Reprogramming of PD-1+ M2-like tumor-associated macrophages with anti–PD-L1 and lenalidomide in cutaneous T cell lymphoma
Zhen Han, Xiwei Wu, Hanjun Qin, Yate-Ching Yuan, Daniel Schmolze, Chingyu Su, Jasmine Zain, Lilach Moyal, Emmilia Hodak, James F. Sanchez, Peter P. Lee, Mingye Feng, Steven T. Rosen, Christiane Querfeld
Zhen Han, Xiwei Wu, Hanjun Qin, Yate-Ching Yuan, Daniel Schmolze, Chingyu Su, Jasmine Zain, Lilach Moyal, Emmilia Hodak, James F. Sanchez, Peter P. Lee, Mingye Feng, Steven T. Rosen, Christiane Querfeld
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Research Article Dermatology

Reprogramming of PD-1+ M2-like tumor-associated macrophages with anti–PD-L1 and lenalidomide in cutaneous T cell lymphoma

Abstract

Cutaneous T cell lymphoma (CTCL) is a disfiguring and incurable disease characterized by skin-homing malignant T cells surrounded by immune cells that promote CTCL growth through an immunosuppressive tumor microenvironment (TME). Preliminary data from our phase I clinical trial of anti–programmed cell death ligand 1 (anti–PD-L1) combined with lenalidomide in patients with relapsed/refractory CTCL demonstrated promising clinical efficacy. In the current study, we analyzed the CTCL TME, which revealed a predominant PD-1+ M2-like tumor-associated macrophage (TAM) subtype with upregulated NF-κB and JAK/STAT signaling pathways and an aberrant cytokine and chemokine profile. Our in vitro studies investigated the effects of anti–PD-L1 and lenalidomide on PD-1+ M2-like TAMs. The combinatorial treatment synergistically induced functional transformation of PD-1+ M2-like TAMs toward a proinflammatory M1-like phenotype that gained phagocytic activity upon NF-κB and JAK/STAT inhibition, altered their migration through chemokine receptor alterations, and stimulated effector T cell proliferation. Lenalidomide was more effective than anti–PD-L1 in downregulation of the immunosuppressive IL-10, leading to decreased expression of both PD-1 and PD-L1. Overall, PD-1+ M2-like TAMs play an immunosuppressive role in CTCL. Anti–PD-L1 combined with lenalidomide provides a therapeutic strategy to enhance antitumor immunity by targeting PD-1+ M2-like TAMs in the CTCL TME.

Authors

Zhen Han, Xiwei Wu, Hanjun Qin, Yate-Ching Yuan, Daniel Schmolze, Chingyu Su, Jasmine Zain, Lilach Moyal, Emmilia Hodak, James F. Sanchez, Peter P. Lee, Mingye Feng, Steven T. Rosen, Christiane Querfeld

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Figure 6

Functional changes of M2-like TAMs after lenalidomide and anti–PD-L1 Ab treatment.

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Functional changes of M2-like TAMs after lenalidomide and anti–PD-L1 Ab ...
(A) M2-like TAMs were exposed to CCL2, CCL3, and CCL4 for 6, 24, and 48 hours; migration was determined by QCM 24-well Cell Migration assay (n = 3). (B) Transwell migration assays were performed on CTCL cell line supernatant-induced TAMs that were untreated or treated with anti–PD-L1, lenalidomide, or their combination (n = 3). (C) The migration ability of CTCL cell line supernatant-induced TAMs in the same treatment conditions as in B were quantified by QCM 24-well Cell Migration assay (n = 3). (D) The expression levels of chemokine receptors were shown for M2-like TAMs before and after treatment (n = 3). (E) CTCL cell line supernatant-induced TAMs in the same treatment conditions as in B were incubated for 2 hours with fluorescent polystyrene latex beads (n = 3). (F) CTCL cell line supernatant-induced TAMs in the same treatment conditions as above were incubated with E. coli for 45 minutes. Uptake of pHrodo Green E. coli BioParticles Conjugate was analyzed using flow cytometry (n = 3). (G) A phagocytosis assay with M2-like TAMs showing that anti–PD-L1 and lenalidomide treatment enhanced phagocytosis of cancer cells (n = 3). (H) CD4+ or CD8+ T cell proliferations were assessed when they were cocultured with CTCL cell line supernatant-induced TAMs using the MTT assay. ****P < 0.0001 by 2-tailed Student’s t test. (I) MTT assay was performed for CD4+ and CD8+ T cell proliferation upon coculture with MyLa-supernatant induced TAMs in the same treatment conditions as above. B–I, each value is the mean ± SD of 3 independent experiments. H and I (CD4+ T, n = 5; CD8+ T, n = 6). For B–G and I, significant difference was determined by 1-way ANOVA and P ≤ 0.05 was considered significant. *P < 0.05; **P < 0.01; ***P < 0.001.