Development of an early diagnostic biomarker and novel treatment strategy for T1D

开发 T1D 早期诊断生物标志物和新治疗策略

• 批准号: 9980379
• 负责人: Anath Shalev
• 金额: $ 53.76万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-15 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9980379
• 关键词: Ablation; Adult; Affect; Antihypertensive Agents; Apoptosis; Area Under Curve; Beta Cell; Biological Markers; Biology; C-Peptide; Calcium Channel; Calcium Channel Blockers; Cell Death; Cell physiology; Clinic; Clinical; Clinical Trials; Data; Deltastab; Detection; Development; Diabetes Mellitus; Diagnosis; Diphtheria Toxin; Disease; Disease Marker; Disease Progression; Dose; Down-Regulation; Drug Kinetics; Early Diagnosis; Exposure to; Gene Expression; Genes; Genetic; Glucose; Goals; Human; Individual; Inflammatory; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans; Measures; Mediating; MicroRNAs; Modeling; Molecular; Monitor; Mus; Oral; Oral Administration; Pathway interactions; Patients; Pharmaceutical Chemistry; Pharmacology; Placebos; Process; Production; Property; Randomized; Reagent; Regimen; Regulation; Resources; Risk; Sampling; Serum; Staging; Streptozocin; Structure; Structure of beta Cell of islet; TXNIP gene; Testing; Therapeutic; Time; Verapamil; Xenograft procedure; base; clinical diagnostics; cytokine; diagnostic biomarker; double-blind placebo controlled trial; high throughput screening; human RNA sequencing; humanized mouse; improved; in vivo; inhibitor/antagonist; insulin dependent diabetes mellitus onset; islet; mouse model; novel; potential biomarker; protective effect; response; small molecule; targeted treatment; tool; transcriptome sequencing; treatment strategy

Loss of insulin producing beta cells is a major problem of T1D and neither therapies that target this process nor simple measures/biomarkers that quantify it are currently available in humans. We recently discovered thioredoxin-interacting protein (TXNIP) as an attractive target in this regard, since genetic or pharmacological TXNIP downregulation with verapamil reduced beta cell apoptosis, increased pancreatic beta cell mass, and protected against and even reversed diabetes in mice. Now our, randomized, double-blind, placebo-controlled trial revealed that oral verapamil added for 12 months to a standard insulin regimen in adult subjects with recent onset T1D improved mixed meal-stimulated C-peptide area under the curve as a measure of endogenous beta cell function. While these findings have the advantage that they can be readily implemented into the clinic, they also underline the need for more specific tools to inhibit TXNIP. In fact, a novel, non-toxic, small molecule TXNIP inhibitor has now shown great promise in mouse models and human islets exposed to T1D-associated inflammatory cytokines. We further discovered that a specific TXNIP-induced microRNA is not only the most highly enriched microRNA in human beta cells, but is also released from dying beta cells, detectable in mouse and human serum, increased in T1D, and inversely correlated with remaining beta cell function. We therefore hypothesize that TXNIP inhibition provides a novel treatment strategy for T1D and that this serum microRNA represents an early diagnostic biomarker for T1D-associated beta cell loss in humans. To test this hypothesis we propose the following Specific Aims: #1: Study the effects of TXNIP inhibition as a novel treatment strategy for T1D including (i) test the effects of oral inhibitor administration on humanized mice using xenotransplantation of human islets into immunedeficient NRG-RIP-DTR mice to assess how TXNIP inhibition affects human islets in vivo in the context of diabetes and (ii) determine how the inhibitor functions and affects islet biology at a molecular level by elucidating the changes in gene expression and pathways involved using our reagents pertaining to TXNIP regulation and RNA sequencing of treated human islets. #2: Assess serum microRNA changes as an early diagnostic biomarker for T1D-associated beta cell loss. We will further explore our serum microRNA as a biomarker for T1D-associated beta cell loss during (i) disease progression as well as (ii) in response to verapamil-induced protection of beta cell function, taking advantage of our unique resource of pre-collected T1D serum samples from our clinical verapamil trial and (iii) will investigate the possibility that increased serum microRNA levels might predate the onset of T1D and could be used as an early marker for disease in at-risk subjects. Consistent with the goals of this HINR-CBDS RFA, the proposed studies will help develop a novel early T1D treatment strategy and a clinical diagnostic tool for the detection and staging of recently-diagnosed individuals with T1D.

产生β细胞的胰岛素丧失是T1D的主要问题，并且既不针对此过程的疗法，也不是对其量化其量化的疗法，目前在人类中可用。我们最近在这方面发现了硫氧还蛋白相互作用的蛋白（TXNIP）是一个有吸引力的靶标，因为遗传或药理学TXNIP下调，维拉帕米降低了β细胞细胞凋亡，胰腺β细胞量增加，并受到小鼠中甚至逆转糖尿病的保护。现在，我们的，随机的，双盲的，安慰剂对照试验表明，口服Verapamil在成年受试者的标准胰岛素方案中添加了12个月，最近发作T1D改善了策展下的混合餐食刺激的C肽面积，以作为内源性β细胞功能的量度。尽管这些发现的优势是可以轻松地将其实施到诊所中，但它们也强调了更具体的工具来抑制TXNIP。实际上，一种新型，无毒的小分子TXNIP抑制剂现已在暴露于T1D相关的炎性细胞因子的小鼠模型和人类胰岛中表现出巨大的希望。我们进一步发现，特定的TxNIP诱导的microRNA不仅是人β细胞中最富集的microRNA，而且还从垂死的β细胞中释放出来，在小鼠和人血清中可检测到T1D，在T1D中增加，并且与剩余β细胞功能成反比。因此，我们假设TXNIP抑制为T1D提供了一种新的治疗策略，并且该血清microRNA代表了人类与T1D相关β细胞损失的早期诊断生物标志物。为了检验这一假设，我们提出以下具体目的：＃1：研究TxNIP抑制作用作为T1D的新型治疗策略，包括（i）测试口服抑制剂给药对人类胰岛的人性化小鼠的影响，使用人类胰岛对人性化的植入进行人性化小鼠的影响，使用人类胰岛对免疫的NRG-RIP-DRIP-DTRES进行ii txnip的影响。通过阐明使用与TXNIP调控和RNA测序有关的人类胰岛的试剂的变化，确定抑制剂如何在分子水平上的功能并影响分子水平的胰岛生物学。 ＃2：评估血清microRNA的变化是T1D相关β细胞损失的早期诊断生物标志物。我们将进一步探索我们的血清microRNA作为（I）疾病进展过程中与T1D相关的β细胞损失的生物标志物，以及（ii）响应Verapamil诱导的β细胞功能的保护，利用我们在预测的T1D Serum样品中的独特资源，并将其persitation pertation pertate pertate pertate pertate pertation properation properatation可能增加，这可能会增加（iii）。作为高危受试者疾病的早期标记。与此Hinr-CBD RFA的目标一致，拟议的研究将有助于制定一种新型的早期T1D治疗策略和一种临床诊断工具，用于检测和分期最近被诊断为T1D的个体。