Knockdown of hypoxia-inducible factor-1 alpha by tumor targeted delivery of CRISPR/Cas9 system suppressed the metastasis of pancreatic cancer.

Abstract

The hypoxic tumor microenvironment of pancreatic cancer contributes to the progression and metastasis of tumor cells. Downregulation of hypoxia-inducible factor-1α (HIF-1α) with CRISPR/Cas9 is a promising approach to modulate tumor microenvironment and inhibit tumor metastasis. However, the in vivo delivery of CRISPR/Cas9 system remains a challenge. In the present manuscript, a tumor targeted lipid-based CRISPR/Cas9 delivery system was developed to suppress HIF-1α. Plasmids encoding Cas9 and HIF-1α-targeting sgRNA were successfully constructed and coencapsulated in R8-dGR peptide modified cationic liposome with PTX. R8-dGR-Lip exhibited enhanced BxPC-3 cell targeting and deep penetration into tumor spheroids. R8-dGR-Lip/PTX/pHIF-1α successfully downregulated HIF-1α and its downstream molecules VEGF and MMP-9, leading to enhanced antimetastatic effects. Besides, the blockade of HIF-1α also promoted the cytotoxicity of PTX on BxPC-3 cell lines. Compared with pegylated liposomes, R8-dGR-Lip enhanced the distribution in tumor tissues. The targeted delivery of CRISPR/Cas9-HIF-1α system and PTX significantly inhibited tumor growth. More importantly, inhibition of HIF-1α suppressed the metastasis of pancreatic cancer and prolonged survival time. Since CRISPR/Cas 9-HIF-1α hardly affected HIF-1α expression in normal hepatic cells, the designed R8-dGR-Lip/PTX/pHIF-1α did not induce severe toxicity in vivo. This strategy broadened the in vivo application of CRISPR/Cas9 system. Downregulation of HIF-1α may be a feasible approach for antimetastatic therapy.

• Authors: Man Li, Hanbing Xie, Yingke Liu, Chunyu Xia, Xingli Cun, Yang Long, Xiaoxiao Chen, Miao Deng, Rong Guo, Zhirong Zhang, Qin He
• Journal: Journal of controlled release : official journal of the Controlled Release Society (J Control Release) Vol. 304 Pg. 204-215 (06 28 2019) ISSN: 1873-4995 [Electronic] Netherlands
• PMID: 31100311 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2019 Elsevier B.V. All rights reserved.

Chemical References

• HIF1A protein, human
• Hypoxia-Inducible Factor 1, alpha Subunit
• Lipids
• Liposomes
• CRISPR-Associated Protein 9

Topics

• Animals
• CRISPR-Associated Protein 9 (genetics)
• Cell Hypoxia
• Cell Line, Tumor
• Gene Knockdown Techniques
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit (genetics)
• Lipids (chemistry)
• Liposomes
• Mice
• Mice, Inbred BALB C
• Mice, Nude
• Neoplasm Metastasis (prevention & control)
• Pancreatic Neoplasms (genetics, pathology)
• Survival Rate
• Tumor Microenvironment (genetics)
• Xenograft Model Antitumor Assays