(8) Mechanisms of autoimmune endocrine diseases in patients receiving checkpoint inhibitors

摘要 检查点抑制(CPI)疗法极大地改善了以前 被认为是难以对付的。由于这些药物具有预期的生物活性，从而能够激活T细胞 可导致肿瘤破坏的细胞，出现了新的自身免疫不良事件。这些措施的主要目标 不良反应一直是内分泌组织，包括甲状腺和胰腺中的β细胞。甲状腺炎是 频繁的自身免疫性糖尿病已经成为一种严重的不良事件，通常需要重症监护。 一些人发生这些自身免疫事件以及一些人受到保护的原因并不是 但这些信息可能会导致预防这些不良事件发生的方法。整体而言 目的了解自身免疫的分子和细胞免疫学基础。 糖尿病和甲状腺疾病，并测试是否可以通过特定的药物来预防它们 行为。我们想要检验的假设是，在患有内分泌疾病的个体中，CPI疗法 导致病理性T细胞、B细胞耐受性丧失和调节性T细胞功能障碍。我们的初步数据 已经确定了自体抗原反应性效应器T细胞和Tregs的表型差异 而不是发展成内分泌疾病。此外，我们对自身反应性B细胞的分析表明，CPI疗法 影响外周B细胞耐受检查点并导致自身反应性成熟频率增加 幼稚的B细胞。我们计划使用Seq-Well单细胞对前瞻性随访的患者进行分析 在治疗前到出现自身免疫性内分泌疾病时。在以下个体的子组中 是人类白细胞抗原A2(~40%)，我们将用细胞库和CyTOF研究甲状腺和糖尿病反应性CD8+T细胞 并确定在抗原反应细胞上发现的T细胞受体是否可以在 治疗以及在哪个亚群中。我们亦会决定消费物价指数会否影响 并具体说明CPI是否会导致Tregs产生病理性细胞因子。我们将使用 已建立的确定CPIs是否导致外周B细胞耐受失败的技术 找出B细胞和Treg细胞变化之间的关系。我们将收集有关自身反应性T细胞的数据 和B细胞谱系，我们将与原发肿瘤的临床反应相关。最后，我们对 这些不良事件背后的免疫学机制表明，它们可能以何种方式 预防，我们将在NOD小鼠抗PD-L1诱导的糖尿病小鼠模型中进行测试。我们将测试 B细胞耗尽或Tregs增强，无论是通过低剂量的IL-2还是通过输注糖尿病 抗原特异树突状细胞可以预防糖尿病的发生。因此，这些研究将阐明 这些严重的不良事件，确定谁是这些事件的最大风险的个人，并测试 可以使用不干扰CPIs的抗肿瘤效果的治疗方法来预防它们。

Summary Checkpoint inhibitor (CPI) therapy has greatly improved the treatment of cancers that had previously been considered intractable. As a result of the intended biologic activity of these drugs, which enable activation of T cells that can cause tumor destruction, new autoimmune adverse events have occurred. Major targets of these adverse events have been endocrine tissues including thyroid and beta cells in the pancreas. Thyroiditis is frequent and autoimmune diabetes has emerged as a serious adverse event often requiring intensive care. The reasons why some individuals develop these autoimmune events and why some are protected are not known but this information may lead to ways of preventing the occurrence of these adverse events. The overall objective if this proposal is to understand the molecular and cellular immunologic basis for autoimmune diabetes and thyroid disease and to test whether they can be prevented with agents that are specific in their actions. The hypothesis that we wish to test is that in individuals who develop endocrinopathies CPI therapy induces pathologic T cells, loss of B cell tolerance, and dysfunctional regulatory T cells. Our preliminary data has identified phenotypic differences in autoantigen reactive effector T cells and Tregs in those who do and do not develop endocrinopathies. In addition, our analysis of autoreactive B cells suggests that CPI therapy affects peripheral B cell tolerance checkpoints and results in an increased frequency of autoreactive mature naïve B cells. We plan to analyze, using Seq-Well single cells in patients who are followed prospectively from before treatment to when they present with autoimmune endocrinopathies. In the subgroup of individuals who are HLA-A2 (~40%) we will study thyroid and diabetes reactive CD8+ T cells with cellular libraries and CyTOF and determine whether the T cell receptors that are found on the antigen reactive cells can be detected prior to treatment and in which subpopulation. We will also determine whether CPIs affect the number and function of Tregs and address specifically whether the CPI causes the Tregs to produce pathologic cytokines. We will use established techniques to determine whether the CPIs induce a failure of peripheral B cell tolerance and identify the relationship between changes in B cells and Tregs. We will be collecting data on the autoreactive T and B cell repertoire that we will correlate with clinical responses to the primary tumors. Finally, our studies of the immunologic mechanisms that underlie these adverse events suggest ways in which they may be prevented, which we will test in a murine model of anti-PD-L1 induced diabetes in NOD mice. We will test whether B cell depletion or enhancement of Tregs, either by low doses of IL-2 or by infusion of diabetes antigen specific Tregs can prevent diabetes onset. These studies therefore, will elucidate the mechanisms of these serious adverse events, identify individuals who are at greatest risk for these events, and test whether therapies that do not interfere with the anti-tumor effects of the CPIs can be used to prevent them.