Katey S. S. Enfield, Spencer D. Martin, Sonia H Y Kung, P. Gallagher, C. MacAulay, M. Guillaud, W. Lam
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引用次数: 0
Abstract
Immune cells are a major component of the tumor microenvironment (TME). The spatial organization of immune cell subpopulations within the TME is recognized to have biologic significance and clinical relevance. For example, spatial organization of immune cell subsets within the TME is critical for the inhibition of cytotoxic T-cell activity through direct interaction of ligand (PD-L1) with receptor (PD-1)). However, precise spatial deconvolution is limited by the lack of imaging algorithms for in situ multiplex single cell analyses as flow cytometry does not preserve data in the spatial dimension. To this end, we have developed a hyperspectral imaging platform designed for analyzing multichannel immunohistochemical-stained tissue sections for generating cell density data and reconstructing spatial architecture for tumor biology as well as clinical association studies. Whole-tissue sections from 20 lung adenocarcinomas with at least 5 years’ follow-up were stained for CD3 (pan-T cell), CD8 (cytotoxic T cell), and CD79a (B cell and plasma cell) and counterstained with hematoxylin. Multispectral images were acquired for five fields of view and analyzed to quantify cell types. Regions of Interest (ROIs) were then identified and analyzed in order to quantify cell-cell spatial relationships. Nonrandom patterns of immune cell distributions were identified using the Monte Carlo resampling method (500 iterations). Cell counts, densities, spatial relationships, and significant immune cell distributions were associated with clinical features (Kruskal - Wallis p Our analysis generated 234 image files for analysis, with an average of 16,400 cells per image. The densities of intratumoral CD8+ cytotoxic T cells were significantly higher in nonrecurrent cases, agreeing with literature reports. Similarly, cell sociology deductions identified relationships associated with metastasis: tumor cells in nonmetastatic cases had increased numbers of CD8+ cytotoxic T-cell neighbors. Following Monte Carlo analysis, nonrandom cell~cell spatial proximities emerged that were not identified at a cell density level. We have developed a hyperspectral imaging platform capable of quantifying cell-cell spatial relationships within tissue sections. This technology can be applied to larger clinical cohorts for the study of therapeutically targetable immune cell subsets with the goal of identifying patterns that correlate with clinical response and patient outcome. This abstract is also being presented as Poster B16. Citation Format: Katey S.S. Enfield, Spencer D. Martin, Sonia H.Y. Kung, Paul Gallagher, Calum E. MacAulay, Martial Guillaud, Wan L. Lam. Hyperspectral imaging tools capture the spatial organization of cell subsets within the tumor microenvironment [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr PR11.
免疫细胞是肿瘤微环境(TME)的主要组成部分。TME内免疫细胞亚群的空间组织被认为具有生物学意义和临床相关性。例如,TME内免疫细胞亚群的空间组织对于通过配体(PD-L1)与受体(PD-1)的直接相互作用抑制细胞毒性t细胞活性至关重要。然而,精确的空间反褶积由于缺乏原位多重单细胞分析的成像算法而受到限制,因为流式细胞术不能在空间维度上保存数据。为此,我们开发了一个高光谱成像平台,用于分析多通道免疫组织化学染色的组织切片,以生成细胞密度数据和重建肿瘤生物学和临床关联研究的空间结构。对20例肺腺癌进行至少5年随访的全组织切片进行CD3(泛T细胞)、CD8(细胞毒性T细胞)和CD79a (B细胞和浆细胞)染色,并用苏木精反染。获得5个视场的多光谱图像,并对其进行分析以量化细胞类型。然后识别和分析感兴趣区域(roi),以量化细胞-细胞空间关系。利用蒙特卡罗重采样法(500次迭代)识别免疫细胞分布的非随机模式。细胞计数、密度、空间关系和显著的免疫细胞分布与临床特征相关(Kruskal - Wallis p)。我们的分析生成了234个图像文件用于分析,平均每张图像有16,400个细胞。非复发病例的瘤内CD8+细胞毒性T细胞密度显著升高,与文献报道一致。类似地,细胞社会学推断确定了与转移相关的关系:非转移病例的肿瘤细胞CD8+细胞毒性t细胞邻居数量增加。在蒙特卡罗分析之后,出现了在细胞密度水平上无法识别的非随机细胞~细胞空间邻近性。我们开发了一种高光谱成像平台,能够定量组织切片内的细胞-细胞空间关系。这项技术可以应用于更大的临床队列,用于研究治疗上可靶向的免疫细胞亚群,目的是确定与临床反应和患者结果相关的模式。此摘要也以海报B16的形式呈现。引文格式:Katey S.S. Enfield, Spencer D. Martin, Sonia H.Y. Kung, Paul Gallagher, Calum E. MacAulay, Martial Guillaud, Wan L. Lam。高光谱成像工具捕获肿瘤微环境中细胞亚群的空间组织[摘要]。第五届AACR-IASLC国际联合会议论文集:肺癌转化科学从实验室到临床;2018年1月8日至11日;费城(PA): AACR;临床癌症杂志,2018;24(17):摘要nr PR11。