Intracellular Targeting of Proinflammatory Pathways to Ameliorate Type 1 Diabetes

描述（由申请人提供）：1型糖尿病（T1D）是一种慢性自身免疫性疾病，具有长期发病率和死亡率的高风险。虽然胰岛素可以为受影响的儿童和成人提供相对正常的生活，但该治疗存在癫痫发作，昏迷和死亡的风险，即使是依从的患者也不总是能避免并发症，如失明，肾衰竭和截肢。我们目前对这种疾病的理解描绘了一幅复杂的画面，几乎每个免疫细胞都参与其中。这些相互作用包括来自T细胞、B淋巴细胞、树突细胞和巨噬细胞的必要贡献。这些细胞的激活导致组织炎症状态和促炎细胞因子和其他有害炎症产物对产生胰岛素的胰岛β细胞的最终破坏。为了激活与炎症状态相关的转录程序，这些有害信号必须从细胞表面转导到细胞核。这一过程需要促炎转录因子的核输入，并依赖于核转运蛋白/输入蛋白复合物的作用。我们已经开发了一种细胞穿透抑制剂的核输入抑制加速糖尿病和胰岛炎一年的NOD小鼠，最广泛使用的人类疾病模型。在这项提案中，我们将测试中心假设，即针对炎症反应所需的两个关键控制点将改善并可能逆转自身免疫性糖尿病。在目标1中，我们将确定核输入抑制剂在预防或逆转自发性糖尿病中的功效，并阐明其作用机制。炎症的第二个关键步骤是激活先天免疫的支柱，Toll样受体（TLR）。这些受体与糖尿病发病机制有关，它们的激活可破坏免疫耐受。通过这些受体的信号传导依赖于两个关键的衔接蛋白，MyD88和TRIP。在具体目标2中，我们将开发这些途径的细胞穿透抑制剂，并测试它们在抑制或逆转1型糖尿病方面的功效。总的来说，我们将使用创新的细胞穿透肽和蛋白质靶向促炎信号的近端（衔接蛋白激活）和远端（核转位）控制点。靶向这些关键的细胞内信号传导事件代表了改善1型糖尿病的新策略。公共卫生相关性：1型糖尿病是一种慢性疾病，始于儿童时期，困扰着200多万美国人。那些受影响的人面临终身注射，而不确定他们将免受疾病并发症的影响。我们将测试T1D发展的新途径，并应用创新方法来靶向这些途径，以揭示预防和逆转这种毁灭性疾病的新策略。

DESCRIPTION (provided by applicant): Type 1 diabetes (T1D) is a chronic autoimmune disease with high risk for long-term morbidity and mortality. Although insulin can provide a relatively normal life for the affected children and adults, the therapy poses risk for seizures, coma, and death and even compliant patients are not always protected from complications such as blindness, renal failure, and amputation. Our current understanding of the disease paints a complex picture in which nearly every immune cell is involved. These interactions include requisite contributions from T cells, B lymphocytes, dendritic cells, and macrophages. The activation of these cells results in a state of tissue inflammation and ultimate destruction of insulin-producing islet beta cells by proinflammatory cytokines and other noxious products of inflammation. To activate the transcriptional program associated with the inflammatory state, these deleterious signals must be transduced from the cell surface to the nucleus. This process requires the nuclear import of proinflammatory transcription factors and depends on the action of karyopherin/importin complexes. We have developed a cell-penetrating inhibitor of nuclear import which suppresses accelerated diabetes and insulitis for one year in NOD mice, the most widely used model of human disease. In this proposal, we will test the central hypothesis that targeting two key control points required for the inflammatory response will ameliorate and possibly reverse autoimmune diabetes. In aim 1, we will determine the efficacy of a nuclear import inhibitor in preventing or reversing spontaneous diabetes and will elucidate its mechanism of action. A second key step in inflammation is the activation of the mainstays of innate immunity, toll-like receptors (TLRs). These receptors have been implicated in diabetes pathogenesis and their activation can break immune tolerance. Signaling through these receptors depends on two key adaptor proteins, MyD88 and TRIP. In specific aim 2, we will develop cell-penetrating inhibitors of these pathways and test their efficacy in suppressing or reversing Type 1 diabetes. Overall, we will target both the proximal (adaptor protein activation) and distal (nuclear translocation) control points of proinflammatory signaling using innovative cell-penetrating peptides and proteins. Targeting these crucial intracellular signaling events represents a new strategy to ameliorate Type 1 diabetes. PUBLIC HEALTH RELEVANCE: Type 1 diabetes is a chronic illness that begins in childhood and afflicts more than 2 million Americans. Those affected face a life-time of injections without certainty that they will be protected from disease complications. We will test new pathways in T1D development and apply innovative approaches to target these pathways to reveal new strategies for prevention and reversal of this devastating illness.