Immune Modulation in Hypertension

项目摘要 高血压刺激激活先天性和适应性免疫细胞，然后作用于靶器官，如 肾脏和脉管系统引起组织损伤/炎症和高血压终末器官损伤。我们和其他人 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， 对抗高血压的有害影响。然而，目前还没有免疫调节疗法， 高血压，影响全球近50%的成年人。一个主要原因是免疫抑制剂 治疗将使大部分人易受感染和恶性肿瘤的影响。最后一个共同点 实验性和人类高血压中的免疫激活途径是T细胞亚群向 促炎性T辅助17（Th 17）细胞，其分泌白细胞介素17 A（IL-17 A）和IL-21，并且远离抗- 炎性调节性T细胞（Treg）。有新的证据表明，Rho相关蛋白激酶2 （ROCK 2）作为辅助性T细胞中的分子开关，通过磷酸化 STAT 3和IRF 4，并抑制Treg分化。在这个建议中，我们将测试新的假设， 高血压与T细胞中ROCK 2/STAT 3/IRF 4通路的激活相关， Th 17/Treg比值与高血压及炎症损伤加重有关。此外，我们建议， 选择性抑制ROCK 2将恢复稳态T细胞平衡，而不会引起全身性炎症。 免疫抑制一种新型口服生物可利用的选择性ROCK 2抑制剂KD 025已完成2/3期研究 IL-17 A/IL-21介导的自身免疫性疾病的临床试验具有良好的安全性特征， 增加感染的风险。我们有新的初步数据，KD 025显着降低血压和增加 肾脏中Treg/Th 17细胞比率对实验性高血压的反应在本建议的目标1中，我们将 使用可诱导的条件性ROCK 2缺陷小鼠来确定ROCK 2在高血压中的细胞特异性作用。到 确定T细胞ROCK 2缺失是否以最小影响恢复高血压中的Th 17/Treg平衡 在其他类型的细胞上，我们将使用一种新的单细胞技术，称为CITE-Seq，来表型循环白细胞 以及使用脂多糖（LPS）模型进行免疫功能的功能测定， 炎症在目标2中，我们将确定KD 025对BP、Th 17/Treg平衡、终末器官损伤、 和LPS诱导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T细胞 从血压正常和高血压的人，以确定T细胞ROCK 2活性是否增加， 高血压为了确定因果关系，我们将过继性地将人类的T细胞转移到免疫缺陷小鼠中， 为了确定人T细胞是否促进ROCK 2中的高血压和肾/血管炎症， 在人源化小鼠中的依赖性方式。我们的团队是独一无二的准备进行这些转化研究， 将潜在地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.