Amaro A, Parodi F, Diedrich K, et al.
单核苷酸 葡萄膜 乳脂
重要性：染色体6p扩增与葡萄膜黑色素瘤（UMs）的更多良性行为相关，而Ums具有3号染色体单倍体授予的转移高风险。染色体6P包含免疫调节基因B7家族的几个成员，包括嗜乳脂蛋白2（BTNL2；OMIM，606000），这是与前列腺癌的风险和自身免疫性疾病例。目的：调查UM患者6号染色体扩增候选基因BTNL2的表达和等位基因变异频率。设计，设置和参与者：在这个病例对照研究中，我们分析了UM细胞系和UM患者巨噬细胞内BTNL2的表达。BTNL2变异通过聚合酶链反应和高分辨率熔解使用探针分析。在209例UM患者和116相匹配的对照组患者分析以及12个UM和64其他肿瘤细胞系中分析错义突变rs28362679和rs41441651与肿瘤风险的相关性。通过芯片鉴定111例UM患者M1和M2型巨噬细胞中表达差异的基因，用Cox回归方程分析这些基因的表达与无病生存的相关性。资料收集于2013九月至2015年11月。.主要评价指标：嗜乳脂蛋白2单核苷酸变异与UM风险相关；M1和M2巨噬细胞特异性基因的表达与无病生存相关。结果：我们的基因型共325例。在209例UM患者中, 124（59.3%）为男性，114（54.5%）是意大利人，95（45.5%）是德国；平均年龄为65岁（范围）（27-94）年。意大利控制的116个患者中，67（57.8%）是女性，且平均年龄为39岁（范围）（21-88）年。嗜乳脂蛋白2是在UM和巨噬细胞中表达的。和欧洲变异存档和外显子聚生体数据2134/118564 [ 1.8% ]；95% CI，1.7-1.9）及外显子测序项目数据（100/4540 [ 2.2% ]；95% CI，1.8-2.7）相比，UM患者rs28362679变异频率更高（16 209 [ 7.7% ]；95% CI，4.7-12.2），而与意大利对照患者相比（10/116 [ 8.6% ]；95% CI，4.6-15 4）不高。目前rs41441651 变异出现在5例UM患者(2.4%; 95% CI, 0.9-5.7)，2例意大利对照患者(1.7%; 95% CI, 0.1-6.5), 2846例来自欧洲的变异与外显子聚集联盟数据档案患者（2.4%；95% CI，2.3-2.5），和23例外显子测序项目数据的患者（0.5%；95% CI 0.3-0.8）。人类UM细胞表达M1和M2巨噬细胞特异性基因，其表达与无病生存相关。结论与相关性：嗜乳脂蛋白2，在UM细胞和巨噬细胞表达水不同平，可能干扰肿瘤的免疫控制。嗜乳脂蛋白2变异在种族相关人群中表现出很高的可变频率。与对照者相比，UM患者没有表现出BTNL2变异富集。
Importance:Chromosome 6p amplification is associated with more benign behavior for uveal melanomas (UMs) with an otherwise high risk of metastasis conferred by chromosome 3 monosomy. Chromosome 6p contains several members of the B7 family of immune regulator genes, including butyrophilin-like 2 (BTNL2; OMIM, 606000), which is associated with prostate cancer risk and autoimmune diseases.Objective:To investigate the expression and variant allele frequencies of BTNL2, a candidate gene for chromosome 6 amplification, in patients with UM.Design, Setting, and Participants:In this case-control study, we analyzed the expression of BTNL2 in UM cell lines and human macrophages in patients with UM. Variants of BTNL2 were analyzed using probes for polymerase chain reaction and high-resolution melting. The association of missense variants rs28362679 and rs41441651 with tumor risk was analyzed in 209 patients with UM and 116 matched control patients as well as 12 UM and 64 other tumor cell lines. Genes that were differentially expressed in M1- and M2-polarized macrophages were identified by microarray analysis of 111 patients with UM, and the association of the expression of these genes with disease-free survival was analyzed by Cox regression analysis. Data were collected from September 2013 to November 2015.Main Outcomes and Measures:Butyrophilin-like 2 single-nucleotide variants were associated with UM risk; M1 and M2 macrophage-specific gene expression was associated with disease-free survival.Results:We genotyped a total of 325 patients. Of the 209 patients with UM, 124 (59.3%) were male, 114 (54.5%) were Italian, and 95 (45.5%) were German; the mean (range) age was 65 (27-94) years. Of the 116 Italian control patients, 67 (57.8%) were female, and the mean (range) age was 39 (21-88) years. Butyrophilin-like 2 is expressed in patients with UM and macrophages. The frequency of the rs28362679 variant was higher in patients with UM (16 of 209 [7.7%]; 95% CI, 4.7-12.2) than frequencies from European Variation Archive and Exome Aggregation Consortium data (2134 of 118564 [1.8%]; 95% CI, 1.7-1.9) and Exome Sequencing Project data (100 of 4540 [2.2%]; 95% CI, 1.8-2.7) but were not higher compared with Italian control patients (10 of 116 [8.6%]; 95% CI, 4.6-15.4). The rs41441651 variant was present in 5 patients with UM (2.4%; 95% CI, 0.9-5.7), 2 Italian control patients (1.7%; 95% CI, 0.1-6.5), 2846 patients from European Variation Archive and Exome Aggregation Consortium data (2.4%; 95% CI, 2.3-2.5), and 23 patients from Exome Sequencing Project data (0.5%; 95% CI, 0.3-0.8). Human UM cells express M1 and M2 macrophage-specific genes, whose expression is associated with disease-free survival.Conclusions and Relevance:Butyrophilin-like 2, expressed at various levels by UM cells and macrophages, might interfere with the immune control of the tumor. Butyrophilin-like 2 variants showed highly variable frequencies among ethnically related cohorts. There was no enrichment of BTNL2 variants in patients with UM compared with control patients.