Epithelial cell -derived microvesicles: A safe delivery platform of CRISPR/Cas9 conferring synergistic anti-tumor effect with sorafenib

• Published in: Experimental cell research 2020-07, Vol.392 (2), p.112040-112040, Article 112040
• Main Authors: He, Cong, Jaffar Ali, Doulathunnisa, Xu, Huantian, Kumaravel, Subhashree, Si, Ke, Li, Yumin, Sun, Bo, Ma, Junqing, Xiao, Zhongdang
• Format: Article
• Language: English
• Subjects: Animals; Antineoplastic Agents - pharmacology; Apoptosis; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - pathology; Carcinoma, Hepatocellular - therapy; Cell Proliferation; Cell-Derived Microparticles - metabolism; Cell-Derived Microparticles - pathology; Combination therapy; Combined Modality Therapy; CRISPR-Cas Systems; CRISPR/Cas9; Epithelial Cells - metabolism; Epithelial Cells - pathology; Female; Gene Editing; HEK293; HepG2; Humans; Liver Neoplasms - genetics; Liver Neoplasms - pathology; Liver Neoplasms - therapy; Mice; Mice, Inbred BALB C; Mice, Nude; Microvesicles; ras GTPase-Activating Proteins - antagonists & inhibitors; ras GTPase-Activating Proteins - genetics; Sorafenib; Sorafenib - pharmacology; Tumor Cells, Cultured; Xenograft Model Antitumor Assays
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2020.112040

Safe and efficient intracellular delivery of CRISPR/Cas9 is a key step for effective therapeutic genome editing in a wide range of diseases. This remains challenging due to multiple drawbacks of the currently available vehicles. Here we report that epithelial cell -derived microvesicles (MVs) function as safe and natural carriers for efficient delivery of CRISPR/Cas9 to treat cancer. In our study, compared to epithelial cell -derived MVs, cancer -derived MVs were quickly absorbed intracellularly by recipient cancer cells in vitro and showed selective accumulation in tumors of HepG2 xenografts in vivo, due to their cancer cell tropism dependent targeting. Surprisingly, synergistic anti-tumor effect of sgIQ 1.1 loaded Cas9MVs/HEK293 + sorafenib was better than sgIQ 1.1 + Cas9MVs/HepG2 + sorafenib in vitro. In addition, qPCR results showed that miR-21 and miR-181a expression were upregulated in HepG2 cells treated with cancer cell -derived MVs that might support the cancer progression. Further, treatment of HepG2 xenografts with sgIQ 1.1 loaded Cas9MVs/HEK293 showed enhanced anti-cancer effect than sgIQ 1.1 + Cas9MVs/HepG2. Therefore, we conclude that normal cells -derived MVs can act as better and safe natural delivery systems for cancer therapeutics in the future. Scheme of a combination therapy in liver cancer cells. Epithelial cell -derived Cas9MVs loaded with sgIQ 1.1 targeting IQGAP1 enhanced inhibition of cancer cell propagation when combined with sorafenib. sgIQ 1.1 was loaded into Cas9MVs/HepG2 and Cas9MVs/HEK293 via electroporation. Tumor -derived MVs (Cas9MVs/HepG2) accelerate their accumulation within liver cancer HepG2 cells, compared to epithelial cell -derived MVs (Cas9MVs/HEK293) because of their cancer cell tropism. Still, irrespective of their cell tropism, epithelial cell -derived Cas9MVs loaded with sgIQ 1.1 give more synergistic apoptosis effect, may be due to the possibility of tumor-derived MVs to act as carriers for cancer related molecules. [Display omitted]