Zhou Xinsheng, Liu Yixin, Liu Xuan, Song Xu, Li Sijie, Chen Peng, Jiang Xiaotao, Li Yongyin
Department of Infectious Diseases, Nanfang Hospital, Southern Medical University; State Key Laboratory of Organ Failure Research; Guangdong Provincial Key Laboratory for Prevention and Control of Major Liver Diseases, Key Laboratory of Infectious Diseases Research in South China (Southern Medical University), Ministry of Education, Guangzhou, China.
Department of Immunology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
Invest New Drugs. 2025 May 1. doi: 10.1007/s10637-025-01530-x.
Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related mortality worldwide, with poor prognosis and limited treatment options, particularly in advanced stages. Glypican-3 (GPC3) has emerged as a promising therapeutic target, but existing antibodies primarily bind its C-terminal region, where glycosylation can mask epitopes and compromise efficacy. To address this limitation, we focused on the GPC3 N-terminal region, which offers better accessibility and potential for tumor signaling regulation. We developed Pro-12, a high-affinity humanized IgG1 antibody targeting the 25-45 peptide of the GPC3 N-terminus, avoiding glycosylation interference while modulating tumor pathways. Building on Pro-12, we engineered a GPC3/CD3 bispecific antibody (BsAb) using CrossMab and Knob-into-Hole technologies. This BsAb demonstrated dual anti-tumor effects by activating immune cells and inhibiting both the Wnt/β-catenin and PI3K/AKT pathways, achieving outcomes typically requiring tri-specific antibodies. Our findings highlight the GPC3 N-terminal region as a novel therapeutic target and introduce a promising bispecific antibody approach for the treatment of GPC3-positive HCC.
肝细胞癌（HCC）仍然是全球癌症相关死亡的主要原因，预后较差且治疗选择有限，尤其是在晚期。磷脂酰肌醇蛋白聚糖-3（GPC3）已成为一个有前景的治疗靶点，但现有的抗体主要结合其C端区域，该区域的糖基化会掩盖表位并影响疗效。为了解决这一局限性，我们聚焦于GPC3的N端区域，该区域具有更好的可及性以及调节肿瘤信号传导的潜力。我们开发了Pro-12，这是一种高亲和力的人源化IgG1抗体，靶向GPC3 N端的25-45肽段，避免了糖基化干扰，同时调节肿瘤通路。基于Pro-12，我们利用CrossMab和Knob-into-Hole技术构建了一种GPC3/CD3双特异性抗体（BsAb）。这种BsAb通过激活免疫细胞以及抑制Wnt/β-连环蛋白和PI3K/AKT通路展现出双重抗肿瘤作用，实现了通常需要三特异性抗体才能达到的效果。我们的研究结果突出了GPC3 N端区域作为一个新的治疗靶点，并引入了一种有前景的双特异性抗体方法用于治疗GPC3阳性的HCC。
