A porcine model for investigating the role of an insulin signaling regulator in development and disease

用于研究胰岛素信号调节剂在发育和疾病中的作用的猪模型

• 批准号: 9764398
• 负责人: Chad Harmon Stahl
• 金额: $ 31.63万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9764398
• 关键词: Ablation; Adaptor Signaling Protein; Adult; Age-Months; Agriculture; Animals; Binding Proteins; Birth; Birth Weight; Body Composition; Cardiovascular Diseases; Cells; Characteristics; Clustered Regularly Interspaced Short Palindromic Repeats; Coin; Development; Diabetes Mellitus; Diabetic mother; Diet; Disease; Domestic Animals; Dual-Energy X-Ray Absorptiometry; Economics; Epidemiology; Exhibits; Family Study; Family suidae; Fatty acid glycerol esters; Fetal Weight; Fetus; Fibroblasts; GRB10 gene; Generations; Glucose; Goals; Grant; Growth; Growth Hormone Receptor; Growth Inhibitors; Health; Human; Impairment; Insulin; Insulin Antagonists; Intramuscular; Investigation; Knock-in; Knock-out; Knockout Mice; Life; Link; Liver; Low Birth Weight Infant; Malnutrition; Meat; Mediating; Mediator of activation protein; Medicine; Metabolic; Metabolic Diseases; Metabolic syndrome; Mitochondria; Modeling; Mus; Muscle; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Nutritional; Nuts; Obesity; Organ; Organ Weight; Organelles; Pancreas; Performance; Phenotype; Placenta; Placental Insufficiency; Predisposition; Pregnancy; Production; Research; Risk Factors; Role; Silver-Russell syndrome; Small for Gestational Age Infant; Study models; Therapeutic Intervention; Time; Tissues; Transgenes; Transgenic Organisms; Visceral; Weaning; cohort; experimental study; feeding; fetal; fetal programming; genome wide association study; high risk; human model; improved; in utero; insulin sensitivity; insulin signaling; metabolic profile; muscle form; non-invasive monitor; obese mothers; obesity development; obesogenic; offspring; overexpression; postnatal; postnatal period; prenatal; prevent; programs; reduced muscle mass; response; somatic cell nuclear transfer; therapeutic candidate; trait; transgenerational epigenetic inheritance; translational model; translational study; western diet

ABSTRACT In humans and domestic animals, the growing fetus undergoes programming of metabolic organs and organelles to adapt for poor nutrient supply in utero and in anticipation of similar shortage of nutrients postnatally. This phenomenon known as “Developmental Origins of Health and Disease (DOHaD)” or “thrifty phenotype” is observed in humans and domestic animals, and manifests as “small for gestation age” (SGA) or “low birth weight” (LBW) offspring at the time of birth. In humans, the SGA offspring are at high risk for developing obesity, diabetes and cardiovascular diseases in postnatal life. The LBW piglets on the other hand exhibit reduced feed efficiency, growth performance and altered carcass characteristics, resulting in great economic losses. Therefore, the study of DOHaD in a pig model, specifically the Ossabaw pigs that have a naturally occurring thrifty phenotype has the potential advantage of improving animal health, product quality and profitability in animal agriculture, while simultaneously serving as a translational model for humans, making it an ideal fit for “Dual-purpose with dual- benefit” grant program. Using this model, our main goal is to investigate the role of GRB10 (growth hormone receptor binding protein 10), an adaptor protein and inhibitor of insulin signaling in mediating DOHaD. In humans, GRB10 is linked to 10% of Silver–Russell syndrome (SRS) cases with severe SGA, and from GWAS studies to Type II diabetes. In mice, ablation and overexpression experiments have highlighted Grb10 as an antagonist of insulin signaling, a growth inhibitor, and mediator of metabolic syndrome. Our Central hypothesis is that altered insulin signaling is key to mediating thrifty phenotype, and modulation of GRB10 expression will therefore be key to overcoming DOHaD. In this study, using CRISPR/Cas system, GRB10 KO and transgene knockin (KI) Ossabaw fetal fibroblasts were generated. Along with precursor wildtype (WT) cells, clonal lines of KO, KI, and WT piglets will be generated by somatic cell nuclear transfer. Using this experimental pipeline, Aim-1 will investigate the effect of ablation and overexpression of GRB10 in prenatal and postnatal growth. Piglets will be sacrificed at term or after a period of postnatal growth to evaluate the effect of loss or overexpression of GRB10 on growth rate. Aim-2 will investigate the effect of altered insulin signaling mediated by GRB10 on metabolic health and fetal programming. We will investigate the loss of GRB10 or overexpression on the development of obesity and diabetes when fed obesogenic diet. Animals that exhibit metabolic syndrome will be bred and the offspring investigated for transgenerational inheritance of the phenotype. We anticipate that the results from the project will validate GRB10 as a regulator of growth-an agriculturally important trait, and as a candidate for therapeutic intervention of diabetes and metabolic syndrome.

抽象的 在人类和家养动物中，日益增长的胎儿经历了代谢器官和细胞器的编程 适应子宫内营养不良的供应，并预期出生后的营养短缺。这 被称为“健康与疾病的发育起源（DOHAD）”或“节俭表型”的现象是 在人类和家畜中观察到，表现为“妊娠年龄较小”（SGA）或“低出生体重” （LBW）出生时后代。在人类中，SGA后代有肥胖症，糖尿病的高风险 以及产后生活中的心血管疾病。另一方面，LBW仔猪暴露了降低的进料效率， 增长性能和尸体特征改变，导致巨大的经济损失。因此，研究 猪模型中的多哈德（Dohad 改善动物健康，产品质量和在动物农业中的利润的潜在优势，而 同样，作为人类翻译模型，使其非常适合“双重用途，dual-dual- 赠款计划。使用此模型，我们的主要目标是调查GRB10的作用（增长本人） 受体结合蛋白10），一种介导DOHAD中胰岛素信号传导的衔接蛋白和抑制剂。在人类中 GRB10与严重SGA的10％的银 - 俄罗斯综合征（SRS）相关，从GWAS研究到 II型糖尿病。在小鼠中，消融和过表达实验突出了GRB10作为拮抗剂 胰岛素信号传导，一种生长抑制剂和代谢综合征的介体。 我们的中心假设是，改变的胰岛素信号传导是介导节俭表型的关键，并调节 因此，GRB10表达将是克服DOHAD的关键。在这项研究中，使用CRISPR/CAS系统，GRB10 KO和转化敲击蛋白（Ki）Ossabaw胎儿成纤维细胞。与前体野生型（WT）一起 细胞，KI，Ki和WT小猪的克隆线将通过体细胞核转移产生。使用此 实验管道，AIM-1将研究GRB10在产前和 产后生长。小猪将在期限或一段时间后生长后处死，以评估 GRB10对生长速率的损失或过表达。 AIM-2将研究胰岛素信号改变的效果 由GRB10介导有关代谢健康和胎儿编程的介导。我们将调查GRB10或 饲喂肥胖饮食时肥胖和糖尿病发育的过表达。展示的动物 代谢综合征将被育种，并研究了后代的转化遗传 表型。我们预计该项目的结果将验证GRB10作为增长的调节者 - 农业重要的特征，并作为糖尿病和代谢综合征治疗干预的候选人。