Targeted therapies prime oncogene-driven lung cancers for macrophage-mediated destruction
Kyle Vaccaro, … , Aaron N. Hata, Kipp Weiskopf
Kyle Vaccaro, … , Aaron N. Hata, Kipp Weiskopf
Published March 14, 2024
Citation Information: J Clin Invest. 2024;134(9):e169315. https://doi.org/10.1172/JCI169315.
View: Text | PDF
Research Article Oncology Article has an altmetric score of 30

Targeted therapies prime oncogene-driven lung cancers for macrophage-mediated destruction

Abstract

Macrophage immune checkpoint inhibitors, such as anti-CD47 antibodies, show promise in clinical trials for solid and hematologic malignancies. However, the best strategies to use these therapies remain unknown, and ongoing studies suggest they may be most effective when used in combination with other anticancer agents. Here, we developed an unbiased, high-throughput screening platform to identify drugs that render lung cancer cells more vulnerable to macrophage attack, and we found that therapeutic synergy exists between genotype-directed therapies and anti-CD47 antibodies. In validation studies, we found that the combination of genotype-directed therapies and CD47 blockade elicited robust phagocytosis and eliminated persister cells in vitro and maximized antitumor responses in vivo. Importantly, these findings broadly applied to lung cancers with various RTK/MAPK pathway alterations — including EGFR mutations, ALK fusions, or KRASG12C mutations. We observed downregulation of β2-microglobulin and CD73 as molecular mechanisms contributing to enhanced sensitivity to macrophage attack. Our findings demonstrate that dual inhibition of the RTK/MAPK pathway and the CD47/SIRPa axis is a promising immunotherapeutic strategy. Our study provides strong rationale for testing this therapeutic combination in patients with lung cancers bearing driver mutations.

Authors

Kyle Vaccaro, Juliet Allen, Troy W. Whitfield, Asaf Maoz, Sarah Reeves, José Velarde, Dian Yang, Anna Meglan, Juliano Ribeiro, Jasmine Blandin, Nicole Phan, George W. Bell, Aaron N. Hata, Kipp Weiskopf

×

Figure 4

Targeted inhibition of the MAPK pathway primes NSCLC cells for macrophage-mediated destruction.

Options: View larger image (or click on image) Download as PowerPoint
Targeted inhibition of the MAPK pathway primes NSCLC cells for macrophag...
(A) Growth of GFP+ NCI-H3122 cells (a human ALK+ NSCLC cell line) cocultured with primary human macrophages and treated with vehicle (PBS), an anti-CD47 antibody (10 μg/mL), and/or the indicated ALK-specific TKIs (1 μM). (B) Growth of GFP+ NCI-H3122 cells cocultured with macrophages with or without an anti-CD47 antibody (10 μg/mL) and varying concentrations of the ALK-specific TKI lorlatinib. IC50 of lorlatinib alone (PBS) = 10.29 nM (95% CI, 8.665–12.22) versus IC50 of lorlatinib+anti-CD47 = 2.135 nM (95% CI, 0.6934–6.261]). (C) Growth of GFP+ NCI-H358 cells (a human KRASG12C mutant NSCLC cell line) cocultured with macrophages and treated with vehicle (PBS), an anti-CD47 antibody (10 μg/mL), and/or the indicated KRASG12C inhibitors (1 μM). (D) Growth of GFP+ NCI-H358 cells cocultured with macrophages with or without an anti-CD47 antibody (10 μg/mL) and varying concentrations of the KRASG12C inhibitor sotorasib. IC50 of sotorasib alone (PBS) = 896.5 nM (95% CI, 558.6–1,697) versus IC50 of sotorasib+anti-CD47 = 10.30 nM (95% CI, 2.949–40.48). (E) Diagram depicting the EGFR/RAS/MAPK pathway. KRAS can signal via MAPK elements or the PI3K/AKT pathway. Specific inhibitors used in this study are indicated in red. Sotorasib is specific for KRASG12C. (F) Growth of GFP+ PC9 cells in coculture with macrophages with or without an anti-CD47 antibody (10 μg/mL) and varying EGFR/RAS/MAPK or PI3K/AKT pathway inhibitors. (G) Growth of GFP+ NCI-H358 cells in coculture with macrophages with or without an anti-CD47 antibody (10 μg/mL) and varying inhibitors, as in F. (A–G) Data represent individual cocultures with means (A, F, and G), mean ± SD on day 6.5 (B and D), or mean ± SEM (C) from 3 cocultures per donor using n = 4 independent macrophage donors. *P < 0.05, **P < 0.01, ***P < 0. 001, ****P < 0.0001 by 1-way ANOVA with Holm-Šidák multiple comparisons test.