AXL抗体和AXL-ADC通过靶向肿瘤内在的上皮-间质转化（EMT）和肿瘤相关的M2样巨噬细胞发挥抗肿瘤作用

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

影响因子:8.4

分区:医学2区 / 药学1区化学：综合2区

发表日期:2023 Jun

作者: Jin-Peng Pei, Yue Wang, Lan-Ping Ma, Xin Wang, Liang Liu, Yu Zhang, Rui Jin, Zhi-Qiang Ren, Yan Deng, Jing-Kang Shen, Tao Meng, Ker Yu

DOI: 10.1038/s41401-022-01047-6

摘要

受体酪氨酸激酶AXL是癌症复发的一个新兴驱动因子，但其分子机制尚不清楚。本研究探讨了AXL如何调控非小细胞肺癌（NSCLC）和三阴性乳腺癌（TNBC）中的疾病进展和预后差异。我们利用TCGA数据库进行AXL转录组分析，发现AXL在NSCLC和TNBC中表达显著升高，并与预后差、上皮-间质转化（EMT）及免疫耐受的肿瘤微环境（TME）相关。敲低AXL或使用两种独立的AXL抗体（命名为anti-AXL和AXL02）均能抑制AXL高表达的NSCLC和TNBC细胞系的迁移和EMT。在4T1三阴性乳腺癌小鼠模型中，给予抗-AXL抗体显著抑制肺转移的形成和生长，同时伴随下调下游信号通路激活、EMT、增殖指数的降低、细胞凋亡增加及抗肿瘤免疫的增强。我们发现AXL在肿瘤结节浸润的M2型巨噬细胞中高度激活。特异性抗-AXL抗体可在体外阻断骨髓来源巨噬细胞（BMDM）的M2极化。除了靶向肿瘤细胞外，靶向M2型巨噬细胞同样能显著抑制CSF-1的产生，消除肿瘤微环境中的M2型巨噬细胞，从而协调增强先天和获得性免疫，包括M1样巨噬细胞、成熟树突状细胞、细胞毒性T细胞和B细胞。我们开发了一种新型人源化AXL-ADC（AXL02-MMAE），采用位点特异性偶联平台。AXL02-MMAE在一系列AXL高表达的肿瘤细胞系中表现出强效细胞毒性（IC50 < 0.1 nmol/L），并抑制多种NSCLC和胶质瘤肿瘤的体内生长（最低有效剂量<1 mg/kg）。与化疗相比，AXL02-MMAE在缩小大尺寸肿瘤方面具有更优的疗效，能消除AXL高表达肿瘤细胞依赖的M2巨噬细胞浸润，并显著积累炎症性巨噬细胞和成熟树突状细胞。我们的研究支持以AXL为靶点的治疗策略，用于治疗晚期NSCLC和TNBC。

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.