Purpose: Diffuse large B-cell lymphoma (DLBCL) is molecularly and clinically heterogeneous, and can be subtyped according to genetic alterations, cell-of-origin, or microenvironmental signatures using high-throughput genomic data at the DNA or RNA level. Although high-throughput proteomic profiling has not been available for DLBCL subtyping, MYC/BCL2 protein double expression (DE) is an established prognostic biomarker in DLBCL. The purpose of this study is to reveal the relative prognostic roles of DLBCL genetic, phenotypic, and microenvironmental biomarkers. Experimental Design: We performed targeted next-generation sequencing; IHC for MYC, BCL2, and FN1; and fluorescent multiplex IHC for microenvironmental markers in a large cohort of DLBCL. We performed correlative and prognostic analyses within and across DLBCL genetic subtypes and MYC/BCL2 double expressors. Results: We found that MYC/BCL2 double-high-expression (DhE) had significant adverse prognostic impact within the EZB genetic subtype and LymphGen-unclassified DLBCL cases but not within MCD and ST2 genetic subtypes. Conversely, KMT2D mutations significantly stratified DhE but not non-DhE DLBCL. T-cell infiltration showed favorable prognostic effects within BN2, MCD, and DhE but unfavorable effects within ST2 and LymphGen-unclassified cases. FN1 and PD-1–high expression had significant adverse prognostic effects within multiple DLBCL genetic/phenotypic subgroups. The prognostic effects of DhE and immune biomarkers within DLBCL genetic subtypes were independent although DhE and high Ki-67 were significantly associated with lower T-cell infiltration in LymphGen-unclassified cases. Conclusions: Together, these results demonstrated independent and additive prognostic effects of phenotypic MYC/BCL2 and microenvironment biomarkers and genetic subtyping in DLBCL prognostication, important for improving DLBCL classification and identifying prognostic determinants and therapeutic targets.

中文翻译：

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免疫微环境和 MYC/BCL2 双表达的遗传亚型和表型特征揭示了弥漫性大 B 细胞淋巴瘤的异质性

目的：弥漫性大 B 细胞淋巴瘤 (DLBCL) 在分子和临床上具有异质性，可以使用 DNA 或 RNA 水平的高通量基因组数据根据遗传改变、细胞起源或微环境特征进行亚型分类。尽管高通量蛋白质组学分析尚未可用于 DLBCL 亚型分析，但 MYC/BCL2 蛋白双表达 (DE) 是 DLBCL 中既定的预后生物标志物。本研究的目的是揭示 DLBCL 遗传、表型和微环境生物标志物的相对预后作用。实验设计：我们进行了靶向二代测序；MYC、BCL2 和 FN1 的 IHC；以及针对一大群 DLBCL 中微环境标志物的荧光多重 IHC。我们对 DLBCL 基因亚型和 MYC/BCL2 双表达基因内部和之间进行了相关性和预后分析。结果：我们发现 MYC/BCL2 双高表达 (DhE) 在 EZB 遗传亚型和 LymphGen 未分类的 DLBCL 病例中具有显着的不良预后影响，但在 MCD 和 ST2 遗传亚型中没有。相反，KMT2D 突变使 DhE 显着分层，但对非 DhE DLBCL 没有显着分层。T 细胞浸润在 BN2、MCD 和 DhE 中显示出有利的预后作用，但在 ST2 和 LymphGen 未分类病例中显示出不利的影响。FN1 和 PD-1 高表达在多个 DLBCL 遗传/表型亚组中具有显着的不良预后影响。尽管 DhE 和高 Ki-67 与 LymphGen 未分类病例中较低的 T 细胞浸润显着相关，但 DhE 和 DLBCL 遗传亚型中的免疫生物标志物的预后作用是独立的。结论：总的来说，这些结果证明了表型 MYC/BCL2 和微环境生物标志物以及遗传亚型在 DLBCL 预后中的独立和附加的预后效应，对于改善 DLBCL 分类和确定预后决定因素和治疗靶点非常重要。