Linjuan Zeng1, Jingguo Li, Jiajia Li, Qiubo Zhang, Chenchen Qian, Wei Wu, Zhong Lin, Jianzhong Liang, Yinting Chen, Kaihong Huang. 1. From the *Department of Gastroenterology, The Second Affiliated Hospital of Sun Yat-sen University, Guangzhou; †Department of Oncology, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai; ‡PCFM Lab of Ministry of Education, School of Chemistry; Chemical Engineering, Sun Yat-sen University, Guangzhou; §Henan Eye Institute, Henan Eye Hospital, Zhengzhou; ∥Department of Gastroenterology, Lihuili Hospital of Ningbo Medical Center, Ningbo; ¶Department of Cardiology, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai; and #State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Guangzhou, China.
Abstract
OBJECTIVES: The objective of this study was to establish an efficient carrier for small interfering RNA (siRNA) delivery targeting pancreatic tumor cells. METHODS: A copolymer consisting of a single-chain variable fragment targeted to human CD44 variant 6 (scFv(CD44v6)) functional group conjugated to polyethylene glycol-poly-L-lysine was synthesized and assembled into micelles encapsulating the siRNAs. Flow cytometry and Western blot assays were performed to evaluate the transfection efficiency and gene-silencing effect of the siRNAs. Afterward, (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, Transwell, soft agar colony formation, and enzyme-linked immunosorbent assays were performed to evaluate the biological functions of PANC-1 cells after Kirsten rat sarcoma viral oncogene knockdown. In vivo assays were performed using a BALB/c (nu/nu) mouse model subcutaneously injected with PANC-1 xenografts. Real-time in vivo fluorescence imaging was used to monitor the tumor homing of the nanoparticles. RESULTS: The scFv(CD44v6) enabled more efficient delivery of siRNAs and exhibited enhanced gene silencing compared with nontargeted nanoparticles. Furthermore, targeted delivery of the siRNAs induced a potent inhibitory effect on cell proliferation, colony formation, invasion, and vascular endothelial growth factor production. The animal assays revealed that single-chain variable fragment nanoparticles accumulated in the tumor tissue and enhanced the inhibition of tumor growth in vivo. CONCLUSIONS: The scFv(CD44v6)-conjugated nanocarriers provide a highly efficient and safe platform for systemic gene therapy for pancreatic cancer.
OBJECTIVES: The objective of this study was to establish an efficient carrier for small interfering RNA (siRNA) delivery targeting pancreatic tumor cells. METHODS: A copolymer consisting of a single-chain variable fragment targeted to human CD44 variant 6 (scFv(CD44v6)) functional group conjugated to polyethylene glycol-poly-L-lysine was synthesized and assembled into micelles encapsulating the siRNAs. Flow cytometry and Western blot assays were performed to evaluate the transfection efficiency and gene-silencing effect of the siRNAs. Afterward, (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, Transwell, soft agar colony formation, and enzyme-linked immunosorbent assays were performed to evaluate the biological functions of PANC-1 cells after Kirsten ratsarcoma viral oncogene knockdown. In vivo assays were performed using a BALB/c (nu/nu) mouse model subcutaneously injected with PANC-1 xenografts. Real-time in vivo fluorescence imaging was used to monitor the tumor homing of the nanoparticles. RESULTS: The scFv(CD44v6) enabled more efficient delivery of siRNAs and exhibited enhanced gene silencing compared with nontargeted nanoparticles. Furthermore, targeted delivery of the siRNAs induced a potent inhibitory effect on cell proliferation, colony formation, invasion, and vascular endothelial growth factor production. The animal assays revealed that single-chain variable fragment nanoparticles accumulated in the tumor tissue and enhanced the inhibition of tumor growth in vivo. CONCLUSIONS: The scFv(CD44v6)-conjugated nanocarriers provide a highly efficient and safe platform for systemic gene therapy for pancreatic cancer.
Authors: Matthew S Strand; Bradley A Krasnick; Hua Pan; Xiuli Zhang; Ye Bi; Candace Brooks; Christopher Wetzel; Narendra Sankpal; Timothy Fleming; S Peter Goedegebuure; David G DeNardo; William E Gillanders; William G Hawkins; Samuel A Wickline; Ryan C Fields Journal: Oncotarget Date: 2019-07-30
Authors: Luman Liu; Prakash G Kshirsagar; Shailendra K Gautam; Mansi Gulati; Emad I Wafa; John C Christiansen; Brianna M White; Surya K Mallapragada; Michael J Wannemuehler; Sushil Kumar; Joyce C Solheim; Surinder K Batra; Aliasger K Salem; Balaji Narasimhan; Maneesh Jain Journal: Theranostics Date: 2022-01-01 Impact factor: 11.600