A Cell-Penetrating Peptide Improves Anti-HER2 Single-Chain Variable Fragment Internalization and Antitumor Activity against HER2-Positive Breast Cancer In Vitro and In Vivo
机构：[1]MOE International Joint Laboratory for Synthetic Biology and Medicines, School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China.[2]School of Biology and Food Science, Shangqiu Normal University, Shangqiu 476000, China.[3]Guangdong Provincial Hospital of Chinese Medicine, The Second Affiliated Hospital of Guangzhou, University of Chinese Medicine, Guangzhou 510006, China.广东省中医院[4]Nanyang Medical College, Nanyang 473001, China.[5]NMPA Key Laboratory for Quality Control of Blood Products, Guangdong Institute for Drug Control, Guangzhou 510663, China.
Cell-penetrating peptides (CPPs) are invaluable tools for delivering various substances into cells by crossing biological membranes. However, the effects of cell-penetrating peptide fusion proteins on the biological activity of antibodies remain to be fully understood. Here, we engineered a recombinant protein, LP-scFv, which combines the single-chain variable region of anti-human epidermal growth factor receptor-2 with a novel and non-oxic cell-penetrating peptide as a leader peptide. The introduction of this leader peptide led to a more than twofold increase in the internalization efficiency of the single-chain antibody, as confirmed using microscopic analysis and flow cytometry. The effects of the single-chain antibodies and LP-scFv on cell viability were evaluated using the MTT assay. Both the single-chain antibodies and LP-scFv reduced the viability of BT474 and NCI-N87 cells in a dose-dependent manner while exhibiting minimal toxicity towards MCF-7 and MCF-10A cells. Further investigation into LP-scFv's mechanism revealed that the induced leader peptide does not alter the MAPK-ERK1/2 and PI3K/AKT pathways of single-chain antibodies. An enhanced antitumor activity was also confirmed in an NCI-N87 tumor xenograft model in mice with a reduction of 45.2% in tumor growth inhibition (vs. 23.1% for scFv) with a 50 mg/kg dose after orthotopic injection administration, which was equivalent to that of trastuzumab (vs. 55.7% for trastuzumab). Overall, these results indicate that LP-scFv exhibits significant permeation activity in HER2-positive cells to enhance the intracellular dose effect on antitumor activity in vitro and in vivo. This research lays the foundation for designing novel antibody-based therapies for cancer.
基金：
National Key R&D Projects [2023YFC3040700]
the Guangdong Provincial Medical Products Administration [2023ZDZ02, 2023YDZ02], the Basic and
Applied Basic Research Foundation of Guangdong Province [2021B1515120037]; the Special project
for Research and Development in Key areas of Guangdong Province [2020B010188001]; the Natural
Science Foundation of Henan Province (No. 222300420259), and the Key Scientific Research Project in
Universities of Henan Province (No. 24A310009).
语种：
外文
PubmedID：
中科院(CAS)分区：
出版当年[2023]版：
大类|2 区化学
小类|3 区生化与分子生物学3 区化学：综合
最新[2025]版：
大类|3 区化学
小类|3 区生化与分子生物学3 区化学：综合
第一作者：
第一作者机构：[1]MOE International Joint Laboratory for Synthetic Biology and Medicines, School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China.[2]School of Biology and Food Science, Shangqiu Normal University, Shangqiu 476000, China.
共同第一作者：
通讯作者：
推荐引用方式(GB/T 7714)：
Li Junmin,Zhou Yanting,Su Zhuowei,et al.A Cell-Penetrating Peptide Improves Anti-HER2 Single-Chain Variable Fragment Internalization and Antitumor Activity against HER2-Positive Breast Cancer In Vitro and In Vivo[J].Molecules (Basel, Switzerland).2024,29(6):doi:10.3390/molecules29061247.
APA：
Li Junmin,Zhou Yanting,Su Zhuowei,Li Xue,Zhang Lei&Li Shan.(2024).A Cell-Penetrating Peptide Improves Anti-HER2 Single-Chain Variable Fragment Internalization and Antitumor Activity against HER2-Positive Breast Cancer In Vitro and In Vivo.Molecules (Basel, Switzerland),29,(6)
MLA：
Li Junmin,et al."A Cell-Penetrating Peptide Improves Anti-HER2 Single-Chain Variable Fragment Internalization and Antitumor Activity against HER2-Positive Breast Cancer In Vitro and In Vivo".Molecules (Basel, Switzerland) 29..6(2024)
