Sex Differences in lipid antigen presentation, impact of lipid antigen presentation on peripheral lipid metabolism

项目摘要 大量的研究工作已经发现了慢性炎症在胰岛素产生中的关键作用 2型糖尿病（T2D）的症状有哪些？即使有了这个发现，也没有成功的抗炎药 治疗方法已经被开发出来，研究已经停滞，重点是表征脂肪组织 T2D小鼠模型中的炎症。重要的是，T2D中慢性炎症的起始物仍然是 未知2型糖尿病表现为针对多种自身蛋白质的自身抗体，但蛋白质 抗体反应性的筛选尚未发现T2D炎症的引发剂。值得注意的是， T2D中的自身抗原是蛋白质。我们已经鉴定出T2D中的脂质自身抗原， 被忽视，可能是由于呈递脂质抗原的蛋白质，即第1组CD1分子，未被表达 对小鼠这种表达的缺乏在我们对免疫学中脂质抗原的理解中留下了一个巨大的漏洞。 体内平衡和疾病，尤其是T2D。为了解决这个问题，我们将使用转基因小鼠， 人类CD1分子和人类免疫应答的体外模型，以了解脂质介导的免疫 T2D功能这项建议的长期目标是建立一个新的科学方向， 了解脂质在促炎反应、T2D和自身免疫中的作用的长期目的 疾病该提案测试了总体假设，即脂质抗原呈递是一个关键机制， T2D中的慢性炎症开始。这项创新和新颖的研究脂质抗原 T2D中的呈递结合使用人血液样品、转基因和人源化小鼠、单细胞RNA 测序，多参数细胞因子分析和流式细胞术，加上计算方法， 允许系统生物学方法来发现抗原和检验假设。在研究领域1，我们将 评估T细胞应答的脂质抗原依赖性活化。我们将使用人源化的老鼠和人血 样品以询问这些脂质抗原对体外T细胞功能的影响（CD1四聚体加载）， 体内（高脂肪饮食）。在研究领域2中，我们将靶向脂质抗原呈递以预防和治疗T2D。的 靶向第1组CD1分子和脂质自身抗原的中和抗体抑制 从人免疫细胞分泌的促炎细胞因子以及预防和治疗转基因动物中的疾病 将对小鼠进行评估。在研究领域3。T2D中的脂质自身抗原发现将使用一种新的 用于抗体分析的3D打印脂质微阵列。该提案有可能将T2D重新定义为一种 脂质抗原呈递驱动的自身免疫性疾病，并完全改变 疾病管理和治疗开发。该提案的结果将允许 识别可用于疾病治疗的特定炎症途径。

PROJECT SUMMARY Substantial research effort has uncovered a critical role for chronic inflammation in the development of insulin resistance, a hallmark of Type 2 Diabetes (T2D). Even with this discovery, no successful anti-inflammatory therapeutics have been developed, and research has stagnated with a focus on characterizing adipose tissue inflammation in mouse models of T2D. Importantly, the initiator of chronic inflammation in T2D remains unknown. Type 2 Diabetes presents with autoantibodies against a wide array of self-proteins, but protein screens for antibody reactivity have not discovered initiators of T2D inflammation. Remarkably, not all autoantigens in T2D are protein. We have identified lipid autoantigens in T2D that have been completely overlooked, likely due to the proteins which present lipid antigen, Group 1 CD1 molecules, not being expressed in mice. This lack of expression has left a gaping hole in our understanding of lipid antigens in immunological homeostasis and disease, especially T2D. To remedy this issue, we will use transgenic mice which express human CD1 molecules and in vitro models of human immune responses to understand lipid-mediated immune function in T2D. The long-term goal of this proposal is to establish a new scientific direction with the broad long-term purpose of understanding the role of lipids in pro-inflammatory responses, T2D, and autoimmune diseases. This proposal tests the overarching hypothesis that lipid antigen presentation is a key mechanism by which chronic inflammation in T2D is initiated. This innovative and novel investigation of lipid antigen presentation in T2D combines use of human blood samples, transgenic and humanized mice, single cell RNA sequencing, multiparameter cytokine analysis and flow cytometry, coupled with computational methods that allow for a systems biology approach to antigen discovery and hypotheses testing. In research Area 1, we will assess lipid antigen dependent activation of T cell responses. We will use humanized mice and human blood samples to interrogate the impact of these lipid antigens on T cell function in vitro (CD1 tetramer loading) and in vivo (high fat diet). In research Area 2, we will target lipid antigen presentation to prevent and treat T2D. The capacity of neutralizing antibodies targeted against group 1 CD1 molecules and lipid autoantigens to inhibit pro-inflammatory cytokines secreted from human immune cells and to prevent and treat disease in transgenic mice will be assessed. In research Area 3. Lipid autoantigen discovery in T2D will be performed with a novel 3D printed lipid microarray for antibody profiling. This proposal has the potential to redefine T2D as an autoimmune disease driven by lipid antigen presentation and completely change the strategies for management of disease and therapeutic development. Outcomes of this proposal will allow for the identification of specific inflammatory pathways that can be targeted for disease treatment.