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.

摘要 驱动急性肾损伤（阿基）进展为慢性肾病（CKD）的细胞机制 并没有得到很好的理解。阿基是由各种原因引起的肾功能突然下降， 结构性和功能性损害。阿基的患病率在美国迅速增加， 全世界的医疗负担。到目前为止，还没有有效的治疗方法来缓解促炎性疾病。 和损伤导致的纤维化表型。有趣的是，它最近得到了流行病学的支持， 数据表明，女性是阿基进展为CKD的保护因素。利用性别比较 缺血性损伤的特异性机制对于鉴定新的、有效的治疗药物具有令人难以置信的潜力。 目标的因此，我将采用单核RNA和ATAC测序，以及空间 转录组学生成双侧缺血女性损伤时间进程的多模式图谱 再灌注损伤（Bi-IRI）。最近，一种新的细胞状态已被确定在雄性小鼠后，Bi-IRI使用 单核RNA测序。这些细胞被称为修复失败的近端小管细胞（FR-PTC），它们代表了一种新的细胞。 恢复途径中的死胡同，并可能导致增殖、纤维化和纤维化的增加 损伤后恶化肾组织炎症。这些FR-PTC表现出差异表达的 健康和修复细胞状态的独特基因。这些遗传标记之一，也是一个 在全基因组关联研究（GWAS）中，与CKD相关的基因是Myh 9。Myh 9表达上调 特别是在受损和修复失败的细胞簇中，并且在健康和修复近端细胞中以低水平存在。 肾小管细胞该基因编码非肌肉肌球蛋白IIA（NMIIA），其在细胞形状中起重要作用， 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 Myh 9表达的变化可能会转化为NMIIA丰度的变化，这可能会改变Myh 9表达的调控。 下游信号传导，有助于阿基的表型特征。因此，我假设， Myh 9在肾脏中的双态表达和NMIIA激活的性别特异性机制有助于 女性肾脏保护。初步数据表明，雌性C57 BL/6 J小鼠在基线时具有更高的 Myh 9的表达高于男性，这可能表明女性已经发展出降低Myh 9表达的机制。 通过下调缺血反应性激酶抑制NMIIA活性。本提案将调查这些 雄性和雌性沿着损伤时间进程的表达变化 测序和空间转录组学。此外，缺血和NMIIA功能之间的关系将 使用金标准生物化学技术在体内和体外细胞模型中进行研究。最终这 该提案将产生阿基性别差异的转录组图谱，并阐明NMIIA在AKI中的作用。 阿基向CKD的疾病进展，可能确定新的性别特异性治疗靶点。