Viral Insulin-like Peptides (VILPs) and Their Activities on Mammalian Cells

病毒胰岛素样肽（VILP）及其对哺乳动物细胞的活性

• 批准号: 9810855
• 负责人: Emrah Altindis
• 金额: $ 11.15万
• 依托单位: BOSTON COLLEGE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9810855
• 关键词: 3T3-L1 Cells; Acute; Adipocytes; Affect; Affinity; Apoptosis; Binding; Bioinformatics; Blood; Blood Glucose; Bombyx; Bombyx mori; Caenorhabditis elegans; Cell Line; Cell Proliferation; Cells; Cellular Metabolic Process; Chronic; Collaborations; Complementary DNA; Country; Cysteine; DNA; DNA biosynthesis; Data; Dependovirus; Diabetes Mellitus; Disease; Distant; Double Stranded DNA Virus; Drosophila genus; Drosophila melanogaster; Family; Fibroblasts; Fishes; Functional disorder; Gene Expression; Genes; Genome; Gluconeogenesis; Goals; Growth; Hepatocyte; High Fat Diet; Histology; Hormones; Human; IGF1 gene; Injections; Insulin; Insulin Receptor; Insulin-Dependent Diabetes Mellitus; Insulin-Like Growth Factor I; Insulin-Like Growth Factor II; Insulin-Like-Growth Factor I Receptor; Invertebrates; Iridovirus; Knowledge; Laboratories; Ligands; Liver; Longevity; Lower Organism; Mammalian Cell; Metabolism; Mission; Molecular; Mus; Names; Nematoda; Non-Insulin-Dependent Diabetes Mellitus; Organism; Pathogenesis; Peptide Signal Sequences; Peptides; Phylogenetic Analysis; Physiological; Physiology; Prevention therapy; Process; Protein Isoforms; Relaxin; Research; Resistance; Rodent; Role; Serum; Signal Transduction; Singapore; Skin; Snail Venoms; Somatomedins; Specificity; Streptozocin Diabetes; Stress; Structure; System; Testing; Time; Tissues; Translating; United States National Institutes of Health; Vertebrates; Viral; Viral Physiology; Virus; Virus Diseases; Work; adeno-associated viral vector; adipocyte differentiation; analog; animal tissue; autocrine; blood glucose regulation; cell growth; design; disulfide bond; experimental study; glucose uptake; growth promoting activity; human disease; in vivo; innovation; insulin signaling; lipid biosynthesis; member; novel; overexpression; paracrine; peptidomimetics; receptor; transcriptome sequencing; tumor; tumor growth

PROJECT SUMMARY/ABSTRACT Using a bioinformatics analysis, we have identified viral insulin/IGF1-like peptides (VILPs) in the genomes of four different, but related, viruses. Although these viruses were isolated from fish, assessment of the VILPs in a phylogenetic context showed that VILPs are equally well-related to humans and other species as to fish insulins/IGFs. Nothing is known about how these VILPs might interact with mammalian insulin or IGF-1 receptors, and about the potential impact of these viruses and their insulin-like peptides in terms of diabetes pathogenesis or organismal/tumor growth, all processes highly regulated by mammalian insulins and insulin- like growth factors. The overarching goal of this proposal is to functionally characterize the VILPs, define their mechanism of action, understand their potency and determine if they affect mammalian pathophysiology. The central hypothesis is that VILPs are new members of the insulin super-family that can interact with mammalian insulin and IGF-1 receptors and activate insulin/IGF-1 signaling thereby altering cellular metabolism, gene expression and cell proliferation. This hypothesis has been formulated on the basis of exciting preliminary data produced in the applicant's laboratory. The rationale for the proposed research is that understanding the functions of VILPs on mammalian cells has the potential to translate into better understanding of fundamental mechanisms and early origins of insulin/IGF-1 signaling, and how these peptides may be involved in not only viral diseases, but also type 2 diabetes (T2D), type 1 diabetes (T1D) and conditions of tumor growth. Together these disorders affect millions of people in this country and worldwide. Guided by our preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Determine the molecular mechanisms of VILP action as novel insulin and IGF-1 receptor ligands and 2) explore the pathophysiologic effects of VILPs on glucose homeostasis and cell growth in mice. Under Aim 1, functional characterization of VILPs will be performed using human, rodent and fish cells. In addition to exogenous effects, VILPs will be cloned into mammalian cells and their effects as endogenous ligands will be assessed. With collaborators, the NMR structures of VILPs will be determined and compared to their mammalian counterparts. Under Aim 2, we will characterize the acute and chronic effects of VILPs in vivo. To this end, VILP genes will be transferred to the liver of mice using an adeno-associated viral vector system. The local effects of VILP overexpression on liver and their systemic effects on the body will be determined This approach is innovative since it will be the first exploration of VILP action and their impact on insulin signaling, metabolism, gene expression and growth. The proposed research is significant, because it will be the first study that advances our understanding of VILP function and their potential impact on human disease. Ultimately, such knowledge has the potential to help us not only understand the role of these molecules, but to better understand insulin/IGF-1 action overall, which may be useful in designing new, unique insulin analogs.

项目摘要/摘要 使用生物信息学分析，我们已经确定了在基因组中的病毒胰岛素/IGF1样肽（VILP） 四种不同但相关的病毒。尽管这些病毒是从鱼类中分离出来的 系统发育环境表明，维尔普斯与人类和其他物种相关，而不是 胰岛素/IGFS。对于这些Vilps如何与哺乳动物胰岛素或IGF-1相互作用，尚不了解 受体，以及这些病毒及其胰岛素样肽在糖尿病方面的潜在影响 发病机理或生物/肿瘤生长，所有过程都受哺乳动物胰岛素和胰岛素 - 喜欢增长因素。该提案的总体目标是在功能上表征vilps，定义其 作用机理，了解其效力并确定它们是否影响哺乳动物的病理生理学。这 中心假设是VILP是可以与哺乳动物相互作用的胰岛素超级家族的新成员 胰岛素和IGF-1受体并激活胰岛素/IGF-1信号传导，从而改变细胞代谢，基因 表达和细胞增殖。该假设是根据令人兴奋的初步数据提出的 在申请人的实验室中生产。拟议研究的理由是了解 VILP在哺乳动物细胞上的功能有可能转化为更好地理解基本的 胰岛素/IGF-1信号的机制和早期起源，这些肽不仅可能参与 病毒疾病，但还包括2型糖尿病（T2D），1型糖尿病（T1D）和肿瘤生长状况。一起 这些疾病会影响这个国家和世界范围内数以百万计的人。在我们的初步数据的指导下， 假设将通过追求两个具体目的来检验：1）确定VILP的分子机制 作为新型胰岛素和IGF-1受体配体的作用，以及2）探索VILPS的病理生理影响 小鼠的葡萄糖稳态和细胞生长。在AIM 1下，Vilps的功能表征将是 使用人，啮齿动物和鱼细胞进行。除了外源作用外，VILP还将克隆到 将评估哺乳动物细胞及其作为内源配体的作用。与合作者，NMR Vilps的结构将被确定并与其哺乳动物对应物进行比较。在AIM 2下，我们将 表征体内维尔普斯的急性和慢性作用。为此，VILP基因将被转移到 使用腺相关病毒载体系统的小鼠肝脏。 VILP过表达对肝脏的局部影响 他们对身体的系统影响将确定这种方法是创新的，因为它将是第一个 VILP作用及其对胰岛素信号传导，代谢，基因表达和生长的影响。这 拟议的研究很重要，因为这将是我们对VILP理解的第一个研究 功能及其对人类疾病的潜在影响。最终，这种知识有可能帮助我们 不仅了解这些分子的作用，而且可以更好地了解胰岛素/IGF-1总体上的作用，这是 设计新的独特的胰岛素类似物可能很有用。