Biomedical Polymers Laboratory and Jiangsu Key Laboratory of Advanced Functional Polymer Design and Application, College of Chemistry, Chemical Engineering and Materials Science, Soochow University , Suzhou 215123, People's Republic of China.
Department of Neurosurgery, The Second Affiliated Hospital of Soochow University , Suzhou 215004, People's Republic of China.
ACS Appl Mater Interfaces. 2017 Jul 19;9(28):24140-24147. doi: 10.1021/acsami.7b06879. Epub 2017 Jul 10.
Protein drugs with intracellular targets like Granzyme B (GrB) have demonstrated great proliferative inhibition activity in cancer cells. Their clinical translation, however, relies on the development of safe, efficient, and selective protein-delivery vehicles. Here, we report that epidermal growth factor receptor (EGFR) and CD44 dual-targeted multifunctional hyaluronic acid nanogels (EGFR/CD44-NGs) boost protein delivery to ovarian and breast cancers in vitro and in vivo. EGFR/CD44-NGs obtained via nanoprecipitation and photoclick chemistry from hyaluronic acid derivatives with tetrazole, GE11 peptide/tetrazole, and cystamine methacrylate groups had nearly quantitative loading of therapeutic proteins like cytochrome C (CC) and GrB, a small size of ca. 165 nm, excellent stability in serum, and fast protein release under a reductive condition. Flow cytometry assays showed that EGFR/CD44-NGs exhibited over 6-fold better uptake in CD44 and EGFR-positive SKOV-3 ovarian cancer cells than CD44-NGs. In accordance, GrB-loaded EGFR/CD44-NGs (GrB-EGFR/CD44-NGs) displayed enhanced caspase activity and growth inhibition in SKOV-3 cells as compared to GrB-loaded CD44-NGs (GrB-CD44-NGs) control. Intriguingly, the therapeutic studies in SKOV-3 human ovarian carcinoma and MDA-MB-231 human breast tumor xenografted in nude mice revealed that GrB-EGFR/CD44-NGs at a low dose of 3.85 nmol GrB equiv/kg induced nearly complete growth suppression of both tumors, which was obviously more effective than GrB-CD44-NGs, without causing any adverse effects. EGFR and CD44 dual-targeted multifunctional hyaluronic acid nanogels have appeared as a safe and efficacious platform for cancer protein therapy.
具有细胞内靶点的蛋白质药物，如 Granzyme B（GrB），在癌细胞中显示出很强的增殖抑制活性。然而，它们的临床转化依赖于安全、高效和选择性的蛋白质递送载体的发展。在这里，我们报告表皮生长因子受体（EGFR）和 CD44 双重靶向多功能透明质酸纳米凝胶（EGFR/CD44-NGs）能够增强蛋白质在体外和体内对卵巢癌和乳腺癌的递送。通过纳米沉淀和光点击化学从具有叠氮基团的透明质酸衍生物、GE11 肽/叠氮基团和半胱氨酸甲基丙烯酰胺基团制备的 EGFR/CD44-NGs，能够近乎定量地负载细胞色素 C（CC）和 GrB 等治疗性蛋白质，粒径约为 165nm，在血清中具有极好的稳定性，并在还原条件下快速释放蛋白质。流式细胞术分析表明，与 CD44-NGs 相比，EGFR/CD44-NGs 在 CD44 和 EGFR 阳性 SKOV-3 卵巢癌细胞中的摄取量增加了 6 倍以上。相应地，与负载 GrB 的 CD44-NGs（GrB-CD44-NGs）对照组相比，负载 GrB 的 EGFR/CD44-NGs（GrB-EGFR/CD44-NGs）在 SKOV-3 细胞中显示出增强的半胱天冬酶活性和生长抑制。有趣的是，在裸鼠移植的 SKOV-3 人卵巢癌和 MDA-MB-231 人乳腺癌肿瘤的治疗研究中，发现低剂量 3.85nmol GrB 当量/公斤的 GrB-EGFR/CD44-NGs 几乎完全抑制了两种肿瘤的生长，其效果明显优于 GrB-CD44-NGs，而没有引起任何不良反应。EGFR 和 CD44 双重靶向多功能透明质酸纳米凝胶已成为癌症蛋白质治疗的一种安全有效的平台。
