Division of Molecular Life Sciences, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
Immunogenetics. 2013 Jun;65(6):417-27. doi: 10.1007/s00251-013-0691-z. Epub 2013 Mar 9.
Graft-versus-host disease (GVHD) is the main complication after hematopoietic stem cell transplantation (HSCT). Evidence for non-HLA gene polymorphisms as a cause of GVHD lacks consistency, which is, in part, due to methodological issues of previous candidate gene association studies and small effect size of their results, demanding for larger scale and more robust approaches. Here, non-HLA gene polymorphisms were studied on a large population (922 HSCT pairs) from a homogeneous ethnic background with selection/correction for important clinical confounders. A methodology was applied exploiting the strength of confirmatory typing in an independent study cohort. Targeting an immunogenome of 2,909 genes, an approach of pooled DNA typing of 4,321 microsatellite (MS) markers in two independent screening steps and confirmation of associated markers by further individual genotyping on combined screening cohorts was used to identify genetic susceptibility loci for moderate to severe GVHD (grades 2-4). Ten MS loci (D5S424, D6S0035i, D1S0818i, DXS0151i, D17S0219i, DXS0629i, DXS0324i, D17S0271i, D6S0330i, and D1S1335i) passed the two pooled DNA typing steps and confirmation by individual sample genotyping; two of these (D1S0818i-ELTD1 and D6S0035i-MAPK14) remain associated following application of Bonferroni's correction and multivariate analysis. The MAPK14 locus was exemplarily explored by typing of haplotype single nucleotide polymorphisms (SNP) confirming this association. This study identified several new MS susceptibility loci for GVHD that warrant further investigation. Immunogenome scanning using MS markers is a useful method for the identification of non-HLA gene loci associating with HSCT outcomes.
移植物抗宿主病（GVHD）是造血干细胞移植（HSCT）后的主要并发症。非 HLA 基因多态性作为 GVHD 病因的证据缺乏一致性，这在一定程度上是由于先前候选基因关联研究的方法学问题和结果的小效应量，需要更大规模和更稳健的方法。在这里，在具有重要临床混杂因素选择/校正的同质人群中（922 对 HSCT），对非 HLA 基因多态性进行了大规模研究。应用了一种方法，该方法利用了在独立研究队列中确证性分型的优势。针对免疫基因组 2909 个基因，通过两个独立的筛选步骤对 4321 个微卫星（MS）标记进行 pooled DNA 分型，并在联合筛选队列中对相关标记进行进一步的个体基因分型以确认，用于确定中度至重度 GVHD（2-4 级）的遗传易感性位点。通过 pooled DNA 分型的两个步骤和个体样本基因分型的确认，有 10 个 MS 位点（D5S424、D6S0035i、D1S0818i、DXS0151i、D17S0219i、DXS0629i、DXS0324i、D17S0271i、D6S0330i 和 D1S1335i）通过了检测；其中两个（D1S0818i-ELTD1 和 D6S0035i-MAPK14）在应用 Bonferroni 校正和多变量分析后仍然存在相关性。通过对单核苷酸多态性（SNP）的单体型分型，对 MAPK14 基因座进行了探索，证实了这种关联。本研究鉴定了几个新的与 GVHD 相关的 MS 易感基因座，值得进一步研究。使用 MS 标记进行免疫基因组扫描是一种鉴定与 HSCT 结果相关的非 HLA 基因座的有用方法。
