Manipulating DNA Damage-response Signaling for the Treatment of Type 1 Diabetes

操纵 DNA 损伤反应信号传导治疗 1 型糖尿病

基本信息

批准号：
10319938
负责人：
JONATHAN David KATZ
金额：
$ 44.68万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10319938
关键词：
Activated Lymphocyte Acute Address Allogenic Amputation Antigens Apoptotic Autoantigens Autoimmune Diseases Beta Cell Blindness Blood Glucose CD8-Positive T-Lymphocytes CD8B1 gene CHEK1 gene CHEK2 gene Cell Count Cell Cycle Checkpoint Cell Death Cells Chemosensitization Child Clinical Clinical Data Clinical Trials DNA Damage Data Diabetes Mellitus Disease Disease remission Drug Targeting Drug usage Equilibrium Eragrostis Exhibits Factor V Future Gamma-H2AX Genetic Transcription Genomics Harvest Hour Human Hyperglycemia Immune Immune System Diseases Immune Targeting Immune mediated destruction Immune response Immune system Immunity Immunotherapy Impairment Inbred NOD Mice Incidence Individual Infection Insulin Insulin-Dependent Diabetes Mellitus Islets of Langerhans Islets of Langerhans Transplantation Kidney Diseases Lead Life Lymphocyte Lymphocyte Biology MDM2 gene Malignant Neoplasms Memory Molecular Mechanisms of Action Molecular Target Monitor Mus Neuropathy Pancreas Pathogenicity Pathologic Patients Pharmaceutical Preparations Phosphorylation Population Property Publishing Regulatory T-Lymphocyte Role Sampling Signal Pathway Signal Transduction Spleen Stress Stroke Structure of beta Cell of islet T memory cell T-Lymphocyte TP53 gene Testing Therapeutic Tissues Toxic effect Transplantation Tolerance Treatment Cost Vascular Diseases autoreactivity base blood glucose regulation diabetogenic effector T cell efficacy testing experimental study fighting genetic risk factor glucose monitor immunoregulation in vivo insight insulin dependent diabetes mellitus onset islet lymph nodes non-diabetic novel novel therapeutics pre-clinical preservation prevent response therapeutic target vaccine-induced immunity young adult

项目摘要

Abstract Type 1 diabetes (T1D) is a common autoimmune disease in children and young adults. T1D presents as acute onset hyperglycemia resulting from the immune-mediated destruction of insulin-producing pancreatic beta cells. The central pathogenic driver of T1D is the beta cell antigen-specific (ag.-sp.) T cell. There is no durable cure for T1D; the sole and costly treatment for T1D remains daily insulin replacement. Even with vigilant glucose monitoring and control, T1D patients still suffer a host of life-threatening sequalae including macro- and micro- vasculopathies, neuropathy, nephropathy, amputations, stroke, and blindness. While progress has been made in (i) producing and delivering insulin, (ii) monitoring blood glucose, (iii) identifying autoantigens, (iv) defining genetic risk factors, (v) understanding underlying immune dysfunction, and (vi) producing and harvesting pancreatic islet cells for transplant, the most intractable barrier remains our inability to remove or control islet ag.-sp. T cells, without which the promise of preventing/curing T1D will likely fail. To surmount this critical barrier, we devised the means to eliminate diabetogenic T cells from the adaptive immune repertoire. In fact, when applied to non-obese diabetic (NOD) mice with spontaneous new-onset T1D, we observe (i) a striking prolongation of the remission or “honeymoon” period, (ii) a significant reduction in beta cell-specific CD4+ and CD8+ T cells, (iii) a significant preservation of beta cells, and (iv) a highly significant reduction (78%) in the number of NOD mice that transit to overt diabetes. The premise: As T cells toggle between distinct states – naïve, activated effector, quiescent and activated memory – they exhibit ineluctable properties that we can precisely target. This is particularly true of activated effector CD4+ and CD8+ T cells (Teff). Unlike their counterparts, Teff cells divide rapidly – at a rate of once every 5-6 hours in vivo – and exhibit an intrinsic DNA damage response (DDR) that places them on the edge of apoptotic cell death. We hypothesize (i) that this unique aspect of lymphocyte biology lead to genomic stress in acutely activated lymphocytes and (ii) that manipulation of DDR signaling pathways allows for selective therapeutic targeting of pathological T cells. Consistent with these hypotheses, we find that both mouse and human Teff cells display a pronounced DDR, as evidenced by DNA damage, phospho-ser139 H2AX (γH2AX), and phosphorylation of ATM, CHK2, and p53. Moreover, we find that novel drugs that potentiate p53 (via inhibition of MDM2) or impair cell cycle checkpoints (via inhibition of CHK1/2 or WEE1) lead to the selective elimination of pathological Teff cells in vivo when given during a prescribed therapeutic window. In combination of these compounds – which we termed “p53 potentiation with checkpoint abrogation” (PPCA) – display clear therapeutic benefit, targeting pathological T cells but does not naive, regulatory, or quiescent memory T-cell pools, and has a modest nonimmune toxicity profile. These results, recently published, (PNAS 2017, PMC5474825) suggest a novel and tractable clinical strategy for a highly selective form of immune therapy that is (i) specific for both CD4+ and CD8+ auto-reactive Teff cells, (ii) minimally or non-genotoxic, and (iii) markedly better tolerated than current approaches. Importantly, this approach does not alter tissue-resident Treg cell numbers; in fact, our data suggest that PPCA resets the regulatory balance in favor of Treg control of anti-beta cell immunity. Based on our preliminary and published data, we propose three inter-related hypotheses: (i) that PPCA has a distinct mechanism of action that eliminates Teff cells while sparing Treg cells, thereby re- establishing a localized regulatory balance; (ii) that PPCA can target the control of both auto- and allogeneic T cells, thereby allowing for sustained transplantation tolerance to islets, and (iii) that PPCA can preferentially target islet ag.-sp. activated human T cells in individuals with T1D while sparing the memory compartment.

抽象的 1型糖尿病（T1D）是儿童和年轻人常见的自身免疫性疾病。 T1D礼物由于免疫介导的胰岛素产生胰腺破坏导致急性发作高血糖症 β细胞。 T1D的中心致病驱动器是β细胞抗原特异性（AG.-SP.）T细胞。没有耐用治疗T1D； T1D的唯一且昂贵的治疗仍然每天替代胰岛素。即使有警惕的葡萄糖监测和控制，T1D患者仍然遭受危及生命的频谱血管病，神经病，肾病，截肢，中风和失明。虽然进展已取得（i）产生和输送胰岛素，（ii）监测血糖，（iii）识别自身抗原，（iv）定义遗传危险因素，（v）理解潜在的免疫功能障碍，以及（vi）产生和收获用于移植的胰岛细胞，最棘手的屏障仍然我们无法移除或控制胰岛 AG.-SP. T细胞没有预防/固化T1D的承诺可能会失败。为了克服这一关键障碍，我们设计了消除从适应性免疫曲目。实际上，当应用于非肥胖的糖尿病（点头）小鼠时新发行的T1D，我们观察（i）缓解或“蜜月”时期的惊人延长，（ii）显着 β细胞特异性CD4+和CD8+ T细胞的降低，（iii）对β细胞的显着保存，以及（iv）高度转向明显的糖尿病的NOD小鼠的数量显着降低（78％）。前提：当T细胞在不同状态之间切换时 - 幼稚，激活效应子，静止和激活记忆 - 它们暴露了我们可以精确针对的不可避免的属性。激活尤其如此效应子CD4+和CD8+ T细胞（TEFF）。与他们的对应物不同，Teff细胞迅速分裂 - 每次一次 5-6小时在体内 - 表现出固有的DNA损伤响应（DDR），将它们置于凋亡细胞死亡。我们假设（i）（i）淋巴细胞生物学的这一独特方面导致基因组应激在急性激活的淋巴细胞和（ii）操纵DDR信号通路可以选择性的病理T细胞的治疗靶向。与这些假设一致，我们发现鼠标和人类的Teff细胞显示出明显的DDR，如DNA损伤所证明的，磷酸ser139 H2AX（γH2AX）， ATM，CHK2和P53的磷酸化。此外，我们发现具有潜在p53的新型药物（通过抑制MDM2）或损害细胞周期检查点（通过抑制CHK1/2或WEE1）导致选择性在规定的治疗窗口中给予病理Teff细胞体内的病理Teff细胞。结合在这些化合物中 - 我们称其为“ p53用检查点”（ppca）显示清晰治疗益处，靶向病理T细胞，但不天真，调节或静态记忆T细胞T细胞T细胞池，具有适度的非免疫毒性特征。这些结果，最近发布的（PNAS 2017， PMC5474825）提出了一种新型且可进行的临床策略，以一种高度选择性的免疫疗法形式是（i）针对CD4+和CD8+自动反应性TEFF细胞（ii）（ii）最小或非生物毒性的特异性，以及（iii）比当前方法明显好。重要的是，这种方法不会改变组织居民 Treg单元格数；实际上，我们的数据表明PPCA重置了监管平衡，以控制Treg的控制抗β细胞免疫。根据我们的初步和已发布的数据，我们提出了三个相互关联的假设：（i） PPCA具有独特的作用机理，可以消除Teff细胞，同时保留Treg细胞，从而重复使用建立局部监管平衡；（ii）PPCA可以针对自身和同种异体T的控制细胞，从而允许对胰岛进行持续的移植耐受性，并且（iii）PPCA可以优先使用PPCA 目标胰岛AG.-SP.在保留记忆室的同时，激活了具有T1D个体的人T细胞。