Targeting immune checkpoint LAIR1 with antibody blockade or 3-in-1 CAR T cells enhances antitumor response
Haipeng Tao, Dongjiang Chen, Changlin Yang, Duy T. Nguyen, Georges Abboud, Ruixuan Liu, Tianyi Liu, Avirup Chakraborty, Alicia Y. Hou, Nicole A. Petit, Muhammad Abbas, Robert W. Davis, Janie Zhang, Christina Von Roemeling, Mohammed O. Gbadamosi, Linchun Jin, Tongjun Gu, Tuo Lin, Pengchen Wang, Alfonso Pepe, Diego Ivan Pedro, Hector R. Mendez-Gomez, Chao Xie, Aida Karachi, Frances Weidert, Dan Jin, Chenggang Wang, Kaytora Long-James, Elizabeth K. Molchan, Paul Castillo, John A. Ligon, Ashley P. Ghiaseddin, Elias J. Sayour, Maryam Rahman, Loic P. Deleyrolle, Betty Y.S. Kim, Duane A. Mitchell, W. Gregory Sawyer, Jianping Huang
Haipeng Tao, Dongjiang Chen, Changlin Yang, Duy T. Nguyen, Georges Abboud, Ruixuan Liu, Tianyi Liu, Avirup Chakraborty, Alicia Y. Hou, Nicole A. Petit, Muhammad Abbas, Robert W. Davis, Janie Zhang, Christina Von Roemeling, Mohammed O. Gbadamosi, Linchun Jin, Tongjun Gu, Tuo Lin, Pengchen Wang, Alfonso Pepe, Diego Ivan Pedro, Hector R. Mendez-Gomez, Chao Xie, Aida Karachi, Frances Weidert, Dan Jin, Chenggang Wang, Kaytora Long-James, Elizabeth K. Molchan, Paul Castillo, John A. Ligon, Ashley P. Ghiaseddin, Elias J. Sayour, Maryam Rahman, Loic P. Deleyrolle, Betty Y.S. Kim, Duane A. Mitchell, W. Gregory Sawyer, Jianping Huang
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Research Article Oncology

Targeting immune checkpoint LAIR1 with antibody blockade or 3-in-1 CAR T cells enhances antitumor response

Abstract

Tumor-associated macrophages (TAMs) are abundant in the tumor microenvironment (TME) and dampen the immune response, negatively affecting patient survival. Therefore, targeting TAMs could address the limitations of current cancer treatments. However, drug development in this area remains limited. The leukocyte-associated Ig-like receptor 1 (LAIR1), also called CD305, is prominently expressed on the surface of TAMs. We have uncovered what we believe to be a previously unrecognized immunosuppressive LAIR1/factor XIII A/collagen IV pathway across various cancer types. Inhibition of LAIR1, either through knockout (Lair1–/–), antibody blockade (anti-Lair1 antibody), or a chimeric antigen receptor (CAR) design (3-in-1 CAR by combining tumor targeting, T cell trafficking, and remodeling of the immunosuppressive TME in 1 CAR construct) provided an enhanced antitumor response. LAIR1 inhibition enhanced peripheral and intratumoral CD8 memory T cell populations, induced a phenotypic shift of M2-like macrophages toward M1 macrophages, and normalized tumor collagen IV and structural components in the TME, facilitating effective tumor–T cell interactions and tumor suppression. Enhanced antitumor responses were observed when Lair1–/– or anti-Lair1 antibody was used alone or in combination with CAR T cells or when the 3-in-1 CAR T cells were used solely in tumor models resistant to chemotherapy–radiation–programmed cell death protein 1 (PD-1) blockade. These findings position LAIR1 inhibition as a promising strategy for cancer immunotherapies.

Authors

Haipeng Tao, Dongjiang Chen, Changlin Yang, Duy T. Nguyen, Georges Abboud, Ruixuan Liu, Tianyi Liu, Avirup Chakraborty, Alicia Y. Hou, Nicole A. Petit, Muhammad Abbas, Robert W. Davis, Janie Zhang, Christina Von Roemeling, Mohammed O. Gbadamosi, Linchun Jin, Tongjun Gu, Tuo Lin, Pengchen Wang, Alfonso Pepe, Diego Ivan Pedro, Hector R. Mendez-Gomez, Chao Xie, Aida Karachi, Frances Weidert, Dan Jin, Chenggang Wang, Kaytora Long-James, Elizabeth K. Molchan, Paul Castillo, John A. Ligon, Ashley P. Ghiaseddin, Elias J. Sayour, Maryam Rahman, Loic P. Deleyrolle, Betty Y.S. Kim, Duane A. Mitchell, W. Gregory Sawyer, Jianping Huang

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Figure 6

LAIR1 inhibition reduces tumor collagen IV by lowering FXIII-A in M2-like TAMs and MΦ.

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LAIR1 inhibition reduces tumor collagen IV by lowering FXIII-A in M2-lik...
(A and B) Collagen IV IF staining was performed on normal brain tissues from Lair1+/+ and Lair1–/– mice, and MFI was quantified. Scale bars: 10 μm. (C and D) Tumors from Lair1+/+ and Lair1–/– TB mice (KR158B-CD70-Luc) were collected 40 days after treatment (Figure 2A) and stained for collagen IV. Scale bars: 10 μm (top), 5 μm (bottom). (E and F) IF images showing collagen IV staining of tumor tissue from C57BL/6 TB mice (LLC1) treated with IgG or aLAIR1, and tumors were collected on post-treatment day 13 (Figure 4G). Scale bars: 10 μm (top), 5 μm (bottom). The 3D image reconstruction (A, C, and E) and quantification (B, D, and F) were based on 2D images (n = 3 mice/group, n = 9 sections total) were performed. (G and H) C57BL/6 TB mice (KR158B-CD70-Luc, 3/group) with or without IgG or aLAIR1 (experimental design as in Figure 4A). Tumor scRNA-Seq was performed 49 days after treatment. UMAPs illustrate immune cell clusters with population summaries (>1%) shown. (I) Heatmap of the top 5 up- and downregulated genes in all-MΦ clusters comparing the aLAIR1- versus IgG-treated groups. (J) Volcano plot highlighting DEGs in M2-like TAMs, identifying F13a1, which encodes FXIII-A. (K) BM monocytes from Lair1+/+ and Lair1–/– C57BL/6 mice were cultured with M-CSF (100 ng/mL, 7 days), followed by IL-4, IL-10, and IL-13 (20 ng/mL, 2 days) to induce M2-like MΦ. Control cells were cultured without cytokines. Cells were harvested on day 9, and FXIII-A expression was assessed by Western blotting. (L) Arg-1 MFI was measured by FC in BM-derived M2-like MΦ (n = 3~4) treated with or without 10 μg/mL murine FXIII-A for 48 hours. M2-like MΦ were gated on CD45+CD11b+F4/80+Arg-1+cells. (M and N) KR158B-CD70-Luc cells (n = 9/group) were incubated with or without 10 μg/mL FXIII-A for 24 hours, and collagen IV levels were quantified and expression imaged. Scale bars: 4 μm. All data were reproduced 2 or more times. Data are shown as box-and-whisker plots (B, D, F, H, L, and N). Statistical significance was determined by GEE (B, D, and F), a 2-sample t test (H and J), and the Mann-Whitney U test (L). FDR correction was applied (J). *P < 0.05, **P < 0.01 and ***P < 0.001.