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AXL antibody and AXL-ADC mediate antitumor efficacy via targeting AXL in tumor-intrinsic epithelial-mesenchymal transition and tumor-associated M2-like macrophage

Acta Pharmacologica Sinica (2023)Cite this article

Abstract

The receptor tyrosine kinase AXL is an emerging driver of cancer recurrence, while its molecular mechanism remains unclear. In this study we investigated how AXL regulated the disease progression and poor prognosis in non-small cell lung cancer (NSCLC) and triple negative breast cancer (TNBC). We performed AXL transcriptome analysis from TCGA datasets, and found that AXL expression was significantly elevated in NSCLC and TNBC correlating with poor prognosis, epithelial-mesenchymal transition (EMT) and immune-tolerant tumor microenvironment (TME). Knockdown of AXL or treatment with two independent AXL antibodies (named anti-AXL and AXL02) all diminished cell migration and EMT in AXL-high expressing NSCLC and TNBC cell lines. In a mouse model of 4T1 TNBC, administration of anti-AXL antibody substantially inhibited lung metastases formation and growth, accompanied by reduced downstream signaling activation, EMT and proliferation index, as well as an increased apoptosis and activated anti-tumor immunity. We found that AXL was abundantly activated in tumor nodule-infiltrated M2-macrophages. A specific anti-AXL antibody blocked bone marrow-derived macrophage (BMDM) M2-polarization in vitro. Targeting of AXL in M2-macrophage in addition to tumor cell substantially suppressed CSF-1 production and eliminated M2-macrophage in TME, leading to a coordinated enhancement in both the innate and adaptive immunity reflecting M1-like macrophages, mature dendritic cells, cytotoxic T cells and B cells. We generated a novel and humanized AXL-ADC (AXL02-MMAE) employing a site-specific conjugation platform. AXL02-MMAE exerted potent cytotoxicity against a panel of AXL-high expressing tumor cell lines (IC50 < 0.1 nmol/L) and suppressed in vivo growth of multiple NSCLC and glioma tumors (a minimum efficacy dose<1 mg/kg). Compared to chemotherapy, AXL02-MMAE achieved a superior efficacy in regressing large sized tumors, eliminated AXL-H tumor cell-dependent M2-macrophage infiltration with a robust accumulation of inflammatory macrophages and mature dendritic cells. Our results support AXL-targeted therapy for treatment of advanced NSCLC and TNBC.

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Fig. 1: AXL is elevated in NSCLC and TNBC and correlates with poor survival, EMT and immunosuppressive tumor environment.
Fig. 2: Characterization of AXL-high tumor models and response to anti-AXL treatment in vitro.
Fig. 3: AXL inhibition reduces lung metastasis and induces antitumor immunity in a syngeneic mouse model of TNBC.
Fig. 4: AXL-targeted inhibition of M2 macrophage polarization and function in tumor environment.
Fig. 5: AXL inhibition impairs the M2 polarization of macrophages in vitro.
Fig. 6: Discovery of highly potent and humanized anti-AXL antibody and AXL-ADC.
Fig. 7: Potent in vivo antitumor efficacy of novel AXL-ADC in athymic mice.
Fig. 8: AXL-ADC induces a dramatic tumor regression and suppression of tumor associated M2-like macrophages in athymic mice.

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Acknowledgements

This work was supported by Fudan University (EZF301002), The National Natural Science Foundation of China (81373442), NST Major Project of China (2018ZX09711002-008) and NBR 973 Program of China (2013CB932500). The authors thank Animal Facility, Instrument Center, School of Pharmacy, Fudan University and the CDSER/SIMM facility for study support.

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Authors and Affiliations

  1. Department of Pharmacology, Fudan University School of Pharmacy, Shanghai, 201203, China

    Jin-peng Pei, Yue Wang, Liang Liu, Yu Zhang, Rui Jin, Zhi-qiang Ren, Yan Deng & Ker Yu

  2. Department of Medicinal Chemistry, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China

    Lan-ping Ma, Xin Wang, Jing-kang Shen & Tao Meng

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Contributions

TM, JKS, KY designed research. JPP, YW, TM, LPM, XW, LL, YZ, ZQR, YD performed research. JPP, RJ, LPM, TM contributed new reagents or analytic tools. JPP, YW, TM analyzed data. JPP, YW, TM, KY wrote the paper.

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Correspondence to Jing-kang Shen, Tao Meng or Ker Yu.

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JPP, LPM, XW, RJ, JKS, TM and KY are listed as co-inventor in WO/2019/218944. The other authors declare no conflict of interest.

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Pei, Jp., Wang, Y., Ma, Lp. et al. AXL antibody and AXL-ADC mediate antitumor efficacy via targeting AXL in tumor-intrinsic epithelial-mesenchymal transition and tumor-associated M2-like macrophage. Acta Pharmacol Sin (2023). https://doi.org/10.1038/s41401-022-01047-6

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Keywords

  • AXL
  • NSCLC
  • TNBC
  • M2-macrophage
  • antibody-drug conjugate

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