Influence of T cell genotype/phenotype in atherosclerotic cardiovascular disease

项目总结 动脉粥样硬化性心血管疾病(ASCVD)是一种发病机制不明的炎症性疾病 仍然是全世界发病率和死亡率的主要原因。免疫反应起着核心作用。 在这种慢性疾病的演变过程中，CD4CD25Foxp3调节性T细胞(Tregs)发挥着重要的作用 调节炎症。我们已经显示了CD39的表达水平，一种免疫抑制剂 人类特异性单核苷酸多态与Tregs上的胞外核苷酸酶 (SNPs)。尽管CD39的特定SNP与克罗恩病有关，但还没有研究证实 Treg CD39催化活性的变化和腺苷清除细胞外ATP的变化 ASCVD的产生、影响、表现。同样，自然产生的反义ENTPD1-AS1已经被 研究表明，CD39在克罗恩病患者树中的表达减少。特异性自身对ENTPD1-AS1的抑制作用 传递Fana CD39反义(Fana-CD39-AS)寡核苷酸被证明增加CD39的表达， 提供了一种调节Treg表型的治疗选择。我们的总体实验目标是阐明 基因调控对ASCVD中Treg CD39表达的影响中心假设是基因突变 导致Treg CD39活性降低和嘌呤能反应改变导致ASCVD 充分消炎。我们将通过进行功能基因组实验来验证这一假设， 除了使用新的小鼠实验模型系统，并开发临床研究，检查分离的 细胞和病人的生物标本。该提案包括两个目标。在SA1中，我们的调查小组将确定 验证的实验系统中CD39活性的Treg表达如何影响动脉粥样硬化。这将是 使用CD39 Treg特异性条件性基因敲除和基因敲除小鼠模型，其多个读数为 T细胞和髓系激活反应，以及过继转移研究，我们将检查在SA2中， CD39在人Tregs上的表达，并检测CD39对免疫功能和免疫功能的影响 ASCVD临床研究中的炎症反应。我们已经组建了一个协作团队，与临床和 在ASCVD、免疫学、血管生物学、遗传学和生物统计学方面的实验专业知识。已完成的 提议的AIMS将加深对Tregs的作用的理解，特别是CD39和改变的表达 在ASCVD中，这种最重要和最显著的疾病是嘌呤能反应。转化为临床实践将会 通过识别重要的生物标记物和新的靶点，包括CD39和相关途径，来促进 腺苷能信号转导，用于ASCVD的治疗干预。

PROJECT SUMMARY Atherosclerotic cardiovascular disease (ASCVD) is an inflammatory disease of unclear pathogenesis that remains the leading cause of morbidity and mortality throughout the world. Immune responses play a central role in the evolution of this chronic disease, and CD4+CD25+Foxp3+ regulatory T cells (Tregs) exhibit critical functions to regulate inflammation. We have shown the level of expression of CD39, an immunosuppressive ectonucleotidase, on Tregs to be genetically associated in humans with specific single nucleotide polymorphisms (SNPs). Although select SNPs for CD39 have been linked to Crohn disease, no studies have examined whether alterations in Treg CD39 catalytic activity, and changes in extracellular ATP scavenging with adenosine generation, impact manifestations of ASCVD. Similarly, a naturally occurring antisense ENTPD1-AS1 has been shown to decrease CD39 expression in Crohn’s patient Tregs. Inhibition of ENTPD1-AS1 with a specific self- delivering FANA CD39 antisense (FANA-CD39-AS) oligonucleotide were shown to increase CD39 expression, providing a therapeutic option to modulate Treg phenotype. Our overall experimental goal is to elucidate the impact of genetic regulation of Treg CD39 expression in ASCVD. The central hypothesis is that genetic mutations resulting in decreased Treg CD39 activity and altered purinergic responses drive ASCVD due to the inability to adequately resolve inflammation. We will test this hypothesis by conducting functional genomic experiments, in addition to using novel murine experimental model systems, and develop clinical studies, examining isolated cells and patient biospecimens. The proposal consists of two Aims. In SA1, our investigative team will determine how Treg expression of CD39 activity impacts atherosclerosis in a validated experimental system. This will be done using CD39 Treg-specific conditional knock-down and knockout murine models with multiple readouts of T cell and myeloid activation responses, and adoptive transfer studies, In SA2, we will examine the regulation of CD39 expression on human Tregs and examine the impact of CD39 modulation on immune function and inflammation in clinical studies of ASCVD. We have assembled a collaborative team with clinical and experimental expertise in ASCVD, immunology, vascular biology, genetics, and biostatistics. Completion of the proposed aims will develop understanding of the role of Tregs, and specifically expression of CD39 and altered purinergic responses, in ASCVD, this most important and significant disease. Translation to clinical practice will be facilitated by identification of important biomarkers and novel targets, inclusive of CD39 and related pathways of adenosinergic signaling, for therapeutic intervention in ASCVD.