非小细胞肺癌肿瘤微环境体外共培养模型中肿瘤细胞调节巨噬细胞表型

SCI

23 July 2022

Tumor cells modulate macrophage phenotype in a novel in vitro co-culture model of the non-small cell lung cancer tumor microenvironment

(Journal of Thoracic Oncology, 20.121)

Josiah Voth Park, Raghav Chandra, Ling Cai, Debolina Ganguly, Huiyu Li, Jason E.Toombs, Luc Girard, Rolf A. Brekken, John D. Minna

CORRESPONDENCE TO: John.Minna@utsouthwestern.edu

Introduction 引言

Macrophage phenotype in the tumor microenvironment correlates with prognosis in non-small cell lung cancer (NSCLC). Immunosuppressive macrophages promote tumor progression, while pro-inflammatory macrophages may drive an anti-tumor immune response. How individual NSCLCs impact macrophage phenotype is a major knowledge gap.

肿瘤微环境中的巨噬细胞表型与非小细胞肺癌（NSCLC）的预后相关。免疫抑制性巨噬细胞促进肿瘤进展，而促炎性巨噬细胞可能驱动抗肿瘤免疫反应。个体非小细胞肺癌如何影响巨噬细胞表型是一个主要的知识缺口。

Methods 方法

To systematically study the impact of lung cancer cells on macrophage phenotypes, we developed an in vitro co-culture model comprised of molecularly and clinically-annotated patient-derived NSCLC lines, human cancer-associated fibroblasts, and murine macrophages. Induced macrophage phenotype was studied through qRT-PCR and validated in vivo using NSCLC xenografts through quantitative immunohistochemistry and clinically with TCGA “matched” patient tumors.

为了系统地研究肺癌细胞对巨噬细胞表型的影响，我们开发了一种体外共培养模型，该模型由分子和临床注释的患者来源的非小细胞肺癌细胞系、人肿瘤相关成纤维细胞和小鼠巨噬细胞组成。通过qRT-PCR研究诱导的巨噬细胞表型，并通过定量免疫组织化学在体内使用非小细胞肺癌异种移植物进行验证，并与TCGA“匹配”患者肿瘤进行临床验证。

Results 结果

72 NSCLC cell lines were studied. The most frequent highly induced macrophage-related gene was Arginase-1, reflecting an immunosuppressive M2-like phenotype. This was independent of multiple clinicopathologic factors, which also did not impact M2:M1 ratios in matched TCGA samples. In vivo, xenograft tumors established from high Arginase-1-inducing lines (Arghi) had a significantly elevated density of Arg1+ macrophages. Matched TCGA clinical samples to Arghi NSCLC lines had a significantly higher ratio of M2:M1 macrophages. 

研究了72株非小细胞肺癌细胞系。最常见的高诱导巨噬细胞相关基因是精氨酸酶-1，反映免疫抑制M2样表型。这与多种临床病理因素无关，也不影响匹配TCGA样本中M2:M1的比率。在体内，由高精氨酸酶-1诱导系（Arghi）建立的异种移植瘤具有显著升高的Arg1+巨噬细胞密度。与Arghi非小细胞肺癌细胞系匹配的TCGA临床样本具有显著更高的M2:M1巨噬细胞比率。

Conclusions 结论

In our in vitro co-culture model, a large panel of patient-derived NSCLC lines most frequently induced high expression Arginase-1in co-cultured mouse macrophages, independent of major clinicopathologic and oncogenotype-related factors. Arghi cluster-matched TCGA tumors contained a higher ratio of M2:M1 macrophages. Thus, this in vitro model reproducibly characterizes how individual NSCLCs modulate macrophage phenotype, correlates with macrophage polarization in clinical samples, and can serve as an accessible platform for further investigation of macrophage-specific therapeutic strategies.

在我们的体外共培养模型中，大量患者来源的非小细胞肺癌细胞系最常在共培养的小鼠巨噬细胞中诱导高表达精氨酸酶-1，与主要临床病理学和癌基因型相关因素无关。Arghi簇匹配的TCGA肿瘤含有较高的M2:M1巨噬细胞比率。因此，该体外模型可重复性地描述单个非小细胞肺癌如何调节巨噬细胞表型，与临床样本中的巨噬细胞极化相关，并可作为进一步研究巨噬细胞特异性治疗策略的可用平台。