Dissecting the genetics of the Metabolic Syndrome on Chromosome 17 of the LH rat

描述（由申请人提供）：肥胖症、血脂异常、胰岛素抵抗、高血压等疾病统称为代谢综合征，是心血管疾病的主要危险因素。其发病率继续上升，部分原因是肥胖的流行增加。里昂高血压（LH）大鼠是一种强大的近交模型，用于解剖人类代谢综合征的遗传贡献，具有高体重，胆固醇，甘油三酯，胰岛素/葡萄糖比和盐敏感性高血压。有趣的是，同时从同一SD菌落中选择的里昂稳压（LN）对照菌株在遗传上与LH非常相似（85%），但在表型上却截然不同。在LH与LN杂交的遗传连锁定位中，鉴定出大鼠染色体（RNO） 17上的3个数量性状位点（QTL）簇与LH大鼠代谢综合征的多个特征有关，以及RNO17、2和13上的3个与血压有关的QTL。我们假设，LH染色体17具有与代谢综合征所有主要性状相关的基因，并且RNO 17上的QTL与RNO 17、2和13之间存在QTL相互作用。为了确定这些基因中的至少一个，我们提出了传统的定位克隆、计算机定位和QTL（表型和基因表达）定位相结合的方法来确定导致代谢综合征的致病基因和途径。我们将：1：验证LH和LN菌株RNO17的QTL区域。LH和LN菌株的密切遗传特性允许快速生成和表征经济LH- 17ln菌株和同源亚菌株。这些菌株将用于缩小QTL区域，并确定哪些表型是由多个QTL、相互作用QTL或QTL内单个基因的多效性效应引起的。2：研究反式和顺式单倍型在代谢综合征基础性状上的相互作用。我们将在LH和LN之间进行F2交叉，以确定RNO17、2和13上的QTL与导致我们在LH中观察到的表型的调控网络之间的相互作用。后代将通过表型分型、SNP基因分型和基因表达谱进行综合研究，以进行表型(p) QTL和表达(e) QTL定位。3：通过SNP精细定位和基因大鼠实验验证，完善RNO17的QTL。对于每个候选区域，我们将通过额外的SNP分型来精确定位单倍型，以缩小和定义最小的QTL间隔。所有的遗传变异将在这些最小区域确定候选基因和序列变异。这些区域将在同系学中进行实验验证。4：通过大鼠转基因救援验证功能变异。对于具有上述目标中鉴定的功能序列变异证据的最强候选，我们将开发转基因大鼠，将LN等位基因引入LH大鼠或将LH等位基因引入LH- 17ln。

DESCRIPTION (provided by applicant): The collection of maladies including obesity, dyslipidemia, insulin resistance and hypertension is referred to as the metabolic syndrome and is a major risk factor for cardiovascular disease. Its incidence continues to rise, in part as a result of the epidemic increase in obesity. The Lyon Hypertensive (LH) rat is a powerful inbred model for dissecting the genetic contributions of the human metabolic syndrome, with high body weight, cholesterol, triglycerides, and insulin/glucose ratios, and salt-sensitive hypertension. Interestingly, the Lyon normotensive (LN) control strain, concurrently selected from the same SD colony, is genetically quite similar (85%) to the LH, but phenotypically very distinct. Genetic linkage mapping in an LH x LN intercross identified 3 clusters of Quantitative Traits Loci (QTL) on rat chromosome (RNO) 17 for multiple features of the metabolic syndrome in the LH rat and 3 QTL for blood pressure on RNO17, 2, and 13. We hypothesize that the LH chromosome 17 has genes contributing to all major traits of the metabolic syndrome, and that there are QTL interactions between QTL on RNO 17 and between RNO 17, 2, and 13. To identify at least one of these genes, we propose a combined traditional positional cloning, in silico mapping, and QTL (phenotype and gene expression) mapping to identify causal genes and pathways leading to the metabolic syndrome. We will: 1: Validate the QTL regions for RNO17 in the LH and LN strains. The close genetic identity of the LH and LN strains allows for rapid generation and characterization of a consomic LH-17LN strain and congenic substrains. These strains will be used to narrow the QTL regions and to determine which phenotypes are due to multiple QTL, interacting QTL, or pleiotropic effects of a single gene within a QTL. 2: Investigate interactions between trans-acting and cis-acting haplotypes on traits underlying the metabolic syndrome. We will perform an F2 intercross between LH and LN to determine the interactions between QTL on RNO17, 2, and 13, and regulatory networks causing the phenotypes we observe in the LH. The offspring will be comprehensively studied by phenotyping, SNP genotyping, and gene expression profiling for combined phenotype (p) QTL and expression (e) QTL mapping. 3: Refine the QTL on RNO17 through SNP fine-mapping and experimental validation in congenic rats. For each candidate region, we will fine-map the haplotypes by additional SNP typing to narrow and define minimal QTL intervals. All genetic variation will be determined in these minimal regions to identify candidate genes and sequence variants. These regions will be experimentally validated in the congenics. 4: Validate functional variants through rat transgenic rescue. For the strongest candidate with evidence of functional sequence variants identified in the above aims, we will develop a transgenic rat, either introducing the LN allele into the LH rat or the LH allele into the LH-17LN.