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： 妊娠期糖尿病（GDM）是一种异质性疾病，其定义为： 妊娠中期后期葡萄糖不耐受或高血糖的发生，当胰腺 β 细胞 当胰岛素抵抗增加时，不能分泌足够的胰岛素。 GDM 影响大约 10% 所有怀孕都会对怀孕母亲及其子女的短期和长期健康产生负面影响 后代。 microRNA（miRNA，18-25 nt）是一类小非编码 RNA，在转录后 调节基因表达，并已被证明可以调节 β 细胞内的几个关键过程。 miRNA 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 也证明了 miRNA 作为 GDM 生物标志物的潜力，但大多数只使用 有针对性的方法，没有考虑母亲超重/肥胖的影响。反对这个 背景，我们获得了从参加未产儿的怀孕母亲那里收集的血浆样本 怀孕结果研究 - 监测准妈妈（nuMoM2b；NCT01322529）研究以执行公正 miRNA 测序。我们的初步数据显示，产妇超重/肥胖状态（定义为BMI≥25） 影响血浆 miRNA 特征。与相同 BMI 类别的对照相比，我们确定了一组 随后患上 GDM 的孕妇的 miRNA 有所不同。这一结果表明 miRNA 特征在高血糖发作之前预测 GDM 发作的潜力。有趣的是，我们还 发现 miR-517a-3p（一种富含胎盘的 miRNA，已知可增加细胞增殖） 在随后患有 GDM 的母亲中下调。在目标 1 中，我们假设上调 miR-517a-3p 正向调节 β 细胞功能和存活。为了检验这个假设，我们将定义 未报道的 miR-517a-3p 在使用 INS-1 细胞、鼠胰岛和人胰岛调节 β 细胞功能中的作用 模型，以及使用最先进的 RNA-seq 方法来揭示受调节的基因和分子 途径。在目标 2 中，我们假设 miRNA 特征和临床变量的结合将改善 GDM的预测。该假设将通过量化这些 miRNA 的表达水平来检验 使用来自多中心 nuMoM2b 研究的扩展样本集进行签名，并使用适当的方法进行分析 与 Joanne Daggy 博士（印第安纳大学生物统计和健康数据系）合作的模型选择方法 科学）。此 R01 补充品还将支持候选人（Kua 博士）进行实验和 参加培训课程以获得新的研究技能，以从事独立医师科学家的职业生涯。 候选人培训将包括三个主要目标：(1) 获得最先进的 miRNA 生物学培训 及其在调节 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规划的转化和临床研究，以及 (3) 提高科学传播技能，包括口头表达和科学写作技能。