Introduction: Jounce has generated cell type-specific gene signatures as a means of probing The Cancer Genome Atlas and other large datasets to identify targets that may be important immune checkpoints. Using a tumor-associated macrophage (TAM) gene signature, we have found a strong correlation and coherence between TAMs and LILRB2 (leukocyte immunoglobulin like receptor B2; ILT4) across multiple tumors types. LILRB2 is a myeloid cell surface receptor containing four extracellular immunoglobulin domains, a transmembrane domain, and three cytoplasmic immunoreceptor tyrosine-based inhibitory motifs (ITIMs). Ligation of LILRB2 on myeloid cells, via its endogenous ligands (classical MHC I molecules [e.g. HLA-A, HLA-B] and non-classical MHC I molecules [e.g. HLA-G]), provides a negative signal that inhibits stimulation of an immune response. HLA-G is recognized as an important immunosuppressive molecule playing a role in maternal-fetal tolerance and being overexpressed in cancer - often associated with advanced disease stage and poor prognosis. As tumor-associated macrophages are known to suppress the anti-cancer immune response, these findings provide rationale for targeting LILRB2.

Methods and Results: We have generated a panel of monoclonal antibodies that bind specifically to LILRB2, but not other LILR family members, and can block binding of LILRB2 to MHC I molecules (i.e. HLA-A and HLA-G). In vitro differentiated monocyte-derived macrophages (MDMs) cultured for 24h in the presence of anti-LILRB2 antibodies and lipopolysaccharide (LPS) show polarization toward a more inflammatory phenotype - secreting higher levels of TNF-α and IL-6 with decreased amounts of IL-10 and CCL2 as compared to an isotype control antibody. NanoString mRNA analysis revealed that, in the absence of LPS or any additional stimuli, MDMs cultured with anti-LILRB2 antibodies showed gene changes consistent with inflammatory or M1-like polarization of macrophages. Anti-LILRB2 antibodies were also evaluated in human tumor histoculture and induced pharmacodynamic responses consistent with macrophage and T cell activation in a variety of tumor types. While mice do not express LILRB2 specifically, they do express a LILRB-like molecule known as Pirb. Mice that are deficient in Pirb display resistance to mouse colon 38 (MC-38) tumor growth suggesting this pathway functions as immune checkpoint in cancer.

Conclusions: Based on these preclinical data, JTX-8064, a high affinity LILRB2-specific humanized antagonist monoclonal antibody, is being developed as an immunotherapeutic to reprogram suppressive macrophages within the tumor microenvironment.

Citation Format: Heather Cohen, Yasmin Hashambhoy-Ramsay, Lauren R. Pepper, Jeffrey Y. Smith, Margaret Willer, Kevin Guay, Vikki Spaulding, Kristin O'Malley, Monica Gostissa, Abha Dhaneshwar, Edward C. Stack, Alessandro Mora, Donald R. Shaffer. Preclinical evaluation of JTX-8064, an anti-LILRB2 antagonist antibody, for reprogramming tumor-associated macrophages [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5007.