Novel therapeutic intervention of early-stage T1D

早期 T1D 的新型治疗干预

基本信息

批准号：
10698534
负责人：
XIAN CHEN
金额：
$ 30.07万
依托单位：
TRANSCHROMIX, LLC.
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-22 至 2024-06-21
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10698534
关键词：
Affect Autoimmunity Automobile Driving B-Lymphocytes Beta Cell Binding Biotinylation Cells Chromatin Clinical Data Diabetes Mellitus Diabetes prevention Disease Disease Pathway Disease Progression Early Intervention Epidemiology Event Funding Generations Genes Genetic Transcription Heterogeneity Human Inbred NOD Mice Infiltration Insulin Insulin-Dependent Diabetes Mellitus Islets of Langerhans Lymphocyte Measures Mediating Modeling Mus Organ Pancreas Pancreatic Diseases Pathogenesis Pathogenicity Pathology Pathway interactions Patients Peripheral Blood Mononuclear Cell Phase Protac Protein Synthesis Inhibition Proteins Proteome Proteomics Replacement Therapy Sampling Specificity Splenocyte Surface T cell infiltration T-Cell Immunologic Specificity T-Cell Proliferation Technology Testing Therapeutic Time Toxic effect Translational Regulation Translational Repression Translations Validation autoreactivity cell type chemokine chemoproteomics chronic autoimmune disease companion diagnostics cytokine cytotoxic diabetes mellitus therapy diabetes pathogenesis diagnostic assay drug action effector T cell energy efficiency genetic regulatory protein glucose metabolism histone methyltransferase immune cell infiltrate immunomodulatory therapies in vivo inhibitor innovation insulin secretion insulitis islet medical schools novel novel strategies novel therapeutic intervention patient stratification pre-clinical prevent professor programs protein expression reduce symptoms response small molecule targeted treatment therapeutically effective translatome virulence gene

Affect Autoimmunity Automobile Driving B-Lymphocytes Beta Cell Binding Biotinylation Cells Chromatin Clinical Data Diabetes Mellitus Diabetes prevention Disease Disease Pathway Disease Progression Early Intervention Epidemiology Event Funding Generations Genes Genetic Transcription Heterogeneity Human Inbred NOD Mice Infiltration Insulin Insulin-Dependent Diabetes Mellitus Islets of Langerhans Lymphocyte Measures Mediating Modeling Mus Organ Pancreas Pancreatic Diseases Pathogenesis Pathogenicity Pathology Pathway interactions Patients Peripheral Blood Mononuclear Cell Phase Protac Protein Synthesis Inhibition Proteins Proteome Proteomics Replacement Therapy Sampling Specificity Splenocyte Surface T cell infiltration T-Cell Immunologic Specificity T-Cell Proliferation Technology Testing Therapeutic Time Toxic effect Translational Regulation Translational Repression Translations Validation autoreactivity cell type chemokine chemoproteomics chronic autoimmune disease companion diagnostics cytokine cytotoxic diabetes mellitus therapy diabetes pathogenesis diagnostic assay drug action effector T cell energy efficiency genetic regulatory protein glucose metabolism histone methyltransferase immune cell infiltrate immunomodulatory therapies in vivo inhibitor innovation insulin secretion insulitis islet medical schools novel novel strategies novel therapeutic intervention patient stratification pre-clinical prevent professor programs protein expression reduce symptoms response small molecule targeted treatment therapeutically effective translatome virulence gene

展开

项目摘要

Abstract Type 1 diabetes (T1D) is a chronic autoimmune disease in which insulin-secreting β-cells are destroyed by immune cells that infiltrate the pancreatic islets (i.e., insulitis). Instead of preventing or reversing T1D, most treatments focus on alleviating symptoms with insulin-replacement therapy. Meanwhile, T1D is a heterogeneous disease that poses significant challenges to define mechanisms of pathogenesis and ultimately develop effective therapeutics. Using our chromatin-activity-based chemoproteomics (ChaC) to dissect T1D heterogeneity we have discovered a novel translational regulatory mechanism of T1D immunopathogenesis wherein G9a, a histone methyltransferase, noncanonically activates the translation of a battery of T1D-driving proteins. Further, we have deduced a mechanism of drug action wherein G9a inhibition, in nonobese diabetic (NOD) mice, a T1D model, mitigated β cell autoimmunity by specifcially suppressing the translation of T1D-related proteins in pathogenic effector T cells (Teff). Thus, we (TransChromix and UNC) will develop a new generation of mechanism-based, Teff-specific T1D therapeutics. Epidemiologic evidence showed that G9a is constitutively active in lymphocytes from T1D patients, implicating G9a-interacting pathways in T1D pathogenicity. Using ChaC with a biotinylated G9a inhibitor as a probe we captured and identified the same translation regulators that interact with G9a in both the NOD mice with highly infiltrated islets and in peripheral blood mononuclear cells (PBMCs) of T1D patients. Accordingly, we found that G9a inhibition or inhibition of Ezh2, a ChaC-identified G9a- interactor, in NOD mice, specifically reduced pancreas-infiltrating Teff that drive β cell autoimmunity. Further, quantitative proteomic analysis of inhibitor treated T1D mice revealed that G9a or Ezh2 inhibition downregulates in vivo expression of proteins regulating Teff pathogenicity, particularly those proteins related to glucose metabolism in diabetes, pancreatic disease pathway, and T cell proliferation. Importantly, the inhibitor-affected T1D proteome that represents clinical T1D pathology is mouse-to-human conserved, indicating that suppressing G9a-mediated, gene-specific translation in Teff cells is clinically practical for effective therapeutics of T1D. Because proteins directly mediate events promoting pathogenicity, we will test the hypothesis that targeting G9a- mediated translational mechanisms in autoreactive Teff provides an effective strategy to prevent and/or reverse T1D progression. In Phase I, NOD mice representing varying stages of disease progression will be treated with inhibitors, and we will (1) conduct studies to validate the in vivo Teff specificity of G9a or Ezh2 inhibitors, and to measure inhibitor toxicity, (2) determine the specificity and long-term efficacy of G9a or/and Ezh2 inhibition on NOD mice at early stages of T1D, and (3) for the human clinical validation we will determine by proteomic approaches the inhibitor effects on ex vivo cultures of the PBMCs from T1D patients. Our mechanistic discovery that G9a and Ezh2 regulate translation of proteins driving Teff-mediated b cell autoimmunity provides a new approach to treat T1D.

抽象的 1型糖尿病（T1D）是一种慢性自身免疫性疾病，其中分泌胰岛素的β细胞被破坏浸润胰岛的免疫细胞（即胰岛炎）。大多数疗法专注于减轻胰岛素替代疗法的症状。同时，T1D是异质的在定义发病机理机制并最终发展有效的疾病使用基于染色质活性的化学蛋白质组学（CHAC）剖析T1D异质性我们已经发现了一种新颖的T1D免疫病变的调节机制，其中G9a，A Hisstone甲基转移酶，非规范地激活了一组T1D驱动蛋白的翻译。更远，我们推导了一种药物作用的机制，其中g9a抑制在非肥胖糖尿病（NOD）小鼠中，T1D 模型，通过专门抑制T1D相关蛋白的翻译来缓解β细胞自身免疫性致病效应T细胞（TEFF）。那就是我们（Transchromix和UNC）将发展新一代基于机制的TEFF特异性T1D疗法。流行病学证据表明G9A一直是活跃于T1D患者的淋巴细胞中，暗示了T1D致病性中的G9A相互作用途径。使用CHAC 用生物素化的G9a抑制剂作为探针，我们捕获并确定了相同的翻译调节剂在两只小鼠中与高度浸润的胰岛和外周血单核细胞中的G9A相互作用 T1D患者的（PBMC）。根据以下内容，我们发现G9A抑制或抑制EZH2是CHAC识别的G9A- 相互作用的小鼠相互作用特异性降低了胰腺浸润的TEFF，驱动β细胞自身免疫性。更远，抑制剂处理的T1D小鼠的定量蛋白质组学分析表明，G9A或EZH2抑制作用下调调节TEFF致病性的蛋白质的体内表达，尤其是与葡萄糖有关的蛋白质糖尿病，胰腺疾病途径和T细胞增殖的代谢。重要的是，受抑制剂影响代表临床T1D病理学的T1D蛋白质组是鼠标到人类的保守，表明抑制 TEFF细胞中G9A介导的基因特异性翻译在临床上对于T1D的有效治疗是实用的。由于蛋白质直接介导促进致病性的事件，因此我们将检验以下假设：靶向G9A- 自动反应性TEFF中的介导的翻译机制提供了预防和/或反向的有效策略 T1D进展。在第一阶段，将用代表疾病进展的不同阶段的小鼠接受抑制剂，我们将（1）进行研究以验证G9A或EZH2抑制剂的体内TEFF特异性，以及测量抑制剂毒性，（2）确定G9A或/和EZH2抑制的特异性和长期有效性在T1D的早期阶段的NOD小鼠，（3）对于人类临床验证，我们将通过蛋白质组学确定接近抑制剂对T1D患者PBMC的离体培养物的影响。我们的机械发现 G9A和EZH2调节驱动TEFF介导的B细胞自动免疫的蛋白质的翻译可提供新的治疗T1D的方法。