A new digital droplet PCR method for looking at epigenetics in diffuse large B‐cell lymphomas: The role of BMI1, EZH2, and USP22 genes

IF 2.2 4区医学 Q3 HEMATOLOGY

International Journal of Laboratory Hematology Pub Date : 2024-09-10 DOI:10.1111/ijlh.14363

Alessio Lusci Gemignani, Robel Papotti, Riccardo Bomben, Valter Gattei, Samantha Pozzi, Valentina Donati, Stefania Bettelli, Elisa Forti, Giovanna Mansueto, Arianna Di Napoli, Maria Christina Cox, Leonardo Flenghi, Pietro Rossi, Guido Volpe, Dimitri Dardanis, Clara Bono, Francesca Guerrini, Riccardo Morganti, Stefano Sacchi, Sara Galimberti

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引用次数: 0

Abstract

IntroductionEpigenetics has been shown to be relevant in oncology: BMI1 overexpression has been reported in leukemias, EZH2 mutations have been found in follicular lymphoma, and USP22 seems to stabilize BMI1 protein. In this study, we measured the expression of BMI1, EZH2, and USP22 in lymph nodes from 56 diffuse large B‐cell lymphoma (DLBCL) patients.MethodsA new multiplex digital droplet PCR (ddPCR) has been set up to measure the expression of 4 genes (BMI1, EZH2, USP22, and GAPDH) in the same reaction on RNA extracted from paraffin‐embedded tissues.ResultsThe specificity of ddPCR was confirmed by a 100% alignment on the BLAST platform and its repeatability demonstrated by duplicates. A strict correlation between expression of BMI1 and EZH2 and BMI1 and USP22 has been found, and high expression of these genes was correlated with extra‐nodal lymphomas. Progression‐free survival (PFS) and overall survival (OS) were conditioned by IPI, bone marrow infiltration, and the complete response achievement. High levels of BMI1 and USP22 did not condition the response to therapy, but impaired the PFS, especially for patients defined at “high risk” based on the cell of origin (no germinal center [GCB]), high BCL2 expression, and IPI 3‐5. In this subgroup, the probability of relapse/progression was twice higher than that of patients carrying low BMI1 and USP22 levels.ConclusionHigh expression of BMI1 and of USP22 might be a poor prognostic factor in DLBCL, and might represent the target for novel inhibitors.

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研究弥漫大 B 细胞淋巴瘤表观遗传学的新型数字液滴 PCR 方法：BMI1、EZH2 和 USP22 基因的作用

导言表观遗传学已被证明与肿瘤学有关：据报道，BMI1在白血病中过表达，EZH2突变在滤泡性淋巴瘤中被发现，而USP22似乎能稳定BMI1蛋白。本研究测定了 56 例弥漫大 B 细胞淋巴瘤（DLBCL）患者淋巴结中 BMI1、EZH2 和 USP22 的表达。方法建立了一种新的多重数字液滴 PCR（ddPCR），在同一反应中测量从石蜡包埋组织中提取的 RNA 中 4 个基因（BMI1、EZH2、USP22 和 GAPDH）的表达。结果发现，BMI1 和 EZH2 以及 BMI1 和 USP22 的表达之间存在严格的相关性，这些基因的高表达与结外淋巴瘤相关。无进展生存期（PFS）和总生存期（OS）受 IPI、骨髓浸润和完全反应成就的影响。高水平的BMI1和USP22不会影响治疗反应，但会影响无进展生存期，尤其是根据起源细胞（无生殖中心[GCB]）、BCL2高表达和IPI 3-5定义的 "高危 "患者。结论BMI1和USP22的高表达可能是DLBCL的不良预后因素，可能是新型抑制剂的靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International Journal of Laboratory Hematology 医学-血液学

CiteScore

4.50

自引率

6.70%

发文量

211

审稿时长

6-12 weeks

期刊介绍： The International Journal of Laboratory Hematology provides a forum for the communication of new developments, research topics and the practice of laboratory haematology. The journal publishes invited reviews, full length original articles, and correspondence. The International Journal of Laboratory Hematology is the official journal of the International Society for Laboratory Hematology, which addresses the following sub-disciplines: cellular analysis, flow cytometry, haemostasis and thrombosis, molecular diagnostics, haematology informatics, haemoglobinopathies, point of care testing, standards and guidelines. The journal was launched in 2006 as the successor to Clinical and Laboratory Hematology, which was first published in 1979. An active and positive editorial policy ensures that work of a high scientific standard is reported, in order to bridge the gap between practical and academic aspects of laboratory haematology.