Wu Xiaolong, Shi Jie, Lu Mujun, Yun Damin, Gao Sheng, Hu Longfei, Sun Fei
Department of Urology and Andrology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310016, China.
Institute of Reproductive Medicine, Nantong University School of Medicine, Nantong, Jiangsu 226001, China.
Genes Dis. 2024 Nov 30;12(4):101475. doi: 10.1016/j.gendis.2024.101475. eCollection 2025 Jul.
Primary testicular diffuse large B-cell lymphomas (PT-DLBCL) are a collection of 1%-9% of testicular tumors. However, the characterization of the tumor microenvironment and spatial organization of PT-DLBCL is poorly understood. We profiled the transcriptomes of 19,559 single cells derived from a PT-DLBCL patient via single-cell RNA sequencing. We found that the tumor microenvironment was majorly composed of three exhausted CD8 T cell subpopulations and two B cell subpopulations, and the genetic heterogeneity was further analyzed. Then, transcription factors related to PT-DLBCL cell proliferation and development were identified. Our results demonstrated that inhibiting E2F and CREB could decrease cell proliferation, induce apoptosis in human B-lymphoma cells, and inhibit tumor growth in xenograft testicular DLBCL models. Subsequently, chromatin immunoprecipitation sequencing was performed to identify the enriched loci of E2F and CREB that regulate human B-lymphoma cell proliferation and apoptosis. To annotate the precise spatial cellular composition of testicular DLBCL, we performed spatial transcriptomics. The spatial organization of PT-DLBCL, especially the spatial location of exhausted CD8 T and B cells, was identified. Concurrently, we delineated the expression patterns of key genes, including MALAT1, RPS3A, RPS7, RPS23, RPS27A, IGHM, HINT1, and HSPA8, across various regions. In this study, we unveiled the spatial architecture of the tumor microenvironment in DLBCL, where exhausted T cells were strategically positioned around tumor B cells, and macrophages, in turn, encircled the exhausted T cells. Inhibition of E2F and CREB in the tumor microenvironment may be a novel therapeutic option for testicular DLBCL patients.
原发性睾丸弥漫性大B细胞淋巴瘤（PT-DLBCL）占睾丸肿瘤的1%-9%。然而，人们对PT-DLBCL的肿瘤微环境特征和空间组织了解甚少。我们通过单细胞RNA测序对一名PT-DLBCL患者的19559个单细胞的转录组进行了分析。我们发现肿瘤微环境主要由三个耗竭的CD8 T细胞亚群和两个B细胞亚群组成，并进一步分析了基因异质性。然后，鉴定了与PT-DLBCL细胞增殖和发育相关的转录因子。我们的结果表明，抑制E2F和CREB可降低细胞增殖，诱导人B淋巴瘤细胞凋亡，并抑制异种移植睾丸DLBCL模型中的肿瘤生长。随后，进行了染色质免疫沉淀测序，以确定调节人B淋巴瘤细胞增殖和凋亡的E2F和CREB的富集位点。为了注释睾丸DLBCL精确的空间细胞组成，我们进行了空间转录组学分析。确定了PT-DLBCL的空间组织，特别是耗竭的CD8 T细胞和B细胞的空间位置。同时，我们描绘了关键基因，包括MALAT1、RPS3A、RPS7、RPS23、RPS27A、IGHM、HINT1和HSPA8在不同区域中的表达模式。在本研究中，我们揭示了DLBCL肿瘤微环境的空间结构，其中耗竭的T细胞战略性地定位在肿瘤B细胞周围，而巨噬细胞又环绕着耗竭的T细胞。抑制肿瘤微环境中的E2F和CREB可能是睾丸DLBCL患者的一种新的治疗选择。
