The integration of single-cell RNA sequencing and spatial transcriptomics reveals the tumor microenvironment and spatial organization of testicular diffuse large B-cell lymphomas

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-...

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Main Authors: Xiaolong Wu, Jie Shi, Mujun Lu, Damin Yun, Sheng Gao, Longfei Hu, Fei Sun
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2025-07-01
Series:Genes and Diseases
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352304224002721
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author Xiaolong Wu
Jie Shi
Mujun Lu
Damin Yun
Sheng Gao
Longfei Hu
Fei Sun
author_facet Xiaolong Wu
Jie Shi
Mujun Lu
Damin Yun
Sheng Gao
Longfei Hu
Fei Sun
author_sort Xiaolong Wu
collection DOAJ
description 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.
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spelling doaj-art-b46bdb9df7ab4765a7fbcbdb8c570c3f2025-08-20T03:52:33ZengKeAi Communications Co., Ltd.Genes and Diseases2352-30422025-07-0112410147510.1016/j.gendis.2024.101475The integration of single-cell RNA sequencing and spatial transcriptomics reveals the tumor microenvironment and spatial organization of testicular diffuse large B-cell lymphomasXiaolong Wu0Jie Shi1Mujun Lu2Damin Yun3Sheng Gao4Longfei Hu5Fei Sun6Department of Urology and Andrology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310016, ChinaInstitute of Reproductive Medicine, Nantong University School of Medicine, Nantong, Jiangsu 226001, ChinaDepartment of Urology and Andrology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200001, ChinaInstitute of Reproductive Medicine, Nantong University School of Medicine, Nantong, Jiangsu 226001, ChinaInstitute of Reproductive Medicine, Nantong University School of Medicine, Nantong, Jiangsu 226001, ChinaSingleron Biotechnologies Ltd., 211 Pubin Road, Nanjing, Jiangsu 210061, ChinaDepartment of Urology and Andrology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310016, China; Corresponding author.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.http://www.sciencedirect.com/science/article/pii/S2352304224002721CREBE2FPrimary testicular diffuse large B-cell lymphoma (PT-DLBCL)Single-cell RNA-seqSpatial transcriptomics
spellingShingle Xiaolong Wu
Jie Shi
Mujun Lu
Damin Yun
Sheng Gao
Longfei Hu
Fei Sun
The integration of single-cell RNA sequencing and spatial transcriptomics reveals the tumor microenvironment and spatial organization of testicular diffuse large B-cell lymphomas
Genes and Diseases
CREB
E2F
Primary testicular diffuse large B-cell lymphoma (PT-DLBCL)
Single-cell RNA-seq
Spatial transcriptomics
title The integration of single-cell RNA sequencing and spatial transcriptomics reveals the tumor microenvironment and spatial organization of testicular diffuse large B-cell lymphomas
title_full The integration of single-cell RNA sequencing and spatial transcriptomics reveals the tumor microenvironment and spatial organization of testicular diffuse large B-cell lymphomas
title_fullStr The integration of single-cell RNA sequencing and spatial transcriptomics reveals the tumor microenvironment and spatial organization of testicular diffuse large B-cell lymphomas
title_full_unstemmed The integration of single-cell RNA sequencing and spatial transcriptomics reveals the tumor microenvironment and spatial organization of testicular diffuse large B-cell lymphomas
title_short The integration of single-cell RNA sequencing and spatial transcriptomics reveals the tumor microenvironment and spatial organization of testicular diffuse large B-cell lymphomas
title_sort integration of single cell rna sequencing and spatial transcriptomics reveals the tumor microenvironment and spatial organization of testicular diffuse large b cell lymphomas
topic CREB
E2F
Primary testicular diffuse large B-cell lymphoma (PT-DLBCL)
Single-cell RNA-seq
Spatial transcriptomics
url http://www.sciencedirect.com/science/article/pii/S2352304224002721
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