Tumor Cells Modulate Macrophage Phenotype in a Novel In Vitro Co-Culture Model of the NSCLC Tumor Microenvironment

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

doi:10.1016/j.jtho.2022.06.011

Tumor Cells Modulate Macrophage Phenotype in a Novel In Vitro Co-Culture Model of the NSCLC Tumor Microenvironment

J Thorac Oncol. 2022 Oct;17(10):1178-1191. doi: 10.1016/j.jtho.2022.06.011. Epub 2022 Jul 5.

Authors

Josiah Voth Park¹, Raghav Chandra², Ling Cai³, Debolina Ganguly⁴, Huiyu Li⁵, Jason E Toombs², Luc Girard⁶, Rolf A Brekken⁷, John D Minna⁸

Affiliations

¹ Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas; Cancer Biology Graduate Program, University of Texas Southwestern Medical Center, Dallas, Texas.
² Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas; Division of Surgical Oncology, Department of Surgery, University of Texas Southwestern Medical Center, Dallas, Texas.
³ Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas; Quantitative Biomedical Research Center, Department of Population and Data Science, University of Texas Southwestern Medical Center, Dallas, Texas; Children's Research Institute, University of Texas Southwestern Medical Center, Dallas, Texas.
⁴ Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas; Cancer Biology Graduate Program, University of Texas Southwestern Medical Center, Dallas, Texas; Division of Surgical Oncology, Department of Surgery, University of Texas Southwestern Medical Center, Dallas, Texas.
⁵ Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas; Children's Research Institute, University of Texas Southwestern Medical Center, Dallas, Texas.
⁶ Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas; Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas; Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas.
⁷ Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas; Cancer Biology Graduate Program, University of Texas Southwestern Medical Center, Dallas, Texas; Division of Surgical Oncology, Department of Surgery, University of Texas Southwestern Medical Center, Dallas, Texas; Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas; Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas.
⁸ Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas; Cancer Biology Graduate Program, University of Texas Southwestern Medical Center, Dallas, Texas; Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas; Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas; Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas. Electronic address: John.Minna@utsouthwestern.edu.

Abstract

Introduction: Macrophage phenotype in the tumor microenvironment correlates with prognosis in NSCLC. Immunosuppressive macrophages promote tumor progression, whereas proinflammatory macrophages may drive an antitumor immune response. How individual NSCLCs affect macrophage phenotype is a major knowledge gap.

Methods: To systematically study the impact of lung cancer cells on macrophage phenotypes, we developed an in vitro co-culture model that consisted of molecularly and clinically annotated patient-derived NSCLC lines, human cancer-associated fibroblasts, and murine macrophages. Induced macrophage phenotype was studied through quantitative real-time polymerase chain reaction and validated in vivo using NSCLC xenografts through quantitative immunohistochemistry and clinically with The Cancer Genome Atlas (TCGA)-"matched" patient tumors.

Results: A total of 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 affect M2:M1 ratios in matched TCGA samples. In vivo, xenograft tumors established from high Arginase-1-inducing lines (Arg^hi) had a significantly elevated density of Arg1+ macrophages. Matched TCGA clinical samples to Arg^hi NSCLC lines had a significantly higher ratio of M2:M1 macrophages (p = 0.0361).

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

Keywords: In vitro co-culture model; Macrophage phenotype; Non–small cell lung cancer; Tumor microenvironment.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Animals
Arginase / genetics
Arginase / metabolism
Arginase / therapeutic use
Carcinoma, Non-Small-Cell Lung* / drug therapy
Coculture Techniques
Humans
Lung Neoplasms* / drug therapy
Macrophages / metabolism
Mice
Phenotype
Tumor Microenvironment

Substances

Arginase

Abstract

Publication types

MeSH terms

Substances

Grants and funding