Background/Objectives: Penile squamous cell carcinoma (PSCC) is a rare malignancy with poor prognosis in advanced and recurrent disease, and therapeutic options remain limited. Increasing evidence suggests that the tumor immune microenvironment (TIME), including immune cell composition and spatial organization, plays a critical role in tumor progression and survival outcomes. This study aimed to characterize immune cell density and geospatial clustering patterns within the TIME of PSCC and to evaluate their associations with clinical outcomes. Methods: Multiplex immunofluorescence (mIF) was performed on tumor samples from 57 patients with PSCC using a panel of immune markers to identify lymphoid and myeloid cell populations. Immune cell densities were quantified within tumoral and stromal compartments. Spatial relationships among immune cells and between immune cells and tumor cells were analyzed using point pattern analysis. Survival outcomes, including overall survival (OS), recurrence-free survival (RFS), and cancer-specific survival (CSS), were assessed using Kaplan–Meier methods and Cox proportional hazards models, with analyses stratified by nodal and human papillomavirus (HPV) status. Results: Higher intratumoral and stromal densities of pro-immunogenic M1 macrophages were associated with improved OS. Increased densities of CD3+CD4+helper T cells in both compartments were also associated with favorable survival outcomes. In contrast, close clustering of pro-tumorigenic M2 macrophages with tumor cells and with one another was associated with worse OS, RFS, and CSS. Bivariate clustering of helper T cells with tumor cells was associated with improved OS, including among patients with node-positive disease. Survival outcomes did not differ significantly by HPV status in patients with high helper T cell clustering. Conclusions: Immune cell density and spatial organization within the TIME are associated with survival outcomes in PSCC. Favorable patterns involving helper T cells and M1 macrophages correlate with improved survival, whereas clustering of M2 macrophages is associated with poorer outcomes, supporting the relevance of spatial immune profiling in this disease.
背景/目的：阴茎鳞状细胞癌（PSCC）是一种罕见恶性肿瘤，在晚期和复发性疾病中预后较差，且治疗选择有限。越来越多的证据表明，肿瘤免疫微环境（TIME），包括免疫细胞组成和空间结构，在肿瘤进展和生存结局中起着关键作用。本研究旨在描述PSCC肿瘤免疫微环境中的免疫细胞密度和地理空间聚类模式，并评估其与临床结局的关联。方法：使用一组免疫标志物对57例PSCC患者的肿瘤样本进行多重免疫荧光（mIF）检测，以识别淋巴和髓系细胞群。量化肿瘤区域和基质区域内的免疫细胞密度。采用点模式分析评估免疫细胞之间以及免疫细胞与肿瘤细胞之间的空间关系。使用Kaplan-Meier法和Cox比例风险模型评估总生存期（OS）、无复发生存期（RFS）和癌症特异性生存期（CSS）等生存结局，分析按淋巴结状态和人乳头瘤病毒（HPV）状态进行分层。结果：肿瘤内和基质中促免疫原性M1巨噬细胞密度较高与OS改善相关。两个区域中CD3+CD4+辅助性T细胞密度增加也与较好的生存结局相关。相反，促肿瘤发生M2巨噬细胞与肿瘤细胞及彼此之间的紧密聚类与较差的OS、RFS和CSS相关。辅助性T细胞与肿瘤细胞的双变量聚类与OS改善相关，在淋巴结阳性患者中亦如此。在辅助性T细胞高聚类患者中，不同HPV状态的生存结局无显著差异。结论：肿瘤免疫微环境中的免疫细胞密度和空间结构与PSCC的生存结局相关。涉及辅助性T细胞和M1巨噬细胞的有利模式与生存改善相关，而M2巨噬细胞的聚类则与较差结局相关，这支持了空间免疫谱分析在该疾病中的相关性。
肿瘤(癌症)患者之家