Lopez Diana C, Robbins Yvette L, Kowalczyk Joshua T, Lassoued Wiem, Gulley James L, Miettinen Markku M, Gallia Gary L, Allen Clint T, Hodge James W, London Nyall R
Sinonasal and Skull Base Tumor Program, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, MD, United States.
Cleveland Clinic Lerner College of Medicine, Cleveland Clinic, Cleveland, OH, United States.
Front Oncol. 2022 Oct 21;12:1012058. doi: 10.3389/fonc.2022.1012058. eCollection 2022.
Chordoma is a rare, invasive, and devastating bone malignancy of residual notochord tissue that arises at the skull base, sacrum, or spine. In order to maximize immunotherapeutic approaches as a potential treatment strategy in chordoma it is important to fully characterize the tumor immune microenvironment (TIME). Multispectral immunofluorescence (MIF) allows for comprehensive evaluation of tumor compartments, molecular co-expression, and immune cell spatial relationships. Here we implement MIF to define the myeloid, T cell, and natural killer (NK) cell compartments in an effort to guide rational design of immunotherapeutic strategies for chordoma.
Chordoma tumor tissue from 57 patients was evaluated using MIF. Three panels were validated to assess myeloid cell, T cell, and NK cell populations. Slides were stained using an automated system and HALO software objective analysis was utilized for quantitative immune cell density and spatial comparisons between tumor and stroma compartments.
Chordoma TIME analysis revealed macrophage infiltration of the tumor parenchyma at a significantly higher density than stroma. In contrast, helper T cells, cytotoxic T cells, and T regulatory cells were significantly more abundant in stroma versus tumor. T cell compartment infiltration more commonly demonstrated a tumor parenchymal exclusion pattern, most markedly among cytotoxic T cells. NK cells were sparsely found within the chordoma TIME and few were in an activated state. No immune composition differences were seen in chordomas originating from diverse anatomic sites or between those resected at primary versus advanced disease stage.
This is the first comprehensive evaluation of the chordoma TIME including myeloid, T cell, and NK cell appraisal using MIF. Our findings demonstrate that myeloid cells significantly infiltrate chordoma tumor parenchyma while T cells tend to be tumor parenchymal excluded with high stromal infiltration. On average, myeloid cells are found nearer to target tumor cells than T cells, potentially resulting in restriction of T effector cell function. This study suggests that future immunotherapy combinations for chordoma should be aimed at decreasing myeloid cell suppressive function while enhancing cytotoxic T cell and NK cell killing.
脊索瘤是一种罕见的、侵袭性的、毁灭性的骨恶性肿瘤，起源于残留的脊索组织，好发于颅底、骶骨或脊柱。为了将免疫治疗方法作为脊索瘤的一种潜在治疗策略最大化，充分表征肿瘤免疫微环境（TIME）很重要。多光谱免疫荧光（MIF）能够全面评估肿瘤区域、分子共表达以及免疫细胞的空间关系。在此，我们应用MIF来定义髓系细胞、T细胞和自然杀伤（NK）细胞区域，以指导脊索瘤免疫治疗策略的合理设计。
使用MIF对57例患者的脊索瘤肿瘤组织进行评估。验证了三个检测板以评估髓系细胞、T细胞和NK细胞群体。玻片采用自动系统进行染色，并利用HALO软件进行客观分析，以定量免疫细胞密度，并比较肿瘤和基质区域之间的空间差异。
脊索瘤TIME分析显示，肿瘤实质内巨噬细胞浸润密度显著高于基质。相比之下，辅助性T细胞、细胞毒性T细胞和调节性T细胞在基质中比在肿瘤中明显更丰富。T细胞区域浸润更常见地表现为肿瘤实质排斥模式，在细胞毒性T细胞中最为明显。在脊索瘤TIME中NK细胞稀少，且很少处于激活状态。源自不同解剖部位的脊索瘤或在原发性疾病与晚期疾病阶段切除的脊索瘤之间，未见免疫组成差异。
这是首次使用MIF对脊索瘤TIME进行全面评估，包括髓系细胞、T细胞和NK细胞评估。我们的研究结果表明，髓系细胞显著浸润脊索瘤肿瘤实质，而T细胞倾向于被肿瘤实质排斥，但在基质中有高度浸润。平均而言，髓系细胞比T细胞更靠近肿瘤靶细胞，这可能导致T效应细胞功能受限。这项研究表明，未来脊索瘤的免疫治疗联合方案应旨在降低髓系细胞的抑制功能，同时增强细胞毒性T细胞和NK细胞的杀伤作用。
