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 5

Lenalidomide and anti–PD-L1 Ab reprogram M2-like TAMs in vitro.

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Lenalidomide and anti–PD-L1 Ab reprogram M2-like TAMs in vitro. (A) CTCL...
(A) CTCL cell line supernatant-induced TAMs were treated with anti–PD-L1 (10 μg/mL), lenalidomide (10 μM), or their combination for 72 hours. The expression of CD80, CD163, CD206, PD-1, and PD-L1 was detected by flow cytometry. The histograms are representative of 3 independent experiments. The shades represent the fluorescence intensity of the untreated control. (B) IL-1β, CXCL-10, CXCL-11, and IL-10 mRNA levels were assessed in total CD14+ cells cultured with CTCL cell line supernatant and treated with control, anti–PD-L1, lenalidomide, or combined anti–PD-L1 and lenalidomide. (C) IL-1β, CXCL-10, CXCL-11, and IL-10 protein levels in supernatants were assessed by ELISA in anti–PD-L1, lenalidomide, combined anti–PD-L1 and lenalidomide treatment, or untreated control. Data are representative of 3 independent experiments with mean ± SD for B and C. Significant difference was determined by 1-way ANOVA and P ≤ 0.05 was considered significant. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. (D) Western blot was utilized to determine the levels of JAK3, STATs, p–NF-κB, NF-κB, and GAPDH in lysates from CTCL cell line supernatant-induced TAMs that were untreated or treated with anti–PD-L1, lenalidomide, or their combination. Results show a representative image from 3 independent samples per group.