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）是一种异质性疾病， 在妊娠中期晚期发生葡萄糖耐受不良或高血糖，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%的 对孕妇及其子女的短期和长期健康产生负面影响。 后代microRNAs（miRNAs，18-25 nt）是一类小的非编码RNA，在转录后 调节基因表达，并已显示调节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 也证明了miRNAs作为GDM生物标志物的潜力，但大多数仅用于 没有考虑到产妇超重/肥胖的影响。反对这个 在背景下，我们获得了从参加Nulliparous研究的孕妇中收集的血浆样品。 妊娠结局研究-监测准妈妈（nuMoM 2b; NCT 01322529）研究，以进行无偏倚 miRNA测序。我们的初步数据显示，母亲超重/肥胖状态（定义为BMI≥25） 影响血浆miRNA签名。与相同BMI类别的对照组相比，我们发现了一组 这些miRNAs在随后发生GDM的孕妇中是不同的。这个结果表明 在高血糖症发作之前预测GDM发作的miRNA特征的潜力。有趣的是，我们也 发现miR-517 a-3 p，一种胎盘富集的miRNA，已知可以增加细胞增殖， 在随后发生GDM的母亲中下调。在目标1中，我们假设上调 miR-517 a-3 p的表达正调节β细胞功能和存活。为了验证这个假设，我们将定义 使用INS-1细胞、鼠胰岛和人胰岛研究miR-517 a-3 p在调节β细胞功能中的作用 模型，以及使用最先进的RNA-seq方法来揭示调节的基因和分子 途径。在目标2中，我们假设miRNA标签和临床变量的组合将改善 GDM的预测这一假设将通过定量这些miRNA的表达水平来检验。 使用来自多中心nuMoM 2b研究的扩展样本集的特征，并使用适当的 与Joanne Daggy博士（IU生物统计和健康数据系）合作的模型选择方法 科学）。该R 01补充材料还将支持候选人（Kua博士）进行实验的时间， 参加培训课程，以获得新的研究技能，追求作为一个独立的医生科学家的职业生涯。 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 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， 支持妊娠期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，包括口头陈述和科学写作技能。