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: 10441485
• 负责人: Mohamed Salem
• 金额: $ 31.17万
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10441485
• 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

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.

摘要 在人类和家畜中，生长中的胎儿经历代谢器官和细胞器的编程 以适应子宫内营养供应不足和预期出生后类似的营养缺乏。这 被称为“健康和疾病的发育起源”或“节俭表型”的现象是 在人类和家畜中观察到，并表现为“小于妊娠年龄”（SGA）或“低出生体重” (LBW)出生时的孩子。在人类中，SGA的后代患肥胖症、糖尿病的风险很高， 和心血管疾病。另一方面，LBW仔猪表现出降低的饲料效率， 生长性能和胴体特性的改变，造成巨大的经济损失。因此研究 在猪模型中，特别是具有天然存在的节俭表型的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， 同时作为人类的翻译模型，使其成为“两用”的理想选择。 福利”补助计划。利用这个模型，我们的主要目标是研究GRB 10（生长激素）的作用， 受体结合蛋白10），一种衔接蛋白和胰岛素信号传导的抑制剂，介导DOHaD。在人类中， GRB 10与10%的严重SGA的Silver-Russell综合征（SRS）病例相关，从GWAS研究中， II型糖尿病在小鼠中，消融和过表达实验突出了Grb 10作为一种拮抗剂， 胰岛素信号传导、生长抑制剂和代谢综合征的介质。 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， 因此，GRB 10表达将是克服DOHaD的关键。本研究利用CRISPR/Cas系统， 产生KO和转基因敲入（KI）Ossabaw胎儿成纤维细胞。沿着前体野生型（WT） 细胞，KO、KI和WT仔猪的克隆系将通过体细胞核转移产生。使用此 Aim-1将研究GRB 10的消融和过表达在产前和产后的影响。 出生后的成长将在足月时或出生后生长一段时间后处死仔猪，以评价 GRB 10的缺失或过表达对生长速率的影响。Aim-2将研究胰岛素信号传导改变的影响 通过GRB 10介导的代谢健康和胎儿编程。我们将调查GRB 10的损失， 过度表达对肥胖和糖尿病的发展的影响。动物展示 将繁殖代谢综合征，并调查后代的跨代遗传， 表型我们预计，该项目的结果将验证GRB 10作为生长调节剂的有效性， 农业上重要的性状，并作为糖尿病和代谢综合征的治疗干预的候选者。

项目成果

Extraembryonic Endoderm (XEN) Cells Capable of Contributing to Embryonic Chimeras Established from Pig Embryos.

• DOI: 10.1016/j.stemcr.2020.11.011
• 发表时间: 2021-01-12
• 期刊: Stem cell reports
• 影响因子: 5.9
• 作者: Park CH;Jeoung YH;Uh KJ;Park KE;Bridge J;Powell A;Li J;Pence L;Zhang L;Liu T;Sun HX;Gu Y;Shen Y;Wu J;Izpisua Belmonte JC;Telugu BP
• 通讯作者: Telugu BP