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

• 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
• 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

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 驱动急性肾脏损伤（AKI）发展为慢性肾脏疾病（CKD）的细胞机制 不太了解。 AKI是引起各种原因的肾功能突然下降 结构和功能损害。在美国，AKI的流行速度正在迅速增加，并呈现 全世界的医疗保健负担。迄今为止，尚无有效的减轻促炎性疗法 和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向CKD发展的保护因素。利用性比较 缺血性损伤中的特定机制具有鉴定新型，有效治疗的难以置信的潜力 目标。因此，我将使用单核RNA和ATAC-STERECTING，以及空间 转录组学以产生双侧缺血的女性伤害时间课程的多模式地图集 再灌注损伤（BI-IRI）。最近，在BI-IRI使用BI-IRI之后，在雄性小鼠中发现了一种新的细胞状态 单核RNA测序。这些细胞称为失败的修复近端小管细胞（FR-PTC） 恢复途径中的死胡同，可能会在增殖，纤维化和 受伤后肾脏组织恶化的炎症。这些FR-PTC表现出差异表达的轮廓 来自健康和修复细胞状态的独特基因。这些遗传标记之一，也是 基因组广泛的关联研究（GWAS）基因与CKD相关，是MYH9。 myh9表达被上调 特别是在受伤和失败的修复细胞簇中，在健康和修复的较低水平中发现 小管细胞。该基因编码非肌肉肌球蛋白IIA（NMIIA），该基因在细胞形状中起重要作用， 粘附，迁移和细胞因子。它与肌动蛋白直接相互作用，创建一个稳定的细胞骨架网络。剧烈 MYH9表达的变化可能转化为NMIIA丰度的变化，可能会改变调节的调节 下游信号传导，有助于AKI的表型。因此，我假设这是性的 MyH9在NMIIA激活的肾脏和性别特异性机制中的双态表达有助于 女性重新保护。初步数据表明，基线时雌性C57BL/6J小鼠的小鼠较高 MYH9的表达比男性的表达可能表明女性已经开发了减少的机制 NMIIA通过下调缺血反应激酶的活性。该提议将调查这些 BI-IRI使用单核后，男性和女性的表达变化 测序和空间转录组学。此外，缺血和NMIIA功能之间的关系将 可以在体内和体外细胞模型中使用金标准生化技术研究。最终，这个 提案将在AKI中产生性别差异的转录组，并阐明NMIIA在 AKI向CKD的疾病进展，有可能识别出新的性别特异性治疗靶点。