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

基本信息

批准号：
10180992
负责人：
Chad Harmon Stahl
金额：
$ 31.16万
依托单位：
UNIV OF MARYLAND, COLLEGE PARK
依托单位国家：
美国
项目类别：
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/10180992
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 Industry Insulin Antagonists Insulin Resistance 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 fetal fetal programming genome wide association study high risk improved in utero insulin sensitivity insulin signaling metabolic profile muscle form non-invasive monitor obese mothers obesity development obesogenic offspring overexpression porcine model 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

项目摘要

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.

抽象的在人类和家畜中，成长中的胎儿经历代谢器官和细胞器的编程适应子宫内营养供应不足以及预期出生后也会出现类似的营养短缺。被称为“健康与疾病的发育起源（DOHaD）”或“节俭表型”的现象在人类和家畜中观察到，表现为“小于胎龄”(SGA) 或“低出生体重” (LBW) 后代在出生时在人类中，SGA 后代患肥胖症、糖尿病的风险很高。另一方面，低体重仔猪的饲料效率降低，生长性能和胴体特性发生改变，造成巨大的经济损失。 DOHaD 在猪模型中的作用，特别是具有天然节俭表型的 Ossabaw 猪，改善动物健康、产品质量和畜牧业盈利能力的潜在优势，同时同时作为人类的翻译模型，使其非常适合“双重目的” 使用这个模型，我们的主要目标是研究 GRB10（生长激素）的作用。受体结合蛋白 10)，一种调节人类 DOHaD 的衔接蛋白和胰岛素信号抑制剂。 GRB10 与 10% 患有严重 SGA 的 Silver–Russell 综合征 (SRS) 病例有关，并且根据 GWAS 研究在小鼠中，消融和过度表达实验表明 Grb10 是 II 型糖尿病的拮抗剂。胰岛素信号、生长抑制剂和代谢综合征的介质。我们的中心假设是胰岛素信号传导是介导节俭表型的关键，并且调节因此，GRB10 的表达将是克服 DOHaD 的关键，在本研究中，使用 CRISPR/Cas 系统，GRB10。 KO 和转基因敲入 (KI) Ossabaw 胎儿成纤维细胞与前体野生型 (WT) 一起产生。细胞、KO、KI 和 WT 仔猪的克隆系将通过体细胞核移植产生。实验管道中，Aim-1 将研究 GRB10 的消融和过度表达对产前和产后的影响。产后生长将在足月或产后生长一段时间后处死仔猪以评估其效果。 GRB10 缺失或过度表达对生长速度的影响 Aim-2 将研究胰岛素信号传导改变的影响。 GRB10 介导的代谢健康和胎儿编程的影响我们将研究 GRB10 的缺失或缺失。当喂养表现出肥胖饮食的动物时，过度表达对肥胖和糖尿病的发展有影响。将培育代谢综合征患者并研究其后代的跨代遗传我们预计该项目的结果将验证 GRB10 作为生长调节剂。农业上重要的性状，并作为糖尿病和代谢综合征治疗干预的候选者。