利用分子表型遗传分析策略解析猪Th1/Th2内稳态平衡的遗传调控机制

Previous studies reported that the regulatory mechanism of Th1/Th2 homeostasis balance had an intense relationship with anti-disease ability through the active protective role when infection was occurring. We consider that the genetic regulatory mechanism of Th1/Th2 homeostasis can provide one of the important windows for revealing genetic basis of anti-disease capability of pigs. This project will use the experimental material and data from our previous studies to establish the molecular indicator traits of Th1/Th2 homeostasis through molecular phenotype strategy, with the purpose for translating Th1/Th2 homeostasis into measurable molecular phenotypes. We will develop a new method that can construct single-trait genetic network only using genotypic and phenotypic data. Using our new method, we will construct the genetic network of each indicator trait. Furthermore, we will analyze the corresponding relationship between network parameters and GWAS results after GWAS analysis, from which mine the important node genes and edges (i.e., gene pairs). And then, we will validate gene functions and regulatory relationships between gene pairs using the single cell-type transcriptomic analysis and in vitro cell immune stimulation experiments. Based on these results, we will clarify or partly clarify the genetic regulatory mechanism of Th1/Th2 homeostasis in pigs. This project will be helpful for understanding the genetic regulatory mechanism of anti-disease ability of pigs, promote the researches of pig anti-disease breeding, and also provide reference information for human clinical medicine.

Th1/Th2内稳态平衡在病原感染时发挥了积极的保护作用，与机体抗病力有着密切的关系。据此我们认为，猪Th1/Th2内稳态平衡遗传调控机制是揭示猪抗病力遗传基础的重要窗口。本项目拟在我室已积累的研究材料和数据的基础上，采用分子表型遗传分析策略，在系统构建Th1/Th2内稳态指示性状的基础上，应用新发展的基于基因型和表型信息的遗传网络构建法，构建各指示性状的遗传调控网络，系统评估遗传网络特性，联合GWAS结果与遗传网络参数，挖掘重要节点基因与边（即基因对），并通过单类细胞转录组、体外细胞免疫刺激实验等手段，验证基因功能与基因对的调控关系，藉此阐明猪Th1/Th2内稳态平衡的遗传调控机制。本项目实施后对于准确理解猪抗病力的遗传调控机制有重要帮助，将推动猪抗病育种研究的进一步发展，预期研究成果对人类临床医学研究也有一定的借鉴意义。

Th1/Th2是T淋巴细胞的重要亚群，为解析猪抗病力遗传基础提供了重要窗口。本项目从Th1/Th2细胞因子及其比率（内稳态）、T淋巴细胞单类（single-class）细胞转录组、单细胞（single-cell）转录组、重要免疫基因功能、免疫基因分析方法与工具开发等多角度开展了较为系统的研究，取得了较为系统的研究结果。主要研究结果包括：1）系统揭示了IL-10、IFN-γ、IL-4等Th1/Th2细胞因子及其比率的数量特征；2）利用单类细胞和单细胞转录组分析技术，揭示了T淋巴细胞分化的分子事件，构建了免疫基因互作网络，3）提出了偶联转录组新分析方法，建立了猪全基因组CRISPR敲除文库工具；4）从免疫多效性、功能突变等角度解析了ESR1、CD4、BIC/miR-155等重要免疫基因的功能；5）进一步对Th1/Th2细胞产物诱导的猪肺泡巨噬细胞极化机制、以及猪免疫性状的基因组选择问题开展了探索，提出sssGBLUP等方法，提高了基因组预测的准确性。本研究发表标注期刊论文5篇（其中SCI标注论文4篇），申报免疫基因分子标记国家发明专利5项（其中获授权发明专利2项），培养博士生、硕士生5名。本项目超额完成各项量化指标，总体上达成预期研究目标，研究获得的方法、工具和分子标记具有重要的应用前景。