Immune Modulation in Hypertension

项目总结 高血压刺激激活先天和获得性免疫细胞，然后作用于靶器官，如 肾脏和血管导致组织损伤/炎症和高血压末梢器官损害。我们和其他人 已经证明在啮齿动物模型中耗尽免疫细胞亚群或单个细胞因子确实具有保护作用 对抗高血压的这些有害影响。然而，目前还没有免疫调节疗法来治疗 高血压，它影响着全世界近50%的成年人。其中一个主要原因是免疫抑制 治疗会使很大比例的人容易受到感染和恶性肿瘤的影响。最后一个共同之处 实验性和人类高血压的免疫激活途径是T细胞亚群向 促炎症T辅助17(Th17)细胞，分泌IL-17A(IL-17A)和IL-21，并远离抗- 炎症调节性T细胞(Treg)。有新的证据表明，Rho相关蛋白激酶2 (ROCK2)在辅助性T细胞中起到分子开关的作用，通过促进Th17分化，通过磷酸化 STAT3和IRF4，抑制Treg分化。在这个提议中，我们将检验新的假设 高血压与T细胞中ROCK2/STAT3/IRF4通路的激活有关，这种激活导致 Th17/Treg比值升高，加重高血压和炎症损伤。此外，我们建议 选择性抑制ROCK2将恢复内稳态T细胞平衡，而不会导致全局 免疫抑制。一种新的口服生物利用型选择性ROCK2抑制剂KD025已经完成2/3阶段 IL-17A/IL-21介导的自身免疫性疾病的临床试验 感染风险增加。我们有新的初步数据，KD025显著降低了BP并增加了 肾脏Treg/Th17细胞比率对实验性高血压的反应。在本提案的目标1中，我们将 使用可诱导的条件性ROCK2缺陷小鼠来确定ROCK2在高血压中的细胞特异性作用。至 确定T细胞ROCK2缺失是否以最小的影响恢复高血压患者的Th17/Treg平衡 对于其他类型的细胞，我们将使用一种新的单细胞技术，称为CITE-SEQ，来对循环白细胞进行表型 以及使用系统性脂多糖(LPS)模型执行免疫功能的功能分析 发炎。在目标2中，我们将确定KD025对血压、Th17/Treg平衡、终末器官损伤、 以及内毒素引起的炎症对高血压刺激的反应。在目标3中，我们将使用外周血液T细胞 以确定T细胞ROCK2活性是否在 高血压。为了确定因果关系，我们将过继地将人类的T细胞转移到免疫缺陷小鼠身上 确定人类T细胞是否促进ROCK2患者的高血压和肾脏/血管炎症 人源化小鼠的依赖方式。我们的团队独一无二地准备进行这些翻译研究 可能会改变高血压及其相关并发症的免疫调节疗法。

PROJECT SUMMARY Hypertensive stimuli activate innate and adaptive immune cells which then act on target organs such as the kidney and vasculature causing tissue injury/inflammation and hypertensive end-organ damage. We and others have shown that depleting subsets of immune cells or individual cytokines in rodent models is indeed protective against these deleterious effects of hypertension. Yet there are currently no immunomodulatory therapies for hypertension, which affects nearly 50% of adults worldwide. A major reason for this is that immunosuppressive therapies would render a large percentage of people susceptible to infection and malignancy. A final common pathway of immune activation in experimental and human hypertension is the skewing of T cell subsets towards pro-inflammatory T helper 17 (Th17) cells, which secrete interleukin 17A (IL-17A) and IL-21, and away from anti- inflammatory regulatory T cells (Treg). There is emerging evidence that Rho-associated protein kinase 2 (ROCK2) acts as a molecular switch in T helper cells by promoting Th17 differentiation, via phosphorylation of STAT3 and IRF4, and inhibiting Treg differentiation. In this proposal, we will test the novel hypothesis that hypertension is associated with activation of a ROCK2/STAT3/IRF4 pathway in T cells that leads to increased Th17/Treg ratios and exacerbated hypertension and inflammatory damage. Furthermore, we propose that selectively inhibiting ROCK2 will restore a homeostatic T cell balance without causing global immunosuppression. A novel orally bioavailable selective ROCK2 inhibitor, KD025, has completed phase 2/3 clinical trials for IL-17A/IL-21 mediated autoimmune diseases with a favorable safety profile and no apparent increased risk of infection. We have novel preliminary data that KD025 significantly reduces BP and increases Treg/Th17 cell ratios in the kidney in response to experimental hypertension. In Aim 1 of this proposal, we will use inducible conditional ROCK2 deficient mice to determine cell-specific roles of ROCK2 in hypertension. To determine whether T cell ROCK2 deletion restores the Th17/Treg balance in hypertension with minimal effects on other cell types, we will use a novel single cell technique, called CITE-Seq, to phenotype circulating leukocytes as well as perform functional assays of immune function using a lipopolysaccharide (LPS) model of systemic inflammation. In Aim 2, we will determine the effect of KD025 on BP, Th17/Treg balance, end-organ damage, and LPS-induced inflammation in response to hypertensive stimuli. In Aim 3, we will use peripheral blood T cells from normotensive and hypertensive humans to determine whether T cell ROCK2 activity is increased in hypertension. To determine causality, we will adoptively transfer T cells from humans into immunodeficient mice to determine whether human T cells promote hypertension and renal/vascular inflammation in a ROCK2 dependent manner in humanized mice. Our team is uniquely poised to conduct these translational studies that will potentially transform immunomodulatory therapies for hypertension and its associated complications.