地中海贫血症红细胞谱系发育分化基因筛查及机制研究

SCREENING AND MECHANISM OF ERYTHROID LINEAGE DIFFERENTIATION GENE IN THALASSEMIA

杨最

11849365

硕士

生物学

张健

2020-06-08

2020-06-08

哈尔滨工业大学

深圳

地中海贫血症是世界上最常见和发生率最高的一种单基因遗传病。目前临床上地中海贫血症没有可以普遍推广的根治方法。产前诊断在地中海贫血症的预防上有重要贡献。由于产前诊断技术的局限，导致地中海贫血症稀有突变类型不能被检测，不能完全避免地中海贫血症患儿出生。而外显子测序技术有可能弥补这一缺点，因此我们运用外显子测序技术对地中海贫血症基因携带者羊水样本进行检测，既能验证临床产前诊断结果，同时也是为了发现新的变异位点。我们通过外显子测序筛选出1344个共有变异位点。通过对这些共有变异位点进行GO富集分析筛选了26个term，涵盖膜的整体成分、跨膜信号受体活性、细胞外基质结构成分、钙离子结合等；通过KEGG富集分析筛选了13条信号通路，包括糖酵解/糖异生、ECM受体相互作用、补体与凝血级联反应等。这些基因和信号通路可能与地中海贫血症的发病机制密切相关，其结果需要进一步验证。另一方面，珠蛋白基因缺陷影响红细胞发育过程。红细胞的发育起源于造血干细胞，对造血干细胞的机理研究是根治地中海贫血症等一系列血液疾病的潜在突破口。我们利用磁珠分选和流式分选的方法从脐带血样本分离纯化CD34阳性细胞，在体外运用多种细胞因子成功对造血干细胞进行诱导分化，为体外培养造血干细胞模拟体内的造血过程奠定基础。

Thalassemia is one of the most common monogenic genetic diseases in the world. At present, there is no universal method to cure thalassemia. Prenatal diagnosis plays an important role in the prevention of thalassemia. Due to the limitation of prenatal diagnosis technology, the rare mutation type of thalassemia can not be detected and the birth of thalassemia children can not be completely avoided. Exon sequencing technology can make up for this shortcoming, so we use exon sequencing technology to detect amniotic fluid samples of Mediterranean anemia gene carriers, which can not only verify the clinical prenatal diagnosis results, but also to find new mutation sites. We screened 1344 common mutation sites by exon sequencing. Through GO enrichment analysis, 26 term were screened, including the whole membrane components, transmembrane signal receptor activity, extracellular matrix structural components, calcium binding, etc.; through KEGG enrichment analysis, 13 signal pathways were screened, including glycolysis / gluconeogenesis, ECM receptor interaction, complement and coagulation cascade, etc. These genes and signaling pathways may be closely related to the pathogenesis of thalassemia, and the results need to be further verified. On the other hand, hemoglobin gene defects affect the development of erythrocytes. The development of red blood cells originated from hematopoietic stem cells. The study of the mechanism of hematopoietic stem cells is a potential breakthrough to cure a series of blood diseases such as thalassemia. CD34 positive cells were isolated and purified from cord blood samples by magnetic bead sorting and flow sorting. In vitro, hematopoietic stem cells were successfully induced and differentiated by a variety of cytokines, which laid a foundation for in vitro culture of hematopoietic stem cells to simulate the hematopoietic process in vivo.

地中海贫血症 产前诊断 外显子测序 红细胞发育 造血调控 造血干细胞

Thalassemia prenatal diagnosis exon sequencing erythropoiesis hematopoietic regulation hematopoietic stem cells

中文

联合培养

南方科技大学

杨最. 地中海贫血症红细胞谱系发育分化基因筛查及机制研究[D]. 深圳. 哈尔滨工业大学,2020.