Voon Hsiao Phin Joanna, Wardan Hady, Vadolas Jim
Cell and Gene Therapy Research Group, The Murdoch Childrens Research Institute, The University of Melbourne, Royal Children's Hospital, Flemington Road, Parkville 3052, Melbourne, Australia.
Blood Cells Mol Dis. 2007 Sep-Oct;39(2):184-8. doi: 10.1016/j.bcmd.2007.01.006. Epub 2007 May 9.
Beta-thalassaemia is an inherited disease caused by defective synthesis of the beta-globin chain of haemoglobin, leading to an imbalance in globin chains. Excess alpha-globin chains precipitate in erythroid progenitor cells resulting in cell death, ineffective erythropoiesis and severe anaemia. Decreased alpha-globin synthesis leads to milder symptoms, exemplified by individuals who co-inherit alpha-thalassaemia and beta-thalassaemia. In this study, we set out to investigate whether co-inheritance of alpha- and beta-thalassaemia in mice leads to reduced anaemia. Heterozygous murine beta-globin knockout (KO) mice (beta+/-) which display severe anaemia were mated with heterozygous alpha-globin KO mice (alpha++/--). The resulting progeny were genotyped and classed as wild-type WT (alpha++/++;beta+/+), heterozygous alpha-KO (alpha++/--;beta+/+), heterozygous beta-KO (alpha++/++;beta+/-) or double heterozygous (DH) alpha-KO/beta-KO (alpha++/--;beta+/-). Mice were bled and full blood examinations (FBE) performed. FBE results for heterozygous beta-KO mice (beta+/-) showed marked reductions in haemoglobin and haematocrit levels and significant increases in red cell distribution widths and reticulocyte counts compared to WT mice. In contrast, FBE results for DH alpha-KO/beta-KO mice showed near normal red blood cell indices. These results indicate that reduction of alpha-globin expression leads to correction of the globin chain imbalance in beta-thalassaemic mice and therefore an improved phenotype. The analysis of DH alpha-KO/beta-KO mice leads to the following conclusions: (1) co-inheritance of alpha- and beta-thalassaemia in mice improves the thalassaemic phenotype, identical to the situation in humans; (2) the heterozygous murine beta-globin KO mouse model is a suitable in vivo model to test for therapeutic knockdown of alpha-globin.
β地中海贫血是一种遗传性疾病,由血红蛋白β珠蛋白链合成缺陷引起,导致珠蛋白链失衡。过量的α珠蛋白链在红系祖细胞中沉淀,导致细胞死亡、无效造血和严重贫血。α珠蛋白合成减少会导致症状较轻,例如同时遗传α地中海贫血和β地中海贫血的个体。在本研究中,我们着手调查小鼠中α和β地中海贫血的共同遗传是否会减轻贫血症状。将表现出严重贫血的杂合子小鼠β珠蛋白敲除(KO)小鼠(β+/-)与杂合子α珠蛋白KO小鼠(α++/--)交配。对产生的后代进行基因分型,并分类为野生型WT(α++/++;β+/+)、杂合子α-KO(α++/--;β+/+)、杂合子β-KO(α++/++;β+/-)或双杂合子(DH)α-KO/β-KO(α++/--;β+/-)。对小鼠进行采血并进行全血细胞检查(FBE)。与WT小鼠相比,杂合子β-KO小鼠(β+/-)的FBE结果显示血红蛋白和血细胞比容水平显著降低,红细胞分布宽度和网织红细胞计数显著增加。相比之下,DHα-KO/β-KO小鼠的FBE结果显示红细胞指数接近正常。这些结果表明,α珠蛋白表达的降低可纠正β地中海贫血小鼠的珠蛋白链失衡,从而改善表型。对DHα-KO/β-KO小鼠的分析得出以下结论:(1)小鼠中α和β地中海贫血的共同遗传可改善地中海贫血表型,与人类情况相同;(2)杂合子小鼠β珠蛋白KO小鼠模型是测试α珠蛋白治疗性敲低的合适体内模型。