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 5

L2-8R-70CAR T cells enhance the antitumor response.

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L2-8R-70CAR T cells enhance the antitumor response. (A) Diagram of human...
(A) Diagram of human L2-8R-70CAR and 8R-70CAR constructs. (B) Schematic representation of the 3-in-1 functionality of L2-8R-70CAR T cells. (C) T cells derived from a donor’s PBMCs were transduced with L2-8R-70CAR T cells (n = 3), 8R-70CAR T cells (n = 1), or untransduced T cells (UTT) (n = 1), respectively. Supernatants from T cell cultures were collected over a 27-day period for LAIR2 secretion by ELISA. (D and E) GZMB and TNF-α expression in T cells was assessed by FC following coculturing of CAR T cells (1 × 105) with CD70+U87 (1 × 105) cells (n = 4) overnight. (F and G) Human (h) and mouse (m) CAR T cells specifically recognized their respective CD70-expressing tumor origins (hTumor: CD70+U87 and CD70– pGBM1; mTumor: KR158B-CD70-Luc and KR158B-Luc). (H) Experimental design. C57BL/6 splenic T cells were transduced with 8R-70CAR or L2-8R-70CAR T cells. Mice (n = 3 groups, n = 8 mice/group) received i.c. injections of 1 × 104 CD70+ mTumor cells. Two of the groups were i.v. administered 1 × 107 CAR T cells on days 0 and 3, and the third group was untreated. (I and J) The percentage of tumor increase (day 38 a treatment) and survival were monitored. (K and L) mTumor sections (day 39 after treatment, H) were stained for collagen I and collagen IV. MFI was calculated as follows: (total intensity – background)/nuclei. (M) Comparison of intratumoral T cell infiltration after treatment. IF images of intratumoral CAR T cells were visualized, showing mouse CD8, human CXCR2, and DAPI staining in tumor sections. Scale bars: 10 μm. (N) Summary of tumor samples from 3 mice (n = 3 randomly selected sections per mouse) in each treatment group, showing the percentage of CD8+CXCR2+ cells relative to total nuclei. All experiments were repeated twice. Data are represented as the mean ± SEM (C) and box-and-whiskers plots (D–G, I, and K–M). Statistical significance was determined using GEE (C, K, L, and N), the Mann-Whitney U test (D–G, and I), and the log-rank test (J). FDR correction was applied for all comparisons. **P < 0.01 and ***P < 0.001. Schematics in A, B, and H were created using BioRender.com.