Role of Regulatory T-Lymphocytes in Chronic Heart Failure

调节性 T 淋巴细胞在慢性心力衰竭中的作用

基本信息

批准号：
8922490
负责人：
Sumanth D Prabhu
金额：
--
依托单位：
BIRMINGHAM VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8922490
关键词：
Ablation Adoptive Transfer Adult Age-Years American American Heart Association Anti-Inflammatory Agents Anti-inflammatory Antigen Presentation Antigen-Presenting Cells Antigens Apoptosis Biological Assay Blood Blood Circulation Blood capillaries Bone Marrow Boxing Bromodeoxyuridine C57BL/6 Mouse CD4 Positive T Lymphocytes CD8B1 gene Cardiac Cardiac Myoblasts Cells Characteristics Chronic Clinical Clinical Trials Congestive Heart Failure Coronary Data Dendritic Cells Diffuse Diphtheria Toxin Disease Disease Progression Drug or chemical Tissue Distribution EFRAC Economic Burden Elderly Endothelial Cells Exhibits Fibrosis Flow Cytometry Functional disorder Heart Heart Diseases Heart Hypertrophy Heart failure Home environment Hospitalization Human Immune Immunohistochemistry In Vitro Incidence Infarction Inflammation Inflammation Mediators Inflammatory Injury Interferons Knowledge Label Left Left Ventricular Remodeling Leukocytes Ligation Long-Term Effects Measures Mediastinal lymph node group Mediating Mediator of activation protein Mononuclear Mus Myocardial Ischemia Myocarditis Nuclear Patients Population Proteins Receptor Signaling Regulatory T-Lymphocyte Relative (related person)Role Signal Transduction Spleen Splenocyte Staging Stem cells Stroke System Systolic heart failure T-Cell Activation T-Lymphocyte T-Lymphocyte Subsets TNFRSF1A gene TNFRSF1B gene Testing Therapeutic Time Tissues Transcription Factor 3 Transgenic Mice Translating Tube Tumor Necrosis Factor Receptor Tumor Necrosis Factor-alpha Update Ventricular Veterans Woman Work angiogenesis base capillary cell type cellular targeting clinical practice cytokine density diphtheria toxin receptor immunoregulation improved in vivo indexing innovation lifetime risk lymph nodes macrophage men mortality neovascularization novel novel strategies novel therapeutic intervention outcome forecast p65 promoter public health relevance receptor reconstitution repository response statistics trafficking

项目摘要

DESCRIPTION (provided by applicant): No large-scale immunomodulatory clinical therapies currently exist for heart failure (HF). To date, the primary focus of therapeutic immunomodulation has been to neutralize inflammatory mediators such as tumor necrosis factor-α, an approach that unfortunately failed in clinical trial. Recently, we described profound remodeling of the global immune cell network and a heretofore-unappreciated adverse cardiosplenic axis in chronic ischemic HF that promotes immune cell-mediated tissue injury in the failing heart. This novel discovery suggests that directly targeting specific leukocyte populations that are altered in HF may represent a better approach to therapeutic immunomodulation. Regulatory T-cells (Tregs) are CD4+ T-lymphocytes that express the forkhead box protein-3 (Foxp3) transcription factor and suppress immune cell responses in a contactdependent, antigen nonspecific manner; their pathophysiological role in chronic HF is unknown. In our preliminary studies, we have unexpectedly discovered that in HF, Tregs robustly infiltrate the failing heart, and rather than being anti-inflammatory and thereby cardioprotective, paradoxically exhibit pro-inflammatory features and function prominently as disease mediators, imparting substantial anti-angiogenic, pro-fibrotic, and pro-inflammatory effects. Based on these data, we hypothesize that dysfunctional Tregs are essential for left ventricular (LV) remodeling, capillary rarefaction, inflammation, and disease progression in HF, and thereby are key cellular targets for immunomodulation. Three Aims will test this hypothesis. In Aim 1, we will delineate global Treg trafficking and alterations in Treg pro-inflammatory signaling in C57BL/6 mice with coronary ligation and HF, and in sham-operated controls. Tregs, and other T-cell subsets, and their inflammatory profiles will be defined in heart, blood, spleen, and mediastinal lymph nodes by flow cytometry and immunohistochemistry at serial time points after ligation, and correlated with LV and splenic remodeling and inflammation. BrdU labeling will index Treg proliferation in vivo. Moreover, using human HF biosample repositories, we will determine the abundance and distribution of Tregs in human failing hearts, and profile circulating Tregs (and other T-cells) in ambulatory patients with HF (with reduced ejection fraction) versus matched non-failing controls. In Aim 2, we will establish the role of Tregs in LV remodeling and neovascularization by selectively (and reversibly) depleting Tregs in Foxp3-DTR Tg mice with chronic HF, and then evaluating the effects on immune cell profiles, inflammation, circulating and bone marrow endothelial progenitor cells, in vivo angiogenesis, and LV remodeling. In parallel studies, the relative anti-angiogenic effects of splenic Tregs from sham mice, HF mice, and HF mice with ablation and subsequent reconstitution of Tregs will be compared using in vitro tube formation assays. In Aim 3, we will define whether dysfunctional Tregs are important mediators of the injurious cardiosplenic axis in chronic HF. CD45.2 mononuclear splenocytes from Foxp3-DTR Tg mice with either early or late HF, (corresponding to different stages of Treg dysfunction) will be adoptively transferred into CD45.1 naïve mice, with or without Treg ablation prior to transfer. The long-term effects of cell transfer on LV/splenicremodeling, inflammation, and T-cell/immune cell profiles will then be measured in recipient mice. In parallel, we will evaluate the ability of splenic Tregs from HF and sham mice to suppress activation of CD4+ and CD8+ splenic T-lymphocytes in vitro, and the role of TNFR signaling in HF Treg dysfunction via similar studies using splenic T-cells from TNFR1-/- and TNFR2-/- HF mice. By conclusively defining the role of Tregs in pathological cardiac remodeling, these studies will further our understanding of the cellular basis for inflammatory activation in chronic ischemic HF, and provide innovative perspectives as to the fundamental underpinnings of the recently discovered pro-inflammatory and tissue-injurious cardiosplenic axis. Moreover, by providing direct evidence for Tregs as anti-angiogenic and (paradoxically) pro-inflammatory mediators in HF, these studies will identify new targets for cell type-specific, rather than cytokine-specific, immunomodulation.

描述（由申请人提供）：目前没有大规模的免疫调节临床疗法（HF）。迄今为止，热免疫调节的主要重点是中和炎症介质（例如肿瘤坏死因子-α），这种方法不幸地在临床试验中失败了。最近，我们描述了全球免疫细胞网络的深刻重塑和慢性缺血性HF中迄今为止未贴心的不良心铂轴，从而促进了失败的心脏中免疫细胞介导的组织损伤。这个新颖的发现表明，直接靶向HF中改变的特定白细胞种群可能代表了一种更好的治疗性免疫调节方法。调节性T细胞（Tregs）是表达叉子盒蛋白3（FOXP3）转录因子的CD4+ T淋巴细胞，并以接触依赖性的，抗原非特异性方式抑制免疫球反应；他们在慢性HF中的病理生理作用尚不清楚。在我们的初步研究中，我们出乎意料地发现，在HF中，Tregs坚强地渗透了失败的心脏，而不是具有抗炎性，从而导致心脏保护性，矛盾地暴露于促疾病的促疾病和疾病介体的功能，作为疾病介体，启发性抗抗激素，亲纤维纤维纤维化，促纤维化效果和促进性效果。基于这些数据，我们假设Treg功能障碍对于左心室（LV）重塑，HF的毛细血管稀有，炎症和疾病进展至关重要，从而是必不可少的是免疫调节的关键细胞靶标。三个目标将检验这一假设。在AIM 1中，我们将描绘具有冠状动脉连接和HF的C57BL/6小鼠的Treg促炎信号传导的全球Treg贩运和变化，以及在假手术控制中。 TREG和其他T细胞亚群以及它们的炎症特征将通过流式细胞仪和连接后的串行时间点上的流式细胞仪和免疫组织化学定义，并与LV和脾脏重塑和炎症相关。 BRDU标签将在体内索引Treg的增殖。此外，使用人类HF生物样品存储库，我们将确定人类失败的心脏中Treg的抽象和分布，以及在HF患者中循环Treg（和其他T细胞）在HF（射血分数降低）中与匹配的非耐受性对照组的循环。在AIM 2中，我们将通过有选择地（和可逆地）用慢性HF在Foxp3-DTR TG小鼠中耗尽Treg，确定Tregs在LV重塑和新血管形成中的作用，然后评估对免疫细胞谱的影响，炎症，循环和骨骼莫罗型内皮细胞，vivo angieSiss和angi angi angiensepons，and angi angiensepons，and angi angiensephoseption and angi ongienseponsesepon and nivo angienogen和lv。重塑。在平行研究中，使用体外管形成测定法比较了Sham小鼠，HF小鼠和HF小鼠的脾脏Treg的相对抗血管生成作用。在AIM 3中，我们将定义功能失调的Treg是否是慢性HF中受伤的心铂轴的重要介体。 CD45.2来自FOXP3-DTR TG小鼠的单核脾细胞具有早期或晚期HF（对应于Treg功能障碍的不同阶段），将适当地转移到CD45.1幼稚的小鼠中，或者在转移前没有Treg消融。然后将在受体小鼠中测量细胞转移对LV/脾脏模块，炎症和T细胞/免疫细胞谱的长期影响。同时，我们将评估HF和假小鼠的脾脏Tregs的能力在体外抑制CD4+和CD8+脾脏T淋巴细胞的激活，以及TNFR信号在HF Treg功能障碍中使用类似的研究使用TNFR1 - / - 和TNFR2 - / - HF小鼠的脾T细胞的作用。通过最终定义TREG在病理心脏重塑中的作用，这些研究将进一步了解我们对慢性缺血性HF炎症激活的细胞基础的理解，并为最近发现的促炎和组织不良型卡片型轴的基本基础提供创新的观点。而且，通过提供直接 TREG作为抗血管生成和（自相矛盾的）促炎性介体的证据，这些研究将确定针对细胞类型特异性的，而不是细胞因子特异性，免疫调节的新靶标。