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Allosteric inhibition of lysyl oxidase–like-2 impedes the development of a pathologic microenvironment

Abstract

We have identified a new role for the matrix enzyme lysyl oxidase–like-2 (LOXL2) in the creation and maintenance of the pathologic microenvironment of cancer and fibrotic disease. Our analysis of biopsies from human tumors and fibrotic lung and liver tissues revealed an increase in LOXL2 in disease-associated stroma and limited expression in healthy tissues. Targeting LOXL2 with an inhibitory monoclonal antibody (AB0023) was efficacious in both primary and metastatic xenograft models of cancer, as well as in liver and lung fibrosis models. Inhibition of LOXL2 resulted in a marked reduction in activated fibroblasts, desmoplasia and endothelial cells, decreased production of growth factors and cytokines and decreased transforming growth factor-β (TGF-β) pathway signaling. AB0023 outperformed the small-molecule lysyl oxidase inhibitor β-aminoproprionitrile. The efficacy and safety of LOXL2-specific AB0023 represents a new therapeutic approach with broad applicability in oncologic and fibrotic diseases.

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Figure 1: LOXL2 localization in the stroma of diverse tumor types.
Figure 2: In vitro characterization of inhibitory monoclonal antibody AB0023.
Figure 3: Effects of AB0023 on fibroblast activation and growth factor abundance in the tumor microenvironment.
Figure 4: Comparison of AB0023 and β-APN and effects of AB0023 on metastatic burden in xenograft models.
Figure 5: Effects of AB0023 in a liver fibrosis model.
Figure 6: Effects of AB0023 in bleomycin-induced lung fibrosis.

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Acknowledgements

We thank S. Lyle for guidance and interpretation of tissue pathology and J. Adamkewicz, S. Lyle, M. Longaker and F. McCormick for their review of this manuscript. We thank G. Rosen and J. Belperio for guidance with bleomycin-induced lung fibrosis studies and G. Gurtner for help with wound-healing study design. We thank J. Tambaoan for program support.

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V.B.-H. and R.S. performed immunohistochemistry and histology on human and mouse tissues and all associated analyses, with V.B.-H. leading the immunohistochemistry group, and they both participated in manuscript preparation. V.B.-H. performed EMT studies, D.M. performed transcript analyses, managed the tissue collection, designed the wound-healing model and participated in manuscript preparation, S.M. performed cloning and expression and participated in EMT experiments and manuscript preparation, H.M.R. performed data analysis, IC50 studies and antibody characterization, M.O. performed immunohistochemistry and analysis for the liver fibrosis study, A.M. and M.V. performed tension experiments and antibody characterization, A.M. participated in manuscript preparation, H.G. performed immunohistochemistry analysis for the SKOV3 study and participated in manuscript preparation, C.W. performed transcript analysis, C.A.G., A.C.V., B.J., D.B. and D.T. generated, characterized and quality controlled all antibodies and proteins under the leadership of C.A.G., J.G. performed tissue ELISA, S.Z.-E. contributed to antibody characterization and performed the microvessel density analysis, A.H. supervised the MDA-MB-435 mouse studies, S.O. assisted with the management of contract research groups, D.T. participated in toxicology studies and manuscript preparation, G.N. developed the Y698F mutant and contributed to oncology studies and manuscript preparation, P.V.V. participated in experimental design and manuscript preparation and V.S. designed the metastasis, fibrosis and toxicology animal studies and analyses, supervised the experimental work and wrote the paper.

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Correspondence to Victoria Smith.

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As current or former employees of Arresto BioSciences, V.B.-H., R.S., D.M., S.M., H.M.R., M.O., A.M., M.V., H.G., C.W., C.A.G., A.C.V., B.J., D.B., D.T., A.H., S.O., D.T., P.V.V. & V.S. have an equity stake in the company.

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Barry-Hamilton, V., Spangler, R., Marshall, D. et al. Allosteric inhibition of lysyl oxidase–like-2 impedes the development of a pathologic microenvironment. Nat Med 16, 1009–1017 (2010). https://doi.org/10.1038/nm.2208

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