Moody Sarah C, Fietz Daniela, Nathaniel Benedict, Frydenberg Mark, Tran Ben, Schuppe Hans-Christian, Loveland Kate L
Centre for Reproductive Health, Hudson Institute of Medical Research, Clayton, Victoria, Australia.
Department of Molecular and Translational Sciences, School of Clinical Sciences, Monash University, Clayton, Victoria, Australia.
Andrology. 2025 Jul 22. doi: 10.1111/andr.70100.
Testicular germ cell tumours (TGCTs) are amongst the most common malignancies in young men, and their incidence is increasing worldwide. Tissue heterogeneity hampers efforts to understand how TGCT precursors (termed germ cell neoplasia in situ; GCNIS) emerge and progress, restricting elucidation of new strategies for diagnosis and management.
This study reports the use of spatial transcriptomic analysis in TGCT tissue sections.
Over 90 regions of interest (ROIs) were identified in sections from four TGCT patients' samples, three with non-seminoma, one seminoma. Transcripts in each ROI were sequenced and examined using the NanoString GeoMx spatial whole transcriptomics workflow, the values normalised and analysed using Degust RNA-Seq and Ingenuity Pathway Analysis software.
The distribution and expression of functional markers in specific cell types was used to map individual tumours, GCNIS, and tumour-adjacent regions. Significant heterogeneity in TGCTs and surrounding areas is documented between patients and across different regions in the same tumour. Immune cell-related transcripts encoding macrophage, T cell, B cell, natural killer cell, dendritic cells and neutrophil subsets were identified as contributing substantially to tumour heterogeneity. Assessment of ROIs containing GCNIS and areas immediately adjacent from two individual non-seminoma tumour samples identified the TGFβ family as contributing to upstream regulation of transcripts in both patients; known activin A target genes were differentially expressed between the GCNIS and microenvironment ROIs. In addition, two discrete tumour areas within the seminoma sample displayed distinct transcript profiles, one featuring higher levels of immune cell-related transcripts, and the other TGFβ superfamily transcripts.
These findings highlight aspects of the complex challenge faced while seeking therapeutic targets to enable tumour spread restriction. However, these outcomes reinforce knowledge that TGFβ family members can influence seminoma fate and provide new evidence of their potential contribution to the transition of GCNIS cells into tumours.
睾丸生殖细胞肿瘤（TGCTs）是年轻男性中最常见的恶性肿瘤之一，且其发病率在全球范围内呈上升趋势。组织异质性阻碍了人们对TGCT前体（原位生殖细胞肿瘤；GCNIS）如何发生和发展的理解，限制了新的诊断和管理策略的阐明。
本研究报告了空间转录组分析在TGCT组织切片中的应用。
在来自4例TGCT患者样本的切片中识别出90多个感兴趣区域（ROIs），其中3例为非精原细胞瘤，1例为精原细胞瘤。使用NanoString GeoMx空间全转录组学工作流程对每个ROI中的转录本进行测序和检测，使用Degust RNA-Seq和Ingenuity Pathway Analysis软件对值进行归一化和分析。
特定细胞类型中功能标记的分布和表达被用于绘制个体肿瘤、GCNIS和肿瘤相邻区域的图谱。记录了患者之间以及同一肿瘤不同区域之间TGCTs及其周围区域的显著异质性。编码巨噬细胞、T细胞、B细胞、自然杀伤细胞、树突状细胞和中性粒细胞亚群的免疫细胞相关转录本被确定为对肿瘤异质性有显著贡献。对来自两个个体非精原细胞瘤肿瘤样本中包含GCNIS的ROIs和紧邻区域的评估确定，TGFβ家族在两名患者中均对转录本的上游调控有贡献；已知的激活素A靶基因在GCNIS和微环境ROIs之间差异表达。此外，精原细胞瘤样本中的两个离散肿瘤区域显示出不同的转录谱，一个具有较高水平的免疫细胞相关转录本，另一个具有TGFβ超家族转录本。
这些发现突出了在寻找限制肿瘤扩散的治疗靶点时所面临的复杂挑战的一些方面。然而，这些结果强化了TGFβ家族成员可影响精原细胞瘤命运的认识，并为它们对GCNIS细胞向肿瘤转变的潜在贡献提供了新证据。
