Disease Management Disease Progression Down-Regulation Elements End stage renal failure Event Exhibits Female Fibrosis Filament Genes Genetic Markers Gold Healthcare Hospitalization Hour In Vitro Inflammation Injury Injury to Kidney Investigation Ischemia Kidney Laboratories Light Liver Lung Maps Morphology Mus Nonmuscle Myosin Type IIA Outcome Pathway interactions Patients Phenotype Phosphorylation Phosphotransferases Play Population Pre-Clinical Model Prevalence Property RNA Recovery Regulation Renal function Reperfusion Injury Reporting Role Sepsis Serine Sex Differences Signal Pathway Signal Transduction Structure System Techniques Threonine Time Tissues Toxin Translating United States Woman XCL1 gene cell motility differential expression effective therapy epidemiologic data genome wide association study improved improved outcome in vivo injured injury and repair insight ischemic injury kidney cell male men migration multimodality non-muscle myosin novel protective factors protein expression repaired response rho sex sexual dimorphism stem therapeutic target therapeutically effective transcriptome sequencing transcriptomics

项目摘要

ABSTRACT The cellular mechanisms that drive the progression of acute kidney injury (AKI) to chronic kidney disease (CKD) are not well understood. AKI is the abrupt decrease of kidney function from a variety of causes that elicits structural and functional damage. The prevalence of AKI is increasing rapidly in the United States and presents a healthcare burden worldwide. To date, there are no effective therapies for the mitigation of the proinflammatory and fibrotic phenotypes resulting from injury. Interestingly, it has been recently supported with epidemiological data that female sex is a protective factor in the AKI progression to CKD. Leveraging the comparison of sex specific mechanisms in ischemic injury has incredible potential for the identification of novel, effective therapeutic targets. Therefore, I will be employing single nuclei RNA and ATAC-Sequencing, as well as spatial transcriptomics to generate a multimodal atlas of the injury time course in females undergoing bilateral ischemia reperfusion injury (Bi-IRI). Recently, a novel cell state has been identified in male mice following Bi-IRI using single nuclei RNA sequencing. Known as failed repair proximal tubule cells (FR-PTC), these cells represent a dead end in the recovery pathway and could be lending a role to increases in proliferation, fibrosis, and inflammation that worsen kidney tissue after injury. These FR-PTCs exhibit a profile of differentially expressed genes that are unique from healthy and repairing cell states. One of these genetic markers, which is also a genome wide association study (GWAS) gene associated with CKD, was Myh9. Myh9 expression is upregulated specifically in injured and failed repair cell clusters, and is found in low levels in healthy and repairing proximal tubule cells. This gene encodes for nonmuscle myosin IIA (NMIIA), which plays an important role in cell shape, adhesion, migration, and cytokinesis. It interacts directly with actin, creating a stable cytoskeletal network. Drastic changes in Myh9 expression may translate into changes in NMIIA abundance that could alter the regulation of downstream signaling, contributing to phenotypes characteristic of AKI. Therefore, I hypothesize that sexually dimorphic expression of Myh9 in the kidney and sex-specific mechanisms of NMIIA activation contribute to the renoprotection of female sex. Preliminary data suggests that female C57BL/6J mice at baseline have higher expression of Myh9 than males, potentially indicating that females have developed mechanisms for decreasing activity of NMIIA through downregulation of ischemia responsive kinases. This proposal will investigate these expression changes in males and females along the injury time course following Bi-IRI using single nuclei sequencing and spatial transcriptomics. Additionally, the relationship between ischemia and NMIIA function will be investigated using gold standard biochemical techniques in in vivo and in vitro cell models. Ultimately, this proposal will generate a transcriptomic atlas of sex differences in AKI and illuminate the role of NMIIA in the disease progression of AKI to CKD, potentially identifying a new sex-specific therapeutic target.

抽象的驱动急性肾损伤（AKI）进展为慢性肾病（CKD）的细胞机制没有被很好地理解。 AKI 是多种原因引起的肾功能突然下降结构和功能损坏。 AKI 的患病率在美国迅速增加，并呈现出全世界的医疗保健负担。迄今为止，尚无有效的疗法来缓解促炎性炎症以及损伤引起的纤维化表型。有趣的是，它最近得到了流行病学的支持数据表明，女性是 AKI 进展为 CKD 的保护因素。利用性别比较缺血性损伤的具体机制对于识别新颖、有效的治疗方法具有令人难以置信的潜力目标。因此，我将采用单核 RNA 和 ATAC 测序，以及空间测序转录组学可生成双侧缺血女性损伤时间过程的多模式图谱再灌注损伤（Bi-IRI）。最近，使用 Bi-IRI 在雄性小鼠中发现了一种新的细胞状态单核RNA测序。这些细胞被称为近端小管修复失败细胞（FR-PTC）恢复途径中的死胡同，可能会导致增殖、纤维化和损伤后使肾组织恶化的炎症。这些 FR-PTC 表现出差异表达的特征健康和修复细胞状态所特有的基因。这些遗传标记之一，也是全基因组关联研究（GWAS）与 CKD 相关的基因是 Myh9。 Myh9 表达上调特别是在受损和失败的修复细胞簇中，并且在健康和修复的近端细胞中含量较低肾小管细胞。该基因编码非肌肉肌球蛋白 IIA (NMIIA)，它在细胞形状中发挥重要作用，粘附、迁移和胞质分裂。它直接与肌动蛋白相互作用，形成稳定的细胞骨架网络。激烈的 Myh9 表达的变化可能会转化为 NMIIA 丰度的变化，从而改变对下游信号传导，有助于形成 AKI 的表型特征。因此，我推测性行为 Myh9 在肾脏中的二态性表达和 NMIIA 激活的性别特异性机制有助于对女性性的肾脏保护。初步数据表明，雌性 C57BL/6J 小鼠在基线时具有更高的 Myh9 的表达高于男性，这可能表明女性已经形成了降低 Myh9 表达的机制。通过下调缺血反应性激酶来调节 NMIIA 的活性。本提案将调查这些使用单核 Bi-IRI 后，男性和女性在损伤时间过程中的表达变化测序和空间转录组学。此外，缺血与 NMIIA 功能之间的关系将使用金标准生化技术在体内和体外细胞模型中进行研究。最终，这该提案将生成 AKI 性别差异的转录组图谱，并阐明 NMIIA 在 AKI 中的作用 AKI 到 CKD 的疾病进展，有可能确定新的性别特异性治疗靶点。