Circulating miRNAs and prediction of beta-cell treatment response: The Restoring Insulin Secretion Study

摘要 尽管接受了治疗，但相当大比例的2型糖尿病(T2D)患者并未经历临床 改善(即缺乏治疗反应)。在分析恢复胰岛素分泌(RISE)的数据时 这项研究是一项随机对照试验，测试内科和外科治疗改善和 为了维持β细胞的功能，我们团队的成员在治疗中表现出了显著的异质性 在β细胞水平上使用高血糖钳夹金标准测量的反应。我们知道的很少， 然而，关于T2D治疗反应异质性的影响因素以及缺乏前瞻性的工具 确定治疗应答者和无应答者。造成这种情况的一个主要原因是缺乏可循环使用的生物标志物 在β细胞水平上预测或反映治疗反应。葡萄糖重要基因的表观遗传调控 而通过诸如microRNAs(MiRNAs)等短的非编码RNA(MiRNAs)进行的胰岛素代谢可能介导改善或 治疗过程中β细胞功能的保留和循环中的miRNA可能是治疗的预测生物标志物 回应。在β细胞中，miRNA对基因表达的调节对胰岛素的分泌和细胞都是至关重要的。 营业额。此外，分泌到循环中的miRNAs可能反映了组织水平的表达差异。 和/或积极参与器官间相声。而在各种组织和循环中的miRNAs 已被证明可通过暴露于人类或体外模型中的RISE干预而改变，其影响 这些在β细胞中的miRNA大部分还没有被描述。使用Rise儿科杂志的数据和样本 药物研究(n=49；R03DK122100；PI WANDER)，我们确定了9个具有前瞻性的血浆miRNAs 与10-19岁青年的治疗反应有关，包括两个新的miRNAs，miR-4468和miR-6727， 它们被预测通过Bcl2样蛋白1和胰岛淀粉样多肽来调节胰岛细胞的凋亡。我们的目标是 成年成人β细胞功能维持或改善相关miRNA的特征 受试者，通过钳夹衍生的β细胞反应与胰岛素敏感性的测量来定义，在治疗后 用药物或胃束带治疗。为了实现这一目标，我们提出了一项由两部分组成的研究。在目标1中，我们将分析 在嵌套病例对照中基线和治疗后使用下一代测序的循环miRNAs 治疗应答者与无应答者的样本上升。在目标2中，我们将检查miR-4468、miR- 6727，以及Aim 1中鉴定的miRNA对β-细胞功能、存活和功能增益中去分化的影响 在人类胰岛中使用锁定的基于核酸的miRNA模拟的体外实验。我们最重要的假设是 循环中的miRNA通过影响胰岛素分泌和β细胞存活来促进治疗的异质性。 据我们所知，这个项目将是第一个直接测试与治疗相关的miRNAs在调节或 反映治疗反应。更好地理解miRNAs作为治疗异质性的生物标志物将 通过前瞻性地确定可能从特定治疗中受益的亚组，促进T2D精准医学 并促进制定有针对性的干预措施，以防止或延缓T2D的进展。

Abstract Despite treatment, a significant proportion of patients with type 2 diabetes (T2D) do not experience clinical improvement (i.e., lack treatment response). In analyses of data from the Restoring Insulin Secretion (RISE) Study, a randomized controlled trial testing the potential of medical and surgical therapies to improve and maintain β-cell function, members of our team have demonstrated substantial heterogeneity in treatment response at the level of the β cell using gold standard measures from hyperglycemic clamps. We know little, however, about contributing factors to heterogeneity of T2D treatment response and lack tools to prospectively identify treatment responders and non-responders. A major reason for this is lack of circulating biomarkers that predict or reflect treatment response at the level of the β cell. Epigenetic regulation of genes important in glucose and insulin metabolism by short non-coding RNAs such as microRNAs (miRNAs) may mediate improvement or preservation of β-cell function with treatment, and circulating miRNAs may be predictive biomarkers of treatment response. In the β cell, regulation of gene expression by miRNAs is crucial to both insulin secretion and cell turnover. In addition, miRNAs secreted into the circulation may reflect tissue-level differences in expression and/or participate actively in inter-organ crosstalk. While miRNAs in a variety of tissues and in the circulation have been shown to be altered by exposure to the RISE interventions in humans or in vitro models, effects of most of these miRNAs in the β cell are not yet described. Using data and samples from the RISE Pediatric Medication Study (n=49; R03DK122100; PI Wander), we identified 9 plasma miRNAs that are prospectively related to treatment response in youth aged 10–19 years including two novel miRNAs, miR-4468 and miR-6727, which are predicted to regulate islet apoptosis via Bcl-2 like 1 and islet amyloid polypeptide. Our objective is to characterize miRNAs that are related to preservation or improvement in β-cell function among adult RISE participants, defined by clamp-derived measures of β-cell response paired with insulin sensitivity, after treatment with medications or gastric banding. To achieve this goal, we propose a two-part study. In Aim 1, we will assay circulating miRNAs using next-generation sequencing at baseline and after treatment in a nested case-control sample of treatment responders vs. non-responders in RISE. In Aim 2, we will examine effects of miR-4468, miR- 6727, and miRNAs identified in Aim 1 on β-cell function, survival, and de-differentiation in gain-of-function in vitro experiments using locked nucleic acid-based miRNA mimics in human islets. Our overarching hypothesis is that circulating miRNAs contribute to treatment heterogeneity via effects on insulin secretion and β-cell survival. To our knowledge, this project will be the first to directly test the role of treatment-related miRNAs in mediating or reflecting treatment response. Better understanding of miRNAs as biomarkers of treatment heterogeneity will facilitate T2D precision medicine by prospectively identifying subgroups likely to benefit from specific treatments and promote development of targeted interventions to prevent or delay the progression of T2D.