β cell miRNAs Function as Molecular Hubs of Type 1 Diabetes Pathogenesis

β 细胞 miRNA 作为 1 型糖尿病发病机制的分子中心

• 批准号: 10561855
• 负责人: Carmella Evans-Molina
• 金额: $ 4.09万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10561855
• 关键词: Affect; Beta Cell; Biological Markers; Biology; Biometry; Body mass index; Categories; Cell Proliferation; Cell Survival; Cell physiology; Cells; Clinical; Clinical Research; Collaborations; Data; Data Science; Disease; Enrollment; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Gestational Diabetes; Glucose Intolerance; Health; Human; Hyperglycemia; Insulin; Insulin Resistance; Insulin-Dependent Diabetes Mellitus; Metabolic; Methods; MicroRNAs; Modeling; Molecular; Monitor; Mothers; Mus; Non-Insulin-Dependent Diabetes Mellitus; Nulliparity; Obesity; Oral; Outcome Study; Overweight; Pathway interactions; Physicians; Plasma; Play; Pregnancy; Pregnancy Outcome; Pregnant Women; Process; Research; Role; Sampling; Scientist; Second Pregnancy Trimester; Structure of beta Cell of islet; Testing; Time; Training; Untranslated RNA; Up-Regulation; Writing; career; circulating microRNA; diabetes pathogenesis; diabetes risk; experimental study; health data; improved; insulin secretion; islet; miRNA expression profiling; offspring; potential biomarker; skills; transcriptome sequencing; translational study

ABSTRACT: Gestational diabetes mellitus (GDM) is a heterogenous disease that is defined by the occurrence of glucose intolerance or hyperglycemia during the late second trimester and occurs when pancreatic β cells cannot secrete sufficient insulin when there is increasing insulin resistance. GDM affects approximately 10% of all pregnancies and negatively impacts the short-and long-term health of both pregnant mothers and their offspring. microRNAs (miRNAs, 18-25 nt) are a class of small non-coding RNAs that post transcriptionally modulate gene expression and have been shown to regulate several key processes within the β cell. miRNAs have also been implicated as potential biomarkers for type 1 diabetes, and type 2 diabetes. Past studies have also demonstrated a potential for miRNAs to serve as biomarkers for GDM as well, but most only employed targeted approaches and did not consider the contribution of maternal overweight/obesity. Against this background, we obtained plasma samples collected from pregnant mothers enrolled in the Nulliparous Pregnancy Outcomes Study-Monitoring Mothers-to-be (nuMoM2b; NCT01322529) study to perform unbiased miRNA sequencing. Our preliminary data showed that maternal overweight/obesity status (defined as BMI≥25) influenced plasma miRNA signatures. Compared to controls in the same BMI category, we identified a set of miRNAs that were different in pregnant mothers who subsequently developed GDM. This result indicates the potential of miRNA signatures to predict GDM onset prior to onset of hyperglycemia. Interestingly, we also found that miR-517a-3p, a placental-enriched miRNA that is known to increase cellular proliferation, was downregulated in mothers who subsequently developed GDM. In Aim 1, we hypothesize that the upregulation of miR-517a-3p positively regulates β cell function and survival. To test this hypothesis, we will define the unreported role of miR-517a-3p in regulating β cell function using INS-1 cells, murine islets, and human islet models, as well as using state-of-the-art RNA-seq approach to uncover the modulated genes and molecular pathways. In Aim 2, we hypothesize that a combination of miRNA signatures and clinical variables will improve the prediction of GDM. This hypothesis will be tested by quantifying the expression levels of these miRNA signatures using expanded sample sets from the multi-center nuMoM2b study and analyzed using appropriate model-selection methods in collaboration with Dr. Joanne Daggy (IU Dept of Biostatistics and Health Data Science). This R01 supplement will also support time for the candidate (Dr. Kua) to perform experiments and attend training courses to acquire new research skills to pursue a career as an independent physician scientist. The candidate training will include three main objectives: (1) Obtain state-of-the-art training in miRNA biology and their role in modulating β cell function and gestational diabetes risk, (2) develop experimental toolkits to support translational and clinical studies in gestational diabetes and offspring metabolic programming, and (3) refine scientific dissemination skills, including oral presentation and scientific writing skills.

摘要：妊娠糖尿病（GDM）是一种由此定义的异源性疾病 第二个妊娠期葡萄糖肠道或高血糖症，发生胰腺β细胞时发生 当胰岛素耐药性增加时，不能秘密促进足够的胰岛素。 GDM影响约10％ 所有怀孕和负面影响怀孕的母亲及其长期健康 后代。 microRNA（mirnas，18-25 nt）是一类小型非编码RNA，在转录后 调节基因表达，并已证明可以调节β细胞内的几个关键过程。 mirnas 也被认为是1型糖尿病和2型糖尿病的潜在生物标志物。过去的研究有 还证明了miRNA也有可能作为GDM的生物标志物，但大多数员工也有可能 有针对性的方法，没有考虑孕产妇超重/肥胖的贡献。反对这一点 背景，我们获得了从nulliparous入学的怀孕母亲那里收集的血浆样本 妊娠结局研究监测到准妈妈（NUMOM2B； NCT01322529）进行无偏见的研究 miRNA测序。我们的初步数据表明，母体超重/肥胖状态（定义为BMI≥25） 影响血浆miRNA签名。与同一BMI类别中的对照相比，我们确定了一组 随后发展GDM的怀孕母亲不同的miRNA。该结果表明 miRNA特征在高血糖发作之前预测GDM发作的潜力。有趣的是，我们也是如此 发现miR-517a-3p是一种富含胎盘的miRNA，已知会增加细胞增殖，是 后来发展GDM的母亲的母亲下调。在AIM 1中，我们假设更新 miR-517A-3P的阳性调节β细胞功能和存活。为了检验这一假设，我们将定义 miR-517a-3p在调节β细胞功能中使用INS-1细胞，鼠胰岛和人类胰岛的未报告作用 模型，以及使用最先进的RNA-Seq方法来发现调制基因和分子 途径。在AIM 2中，我们假设miRNA特征和临床变量的组合将改善 GDM的预测。该假设将通过量化这些miRNA的表达水平进行检验 使用来自多中心NUMOM2B研究的扩展样品集的签名，并使用适当的 模型选择方法与Joanne Daggy博士合作（生物统计学和健康数据IU部 科学）。此R01补充剂还将支持候选人（KUA博士）执行实验的时间 参加培训课程，以获得新的研究技能，从而从事独立的物理科学家职业。 候选人培训将包括三个主要目标：（1）获得miRNA生物学的最先进培训 以及它们在调节β细胞功能和妊娠糖尿病风险中的作用，（2）开发实验工具包 支持妊娠糖尿病和后代代谢编程中的翻译和临床研究，（3） 完善科学传播技巧，包括口头表现和科学写作技巧。