Chou Stella T, Flanagan Jonathan M, Vege Sunitha, Luban Naomi L C, Brown R Clark, Ware Russell E, Westhoff Connie M
Division of Hematology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA.
Division of Hematology, Department of Pediatrics, Baylor College of Medicine, Houston, TX.
Blood Adv. 2017 Aug 3;1(18):1414-1422. doi: 10.1182/bloodadvances.2017007898. eCollection 2017 Aug 8.
genes are highly polymorphic and encode the most complex of the 35 human blood group systems. This genetic diversity contributes to Rh alloimmunization in patients with sickle cell anemia (SCA) and is not avoided by serologic Rh-matched red cell transfusions. Standard serologic testing does not distinguish variant Rh antigens. Single nucleotide polymorphism (SNP)-based DNA arrays detect many and variants, but the number of alleles tested is limited. We explored a next-generation sequencing (NGS) approach using whole-exome sequencing (WES) in 27 Rh alloimmunized and 27 matched non-alloimmunized patients with SCA who received chronic red cell transfusions and were enrolled in a multicenter study. We demonstrate that WES provides a comprehensive genotype, identifies SNPs not interrogated by DNA array, and accurately determines zygosity. Among this multicenter cohort, we demonstrate an association between an altered genotype and Rh alloimmunization: 52% of Rh immunized vs 19% of non-immunized patients expressed variant Rh without co-expression of the conventional protein. Our findings suggest that allele variation in patients with SCA is clinically relevant, and NGS technology can offer a comprehensive alternative to targeted SNP-based testing. This is particularly relevant as NGS data becomes more widely available and could provide the means for reducing Rh alloimmunization in children with SCA.
基因具有高度多态性，编码人类35种血型系统中最复杂的一种。这种遗传多样性导致镰状细胞贫血（SCA）患者发生Rh同种免疫，且血清学Rh匹配的红细胞输血无法避免。标准血清学检测无法区分变异的Rh抗原。基于单核苷酸多态性（SNP）的DNA阵列可检测到许多 和 变异，但所检测的等位基因数量有限。我们采用全外显子组测序（WES）的新一代测序（NGS）方法，对27例发生Rh同种免疫和27例匹配的未发生同种免疫的SCA患者进行了研究，这些患者均接受慢性红细胞输血且参与了一项多中心研究。我们证明WES可提供全面的 基因型，识别DNA阵列未检测的SNP，并准确确定 纯合性。在这个多中心队列中，我们证明 基因型改变与Rh同种免疫之间存在关联：52%的Rh免疫患者与19%的未免疫患者表达变异Rh，而无传统蛋白的共表达。我们的研究结果表明，SCA患者的 等位基因变异具有临床相关性，NGS技术可为基于靶向SNP的检测提供全面的替代方法。随着NGS数据越来越广泛可得，这一点尤为重要，并且可为减少SCA儿童的Rh同种免疫提供手段。
