Long non-coding RNAs in Islet Cell Biology

胰岛细胞生物学中的长非编码 RNA

• 批准号: 9212938
• 负责人: LORI SUSSEL
• 金额: $ 38.88万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9212938
• 关键词: Adult; Affect; Alleles; Alpha Cell; Antisense RNA; B-Cell Development; Beta Cell; Biological; Biological Preservation; Biology; Cell Line; Cell physiology; Cells; Cellular biology; Code; Collection; Complex; Computational algorithm; Data; Development; Diabetes Mellitus; Disease; Endocrine; Future; Gene Expression Profiling; Gene Expression Regulation; Generations; Genes; Genetic Transcription; Genome; Glucagon; Goals; Human; In Vitro; Insulin; Insulin-Dependent Diabetes Mellitus; Islet Cell; Islets of Langerhans; Knockout Mice; Knowledge; Lead; Link; Maintenance; Messenger RNA; Molecular; Mouse Strains; Mus; Non-Insulin-Dependent Diabetes Mellitus; Nucleotides; Onset of illness; Outcome Study; Pancreas; Pathogenesis; Pathway interactions; Phenotype; Play; Population; Proteins; Regulatory Pathway; Research; Role; Signal Pathway; Single Nucleotide Polymorphism Map; Somatic Cell; Source; Specificity; Stem cells; Synteny; Techniques; Testing; Tissues; Transcript; Treatment Efficacy; Untranslated RNA; Work; base; blood glucose regulation; comparative; endocrine pancreas development; experimental study; genome wide association study; genome-wide analysis; human disease; islet; knock-down; mammalian genome; neuroblastoma cell; novel; null mutation; pancreas development; prevent; therapeutic target

A large number of research efforts are currently underway to understand and prevent diabetes. Towards this goal, studies in mice have significantly advanced our understanding of the conserved signaling pathways and regulatory factors required for the development and maintenance of functional β cells. However, it is evident from the current challenges associated with predicting and treating diabetes, and generating alternative sources of endocrine cells, that we are still missing key molecular components that are required to generate, mature and preserve fully functional β cells. Recently, advances in genome biology have revealed that a large part of the mammalian genome is transcribed and includes a large number of long non-coding RNAs (lncRNAs), many which are conserved between mouse and human. LncRNAs are a diverse group of transcripts longer than 200 nucleotides that resemble mRNAs, but do not encode proteins. There is emerging evidence to suggest lncRNAs may be involved in β cell function and diabetes. In support of this data, it has been determined that the majority of diabetes-associated SNPs map to non-coding regions of the genome, many that are specifically located in lncRNAs. Therefore, the identification and characterization of conserved islet-specific lncRNAs will significantly further our understanding of the regulatory networks that control β cell development and function, and how disruption of these mechanisms may lead to diabetes. For this study, we have used comparative gene expression analyses and novel computational algorithms to identify and rank potentially relevant lncRNAs expressed in the human and mouse pancreas. We have selected two of these lncRNAs for further functional analysis based on a defined set of criteria, including their stage and level of expression, islet specificity, synteny and conservation between mouse and human, proximity to pancreas-related coding genes, and proximity to SNPs associated with diabetes. The overall goal of this proposal is to reveal the functional roles of selected lncRNAs in islet and β cell biology. We hypothesize that long non-coding RNAs (lncRNAs) represent a missing component of the islet transcriptional regulatory pathways and play essential roles in β cell development and function. To test this hypothesis, we will characterize the function and molecular activity of βlinc1, a novel lncRNA expressed in mouse and human islets. We have generated mice that are deleted for βlinc1 and will characterize the βlinc1 KO phenotype and determine the molecular activity of βlinc1. We have also identified Paupar as a conserved lncRNA that is highly expressed in adult glucagon-expressing α cells. In addition, Paupar regulates Pax6 expression and a number of Pax6-independent and dependent transcriptional activities. We have generated a null allele of Paupar by inserting an H2B-GFP cassette into the Paupar locus. We propose to characterize the expression, functional role and molecular mechanism of Paupar in the pancreatic islet.

目前正在进行大量的研究工作，以了解和预防糖尿病。为了实现这一目标，对小鼠的研究显着提高了我们对功能性β细胞开发和维持所需的配置信号通路和调节因素的理解。然而，这是从与预测和治疗糖尿病以及生成内分泌细胞替代来源相关的当前挑战中的证据，我们仍然缺少生成，成熟和保留功能齐全的β细胞所需的关键分子成分。最近，基因组生物学的进步表明，哺乳动物基因组的很大一部分被转录，其中包括大量长的非编码RNA（LNCRNA），许多RNA（LNCRNA）在小鼠和人之间保守。 lncRNA是一组较长的转录本的潜水员群，比200个类似于mRNA但不编码蛋白质的核动体。有新兴的证据表明LNCRNA可能参与β细胞功能和糖尿病。为了支持该数据，已经确定大多数与糖尿病相关的SNP映射到基因组的非编码区域，许多图像专门位于LNCRNA中。因此，构成的特异性LNCRNA的鉴定和表征将大大进一步了解控制β细胞发育和功能的调节网络，以及这些机制的破坏如何导致糖尿病。在这项研究中，我们使用了比较基因表达分析和新颖的计算算法来识别和排除在人和小鼠胰腺中表达的潜在相关的LNCRNA。我们已经选择了这些LNCRNA中的两个进行进一步的功能分析，以定义的一组标准，包括它们的表达阶段和水平，小鼠与人之间的表达，胰岛特异性，同步和保护，与胰腺相关的编码基因的接近，以及与糖尿病相关的SNP的接近。该提案的总体目标是揭示选定的LNCRNA在胰岛和β细胞生物学中的功能作用。我们假设长的非编码RNA（LNCRNA）代表了胰岛转录调节途径的缺失成分，并且在β细胞发育和功能中起着重要作用。为了检验这一假设，我们将表征我们已经产生的小鼠的功能和分子活性，这些小鼠被删除了βlinc1，并将表征βlinc1KO表型并确定βlinc1的分子活性。我们还确定了Paupar为构成的lncRNA，在表达成人胰高血糖素的α细胞中。此外，Paupar调节PAX6表达和许多与PAX6无关的和依赖的转录活性。我们通过将H2B-GFP盒式盒子插入Paupar基因座来产生了Paupar的无效等位基因。我们建议表征Paupar在胰岛中的表达，功能和分子机制。