Forget B G, Degan B A, Arcasoy M O
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
Trans Am Clin Climatol Assoc. 2000;111:38-44; discussion 44-5.
We studied a kindred with dominantly inherited familial erythrocytosis associated with heterozygosity for a deletion of seven nucleotides in exon 8 of the EpoR gene resulting in an EpoR peptide that is truncated by 59 amino acids in its C-terminal intracytoplasmic signal transduction domain. A seven basepair direct repeat sequence is present in the normal EpoR gene at the site of this mutation, consistent with the slipped mispairing model for the generation of short deletions during DNA replication. Hypersensitivity to erythropoietin of primary erythroid progenitors from an affected individual was observed in in vitro cultures of peripheral blood mononuclear cells, as indicated by the growth, at suboptimal concentrations of added Epo, of more numerous and larger BFU-E-derived erythroid cell colonies compared to those obtained from a normal control subject. To study mutant EpoR function, the cDNA encoding the mutant EpoR was stably transfected into murine growth factor (IL-3)-dependent 32D tissue culture cells. In proliferation assays, cells expressing the mutant EpoR displayed 5 to 10-fold increased sensitivity to Epo compared to cells expressing similar numbers of the wild type EpoR. In addition, the cells transfected with the mutant truncated receptor demonstrated prolonged activity of Jak2 kinase and Stat5 activity, molecules that mediate signal transduction by the EpoR.
我们研究了一个具有显性遗传的家族性红细胞增多症家系,该家系与促红细胞生成素受体(EpoR)基因第8外显子中7个核苷酸缺失的杂合性相关,导致EpoR肽在其C端胞质内信号转导结构域中被截短59个氨基酸。在正常EpoR基因的该突变位点存在一个7个碱基对的直接重复序列,这与DNA复制过程中短缺失产生的滑动错配模型一致。在受影响个体的外周血单个核细胞体外培养中,观察到原代红系祖细胞对促红细胞生成素的超敏反应,表现为在添加的促红细胞生成素浓度低于最佳浓度时,与正常对照个体相比,来自更多且更大的爆式红系集落形成单位(BFU-E)的红系细胞集落生长。为了研究突变型EpoR的功能,将编码突变型EpoR的cDNA稳定转染到依赖鼠生长因子(IL-3)的32D组织培养细胞中。在增殖试验中,与表达相似数量野生型EpoR的细胞相比,表达突变型EpoR的细胞对促红细胞生成素的敏感性提高了5至10倍。此外,用突变型截短受体转染的细胞显示Jak2激酶和Stat5活性延长,这些分子介导EpoR的信号转导。