Mechanism of  PD1 on cardiac inflammation resolution during heart failure development

心血管炎症促进心力衰竭(HF)的发展。然而，心脏的作用机制 心力衰竭发展过程中的炎症消退仍然知之甚少。程序性细胞死亡蛋白1(PD1) 是一种蛋白质，通过抑制最初的T细胞诱导和通过 维持T细胞耐受性。PD1阻断抗体用于癌症治疗，但治疗也导致 对某些患者的心脏毒性有影响。我们发现PD1KO或PD1封闭抗体显著加剧 横断性主动脉缩窄(TAC)引起的心脏炎症、心衰和死亡，表明PD1发挥了更多的 在压力条件下发挥重要作用。为了了解PD1抑制心脏炎症的机制，我们 假手术和TAC对野生型和PD1KO小鼠心脏免疫细胞和血管内皮细胞的影响 用单细胞CITE-SEQ结合条形码抗体进行膜蛋白标记。使用单电池 Cite-Seq，我们还研究了HF小鼠和假手术小鼠的肺免疫细胞。生物信息学分析已经 提供了这些细胞的大量信息-显示了细胞团的戏剧性变化，丰富了途径 先天免疫反应和获得性免疫反应及免疫细胞亚群代谢途径的变化 心衰小鼠，或TAC后PD1KO。GDT细胞(T细胞的一个亚群)可分为IL-17(GdT17)或IFNG 制片人。肺免疫细胞CITE-SEQ结果显示，HF可引起多种T细胞和T细胞的显著变化 巨噬细胞簇，在Th17和gdT17细胞中PD1的表达急剧增加，提示PD1在 在抑制Th17和gdT17细胞以及HF进展中的作用。心脏免疫细胞中的CITE-SEQ显示 PD1KO小鼠TAC后CD8T细胞和GDT细胞的浸润增加，这些细胞 提示CD8T细胞、GDT细胞和IFNG可能参与了心脏疾病的加重。 PD1KO小鼠的炎症反应。基于这些令人兴奋的发现，我们假设TAC诱导的心脏和 PD1至少通过保守和独特的途径调节肺部炎症的消退 部分受CD8T细胞、GDT细胞和Th17细胞产生的IFNG和IL17控制。为了增强 我们对PD1在心脏炎症和心力衰竭中的作用的研究具有创新的严谨性，我们还将研究CD8 细胞特异性PD1KO小鼠。AIM-1。验证IFNG和CD8 T细胞参与急性加重期的假设 TAC后PD1KO的心脏炎症、细胞因子风暴和心力衰竭。此外，我们将确定PD1是否 CD8T细胞中的KO足以加重TAC诱导的心脏炎症和心衰。AIM-2。确定 IL-17和GDT细胞在TAC诱导的心脏炎症和心衰中的作用及其机制 PD1抑制后。本申请高度响应特别关注通知NOT-ES-20-018作为 拟议的研究将促进我们对PD1和T细胞在心脏和肺中的作用机制的理解 炎症消退与心肺免疫细胞团保守而独特的变化 心衰的发展和进展。

Cardiovascular inflammation promotes Heart failure (HF) development. However, mechanism of cardiac inflammation resolution during HF development is still poorly understood. Programmed cell death protein 1 (PD1) is a protein that keeps the body’s immune responses in check, both by inhibiting initial T cell induction and by maintaining T cell tolerance. PD1 blocking antibodies are used in cancer treatment, but the treatment also leads to cardiac toxicity in some patients. We found that PD1 KO or PD1 blocking antibodies dramatically exacerbated transverse aortic constriction (TAC)-induced cardiac inflammation, HF, and death, indicating PD1 exerts a more important role under stress conditions. To understand mechanisms of PD1 inhibition in cardiac inflammation, we studied cardiac immune cells and vascular endothelial cells from wild type and PD1 KO mice after sham or TAC by using single-cell CITE-seq together with barcoded antibodies for membrane protein labeling. Using single-cell CITE-seq, we also studied lung immune cells from HF mice and sham mice. Bioinformatics analyses have provided enormously information of these cells – showing dramatic alterations of cell clusters, enriched pathways of innate & adaptive immune responses, and changes of metabolic pathways in various immune cell subsets in HF mice, or in PD1 KO after TAC. gdT cells (a subset of T cells) can be divided into either IL-17 (gdT17) or IFNg producers. CITE-seq of lung immune cells showed that HF caused dramatic changes of various T cell and macrophage clusters, a dramatic increase of PD1 in Th17 and gdT17 cells, suggesting PD1 exerts an important role in suppressing Th17, and gdT17 cells as well as HF progression. CITE-seq in cardiac immune cells showed that infiltration of CD8+ T cells and gdT cells increased in PD1 KO mice after TAC, and these infiltrated cells are IFNg+ cells, indicating that CD8+ T cells, gdT cells, and IFNg may contribute to the exacerbated cardiac inflammation in PD1 KO mice. Based on these exciting findings, we hypothesize that TAC-induced cardiac and pulmonary inflammation resolution is regulated by PD1 through both conserved and unique pathways at least partially controlled by IFNg and IL17 produced by CD8+ T cells, gdT cells, and Th17, respectively. To enhance the innovative rigor of our investigation of the role of PD1 in cardiac inflammation and HF, we will also study CD8 cell specific PD1 KO mice. Aim-1. Test the hypothesis that IFNg and CD8+ T cells contribute to the exacerbated cardiac inflammation, cytokine storm, and HF in PD1 KO after TAC. In additon, we will determine whether PD1 KO in CD8+ T cells is sufficient to exacerbate TAC-induced cardiac inflammation and HF. Aim-2. Determine the roles and underlying mechanisms of IL17 and gdT cells in promoting TAC-induced cardiac inflammation and HF after PD1 inhibition. This application is highly responsive to the Notice of Special Interest NOT-ES-20-018 as the proposed studies will advance our understanding of the mechanisms of PD1 and T cells in cardiac and lung inflammation resolution, and the conserved & unique changes in cardiac and lung immune cell clusters during HF development and progression.