Lipid Antigen Presentation as a Driver of T2D Inflammation

项目总结 大量的研究工作发现慢性炎症在胰岛素的发展中起着关键作用 抵抗，2型糖尿病(T2D)的一个标志。即使有了这一发现，也没有成功的抗炎药物 治疗学已经得到发展，但研究停滞不前，重点放在脂肪组织的特征上 T2D小鼠模型的炎症反应。重要的是，T2D中慢性炎症的发起者仍然存在 未知。2型糖尿病表现为针对多种自身蛋白的自身抗体，但蛋白 抗体反应性筛查尚未发现T2D炎症的起始者。值得注意的是，并不是所有 T2D中的自身抗原是蛋白质。我们已经在T2D中发现了完全 被忽视，可能是因为呈现脂类抗原的蛋白质，第一组CD1分子没有表达 在老鼠身上。这种表达的缺乏给我们在免疫学中对脂类抗原的理解留下了一个巨大的漏洞 动态平衡与疾病，尤其是T2D。为了解决这个问题，我们将使用转基因小鼠来表达 人类CD1分子和人类免疫反应的体外模型以了解脂质介导的免疫 在T2D中的函数。这项提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。这一建议验证了最重要的假设，即脂类抗原呈递是一种关键机制，通过 T2D的慢性炎症是由哪种因素引起的。这是一项创新和新颖的脂类抗原研究 在T2D中展示结合了使用人类血液样本、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，再加上计算方法 允许采用系统生物学的方法来发现抗原和检验假说。在研究领域1，我们将 评估T细胞反应的脂类抗原依赖性激活。我们将使用人源化的老鼠和人类的血液 以询问这些脂质抗原对T细胞功能的影响(CD1四聚体负载)和 体内(高脂饮食)。在研究领域2，我们将以脂类抗原递呈为靶点来预防和治疗T2D。这个 针对第1组CD1分子和脂类自身抗原的中和抗体抑制能力 人类免疫细胞分泌的促炎细胞因子及其在转基因防治疾病中的作用 将对老鼠进行评估。在研究领域3.在T2D中发现脂质自身抗原将使用一种新的 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 脂类抗原提呈引起的自身免疫性疾病和彻底改变治疗策略 疾病管理和治疗发展。这项提案的结果将使 确定可作为疾病治疗靶点的特定炎症途径。

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.