Clinicopathological characteristics and genomic profile of primary sinonasal tract diffuse large B cell lymphoma (DLBCL) reveals gain at 1q31 and RGS1 encoding protein; high RGS1 immunohistochemical expression associates with poor overall survival in DLBCL not otherwise specified (NOS)

Raw annonymized data, OncoScan CEL files of the manuscript. PMID: 27775850 DOI: 10.1111/his.13106 ABSTRACT: <strong>Aims: </strong> We aimed to define the clinicopathological characteristics of 29 primary sinonasal diffuse large B cell lymphoma (DLBCL^sn ) in a series of 240 cases of DLBCL not otherwise specified [DLBCL^{all (}^NOS⁾ ], including DLBCL^sn training set (n = 11) and validation set (n = 18), and DLBCL^non-sn (n = 211). <strong>Methods and results: </strong> In the training set, 82% had a non-germinal center B-cell-like (Hans' Classifier) (non-GCB) phenotype and 18% were Epstein-Barr virus-encoded small RNAs (EBER)⁺ . The genomic profile showed gains⁽⁺⁾ of 1q21.3q31.2 (55%), 10q24.1 (46%), 11q14.1 (46%) and 18q12.1q23 (46%); losses^(-) of 6q26q27 (55%) and 9p21.3 (64%); and copy number neutral loss of heterozygosity (LOH) (acquired uniparental disomy, UPD) at 6p25.3p21.31 (36%). This profile is comparable to DLBCL^NOS (GSE11318, n = 203.) and closer to non-GCB/activated B-cell-like subtype (ABC). Nevertheless, +1q31, -9p21.3 and -10q11.1q26.2 were more characteristic of DLBCL^sn (P &lt; 0.001). Array results were verified successfully by fluorescence in situ hybridization (FISH) on +1q21.3 (CKS1B), -6q26 (PARK2), +8q24.21 (MYC), -9p21.3 (MTAP, CDKN2A/B), -17p13.1 (TP53) and +18q21.33 (BCL2) with 82-91% agreement. Minimal common regions included biologically relevant genes of MNDA (+1q23.1), RGS1 and RGS13 (+1q31.2), FOXP1 (+3p13), PRDM1 (BLIMP1) and PARK2 (-6q21q26), MYC (+8q24.21), CDKN2A (-9p21.3), PTEN (-10q23.31), MDM2 (+12q15), TP53 (-17p13.1) and BCL2 (+18q21.33). Correlation between DNA copy number and protein immunohistochemistry was confirmed for RGS1, RGS13, FOXP1, PARK2 and BCL2. The microenvironment had high infiltration of M2-like tumour associated macrophages (TAMs) and CD8⁺ T lymphocytes that associated with higher genomic instability. The DLBCL^sn validation set confirmed the clinicopathological characteristics, all FISH loci and immunohistochemistry (IHC) for RGS1. RGS1, one of the most frequently altered genes, was analysed by IHC in DLBCL^all and high RGS1 expression associated with non-GCB, EBER⁺ and unfavourable overall survival (hazard ratio = 1.794; P = 0.016). <strong>Conclusions: </strong> DLBCL^sn has a characteristic genomic profile. High RGS1 IHC expression associates with poor overall survival in DLBCL^{all (}^NOS⁾ .本研究手稿配套的经匿名化处理的原始数据，以及OncoScan CEL文件。PMID：27775850，DOI：10.1111/his.13106 摘要： <strong>研究目的：</strong> 本研究旨在明确240例非特指型弥漫大B细胞淋巴瘤（DLBCL not otherwise specified, DLBCL^{all (NOS)}）中29例原发鼻窦弥漫大B细胞淋巴瘤（sinonasal Diffuse Large B Cell Lymphoma, DLBCL^sn）的临床病理特征，研究队列包含DLBCL^sn训练集（n=11）、验证集（n=18）以及非鼻窦原发弥漫大B细胞淋巴瘤队列（DLBCL^non-sn，n=211）。 <strong>方法与结果：</strong> 训练队列中，82%的病例表现为非生发中心B细胞样（Hans分类法，non-germinal center B-cell-like, non-GCB）表型，18%的病例EB病毒编码小RNA（Epstein-Barr virus-encoded small RNAs, EBER）呈阳性。基因组特征显示存在1q21.3q31.2（55%）、10q24.1（46%）、11q14.1（46%）及18q12.1q23（46%）区域的拷贝数获得（+）；6q26q27（55%）与9p21.3（64%）区域的拷贝数缺失（-）；以及6p25.3p21.31区域的拷贝数中性杂合性缺失（copy number neutral loss of heterozygosity, LOH，即获得性单亲二体acquired uniparental disomy, UPD），发生率为36%。该基因组特征与非特指型弥漫大B细胞淋巴瘤（GSE11318，n=203）的特征相近，且更偏向非-GCB/活化B细胞样（activated B-cell-like, ABC）亚型。值得注意的是，+1q31、-9p21.3及-10q11.1q26.2是DLBCL^sn更为特征性的基因组改变（P < 0.001）。 本研究通过荧光原位杂交（fluorescence in situ hybridization, FISH）对以下位点进行了验证：+1q21.3（CKS1B）、-6q26（PARK2）、+8q24.21（MYC）、-9p21.3（MTAP、CDKN2A/B）、-17p13.1（TP53）以及+18q21.33（BCL2），验证一致性达82%-91%，结果均验证成功。最小共同受累区域包含多个具有生物学意义的基因：MNDA（+1q23.1）、RGS1与RGS13（+1q31.2）、FOXP1（+3p13）、PRDM1（BLIMP1）与PARK2（-6q21q26）、MYC（+8q24.21）、CDKN2A（-9p21.3）、PTEN（-10q23.31）、MDM2（+12q15）、TP53（-17p13.1）以及BCL2（+18q21.33）。本研究证实了DNA拷贝数与蛋白免疫组织化学（immunohistochemistry, IHC）表达水平的相关性，涉及RGS1、RGS13、FOXP1、PARK2及BCL2。肿瘤微环境中可见大量M2样肿瘤相关巨噬细胞（tumour associated macrophages, TAMs）与CD8⁺T淋巴细胞浸润，且该浸润与更高水平的基因组不稳定性相关。 DLBCL^sn验证队列进一步证实了其临床病理特征，所有FISH位点及RGS1的免疫组化结果均与前述发现一致。作为最常发生改变的基因之一，RGS1通过免疫组化在全部DLBCL^{all (NOS)}队列中进行了分析，结果显示高RGS1表达与non-GCB表型、EBER阳性及不良总生存期相关（风险比=1.794；P=0.016）。 <strong>结论：</strong> 原发鼻窦弥漫大B细胞淋巴瘤（DLBCL^sn）具有特征性的基因组谱。在全部非特指型弥漫大B细胞淋巴瘤（DLBCL^{all (NOS)}）中，RGS1的高免疫组化表达与不良总生存期显著相关。