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
View: Text | PDF
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

×

Figure 4

CTCL cell line supernatant induces PD-1+ M2-like TAMs in vitro.

Options: View larger image (or click on image) Download as PowerPoint
CTCL cell line supernatant induces PD-1+ M2-like TAMs in vitro. Healthy ...
Healthy CD14+ monocytes were cultured for 72 hours in CTCL cell line–conditioned medium. (A) The expression of CD80, CD163, CD206, PD-1, and PD-L1 was detected in TAMs induced by CTCL cell line supernatant using flow cytometry. The histograms were representative of 3 independent experiments. The lighter shades at the top represent the fluorescence intensity of the untreated controls, and the darker shades at the bottom represent the fluorescence intensity of the isotype controls. (B) IL-1β, CXCL-10, CXCL-11, and IL-10 mRNA levels were assessed in total CD14+ cells cultured with or without CTCL cell line supernatant. Data are presented as mean ± SD from 3 biological replicates. **P < 0.01, ***P < 0.001, ****P < 0.0001, by 2-tailed Student’s t test. (C) Protein expression of p-IKKα/β, IKKα, IKKβ, p–NF-κB, IKBα, p-IKBα, and NF-κB and (D) JAK3, STATs, and SOCSs in lysates from TAMs induced by CM compared with expression from monocytes without CM assessed by Western blotting using GAPDH as a loading control (Ctrl, control; CM, CTCL cell line supernatant). One representative image of 3 independent samples per group is shown. (E) The hallmark pathway analysis of differentially expressed genes in CTCL (n = 45) as compared with healthy controls (n = 45). Data are representative of 3 independent experiments. For each pathway shown, the difference between groups had an FDR q value less than 0.05. (F) RNA-Seq gene-level analysis plots reveal the expression levels of JAK3 and NF-κB genes in CTCL (n = 45) and healthy controls (n = 3). The green dots indicate plaque, yellow dots indicate patch, purple dots represent tumor, blue dots show SS, and black dots show normal. Data are representative of 3 independent experiments. ****P < 0.0001, by 2-tailed Student’s t test.