Wang Minghai, Ma Qi, Suthe Sreedhar Reddy, Hudson Rachel E, Pan Jing-Ying, Mikelis Constantinos, Zhu Miao-Jin, Wu Zhi-Gang, Shi Dan-Rong, Yao Hang-Ping
State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious diseases, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003, China.
Department of Pharmaceutical Sciences, Jerry H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, Amarillo, 79106, TX, USA.
Acta Pharmacol Sin. 2025 May;46(5):1375-1389. doi: 10.1038/s41401-024-01458-7. Epub 2025 Jan 21.
Cancer heterogeneity, characterized by diverse populations of tumorigenic cells, involves the occurrence of differential phenotypes with variable expressions of receptor tyrosine kinases. Aberrant expressions of mesenchymal-epithelial transition (MET) and recepteur d'origine nantais (RON) receptors contribute to the phenotypic heterogeneity of cancer cells, which poses a major therapeutic challenge. This study aims to develop a dual-targeting antibody-drug conjugate (ADC) that can act against both MET and RON for treating cancers with high phenotypic heterogeneity. Through immunohistochemical staining, we show that MET and RON expressions are highly heterogeneous with differential combinations in more than 40% of pancreatic and triple-negative breast cancer cases. This expressional heterogeneity provides the rationale to target both receptors for cancer therapy. A humanized bispecific monoclonal antibody specific to both MET and RON (PCMbs-MR) is generated through IgG recombination using monoclonal antibody sequences specific to MET and RON, respectively. Monomethyl auristatin E is conjugated to PCMbs-MR to generate a dual-targeting ADC (PCMdt-MMAE), with a drug-to-antibody ratio of 4:1. Various cancer cell lines were used to determine PCMdt-MMAE-mediated biological activities. The efficacy of PCMdt-MMAE in vivo is evaluated using multiple xenograft tumor models. PCMdt-MMAE shows a favorable pharmacokinetic profile, with a maximum tolerated dose of ~30 mg/kg in mice. Toxicological studies using Sprague-Dawley rats reveal that PCMdt-MMAE is relatively safe with slight-to-moderate, temporary, and reversible adverse events. Functionally, PCMdt-MMAE induces a robust internalization of both MET and RON and causes a large-scale cell death in cancer cell lines exhibiting MET and RON heterogeneous co-expressions. Both in vitro and in vivo studies demonstrate that the dual-targeting approach in the form of an ADC is highly effective with a long-lasting effect against tumors exhibiting MET/RON heterogeneous phenotypes. Hence, we can suggest that a dual-targeting ADC specific to both MET and RON can be employed as a novel therapeutic strategy for tumors with expressional phenotypic heterogeneity.
癌症异质性以多种致瘤细胞群为特征，涉及具有不同受体酪氨酸激酶表达的差异表型的出现。间充质-上皮转化（MET）和南特起源受体（RON）受体的异常表达导致癌细胞的表型异质性，这构成了重大的治疗挑战。本研究旨在开发一种双靶向抗体-药物偶联物（ADC），其可同时作用于MET和RON，用于治疗具有高表型异质性的癌症。通过免疫组织化学染色，我们发现MET和RON表达在超过40%的胰腺癌和三阴性乳腺癌病例中高度异质且组合不同。这种表达异质性为针对两种受体进行癌症治疗提供了理论依据。通过分别使用针对MET和RON的单克隆抗体序列进行IgG重组，产生了一种对MET和RON均特异的人源化双特异性单克隆抗体（PCMbs-MR）。将单甲基澳瑞他汀E与PCMbs-MR偶联，生成双靶向ADC（PCMdt-MMAE），药物与抗体比例为4:1。使用多种癌细胞系来确定PCMdt-MMAE介导的生物学活性。使用多种异种移植肿瘤模型评估PCMdt-MMAE在体内的疗效。PCMdt-MMAE显示出良好的药代动力学特征，在小鼠中的最大耐受剂量约为30mg/kg。使用斯普拉格-道利大鼠进行的毒理学研究表明，PCMdt-MMAE相对安全，具有轻微至中度、暂时且可逆的不良事件。在功能上，PCMdt-MMAE诱导MET和RON的强烈内化，并在表现出MET和RON异质共表达的癌细胞系中导致大规模细胞死亡。体外和体内研究均表明，以ADC形式的双靶向方法对表现出MET/RON异质表型的肿瘤具有高效且持久的抗肿瘤作用。因此，我们可以认为，一种对MET和RON均特异的双靶向ADC可作为具有表达表型异质性肿瘤的新型治疗策略。
