Yeung Tony, Zhang Yi, Zhou Qianghua, Burack Richard
Pathology and Laboratory Medicine, University of Rochester Medical Center, 601 Elmwood Avenue, Rochester, NY 14642, USA.
Department of Laboratory Medicine and Pathobiology, University of Toronto, 1 King's College Circle, 6 Floor, Toronto M5S 1A8, ON, Canada.
J Pathol Inform. 2025 Aug 6;19:100465. doi: 10.1016/j.jpi.2025.100465. eCollection 2025 Nov.
Evaluation of tumor infiltrating lymphocytes as recommended by current guidelines is largely based on stromal regions within the tumor. In the context of epithelial malignancies, the epithelial region and the epithelial-stromal interface are not assessed, because of technical difficulties in manually discerning lymphocytes when admixed with epithelial tumor cells. The inability to quantify immune cells in epithelial-associated areas may negatively impact evaluation of patient response to immune checkpoint therapies. Innovative spatial analysis techniques have emerged that can directly address challenges associated with quantification of lymphocytes in specialized regions like the interface. In this study, we apply supervised neighborhood clustering analysis (via an open-source application CytoMAP) to assess the spatial distribution of CD8+ T cells, CD8+ TIM3+ (T cell immunoglobulin and mucin-domain containing-3) exhausted T cells, and TIM3+ CD8- macrophages on a gynecological tumor microarray. Neighborhood clustering analysis is adept at objectively mapping the epithelial-stromal interface alongside the epithelial and stromal region of each tumor under a three-compartment model. When tumors are partitioned by the conventional two-compartment model (epithelial and stromal region only), the highest density of total CD8+ T cells is found in the stromal region in a slight majority of tumors. In contrast, the interface region surpasses both the epithelial and stromal region in holding the highest density of CD8+ T cells when this unique region is incorporated into the three-compartment model. Further subset analysis shows higher proportion of CD8+ TIM3+ exhausted T cells within the interface and epithelial region, as compared to CD8+ TIM3- T cells which span from the stroma to the interface. These results highlight the utility of implementing quantitative spatial technique and immune subset analysis in the assessment of tumor infiltrating lymphocytes, and underscore the potential significance of the under-reported tumor epithelial-stromal interface.
按照当前指南的建议，对肿瘤浸润淋巴细胞的评估主要基于肿瘤内的基质区域。在上皮性恶性肿瘤的情况下，上皮区域和上皮-基质界面未被评估，因为当淋巴细胞与上皮肿瘤细胞混合时，手动辨别淋巴细胞存在技术困难。无法对上皮相关区域的免疫细胞进行量化可能会对患者对免疫检查点疗法的反应评估产生负面影响。已经出现了创新的空间分析技术，这些技术可以直接应对与在界面等特殊区域量化淋巴细胞相关的挑战。在本研究中，我们应用监督邻域聚类分析（通过开源应用程序CytoMAP）来评估妇科肿瘤微阵列上CD8 + T细胞、CD8 + TIM3 +（含T细胞免疫球蛋白和粘蛋白结构域3）耗竭性T细胞以及TIM3 + CD8 -巨噬细胞的空间分布。邻域聚类分析擅长在三室模型下客观地绘制每个肿瘤的上皮和基质区域以及上皮-基质界面。当肿瘤按照传统的两室模型（仅上皮和基质区域）划分时，在大多数肿瘤中，总CD8 + T细胞的最高密度出现在基质区域。相比之下，当将这个独特区域纳入三室模型时，界面区域在CD8 + T细胞密度最高方面超过了上皮和基质区域。进一步的亚组分析表明，与从基质延伸到界面的CD8 + TIM3 - T细胞相比，界面区域和上皮区域内CD8 + TIM3 +耗竭性T细胞的比例更高。这些结果突出了在评估肿瘤浸润淋巴细胞时实施定量空间技术和免疫亚组分析的实用性，并强调了报告不足的肿瘤上皮-基质界面的潜在重要性。
