Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland.
Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland.
Int J Biol Macromol. 2024 Jan;254(Pt 1):127657. doi: 10.1016/j.ijbiomac.2023.127657. Epub 2023 Oct 28.
Breast cancer remains a significant global health challenge, necessitating the development of effective targeted therapies. This study aimed to create bispecific targeting molecules against HER2 and FGFR1, two receptors known to play crucial roles in breast cancer progression. By combining the high-affinity Affibody Z and a modified, stable form of fibroblast growth factor 2 (FGF2), we successfully generated bispecific proteins capable of simultaneously recognizing HER2 and FGFR1. Two variants were designed: Af-sFGF2 with a shorter linker and Af-lFGF2 with a longer linker between the HER2 and FGFR1-recognizing proteins. Both proteins exhibited selective binding to HER2 and FGFR1, with Af-lFGF2 demonstrating simultaneous binding to both receptors. Af-lFGF2 exhibited superior internalization compared to FGF2 in FGFR-positive cells and significantly increased internalization compared to Af in HER2-positive cells. To enhance their therapeutic potential, highly potent cytotoxic agent MMAE was conjugated to the targeting proteins, resulting in protein-drug conjugates. The bispecific Af-lFGF2-vcMMAE conjugate demonstrated exceptional cytotoxic activity against HER2-positive, FGFR-positive, and dual-positive cancer cell lines that was significantly higher compared to monospecific conjugates. These data indicate the beneficial effect of simultaneous targeting of HER2 and FGFR1 in precise anticancer medicine and contribute valuable insights into the design and potential of bispecific targeting molecules for breast cancer treatment.
乳腺癌仍然是一个重大的全球健康挑战，需要开发有效的靶向治疗方法。本研究旨在创建针对 HER2 和 FGFR1 的双特异性靶向分子，这两种受体已知在乳腺癌进展中发挥关键作用。通过结合高亲和力的 Affibody Z 和改良的、稳定形式的成纤维细胞生长因子 2（FGF2），我们成功地生成了能够同时识别 HER2 和 FGFR1 的双特异性蛋白。设计了两种变体：具有较短接头的 Af-sFGF2 和具有 HER2 和 FGFR1 识别蛋白之间较长接头的 Af-lFGF2。两种蛋白均表现出对 HER2 和 FGFR1 的选择性结合，Af-lFGF2 表现出同时与两种受体结合。与 FGFR 阳性细胞中的 FGF2 相比，Af-lFGF2 在 FGFR 阳性细胞中的内化能力更强，与 HER2 阳性细胞中的 Af 相比，内化能力显著增强。为了增强其治疗潜力，将高效细胞毒素 MMAE 缀合到靶向蛋白上，得到蛋白药物偶联物。双特异性 Af-lFGF2-vcMMAE 缀合物对 HER2 阳性、FGFR 阳性和双重阳性癌细胞系表现出卓越的细胞毒性活性，与单特异性缀合物相比，其活性显著更高。这些数据表明，同时靶向 HER2 和 FGFR1 在精确抗癌药物中的有益效果，并为乳腺癌治疗的双特异性靶向分子的设计和潜力提供了有价值的见解。
