Synergistic effects of carboxymethyl-hexanoyl chitosan, cationic polyurethane-short branch PEI in miR122 gene delivery: accelerated differentiation of iPSCs into mature hepatocyte-like cells and improved stem cell therapy in a hepatic failure model

Yueh Chien; Yuh-Lih Chang; Hsin-Yang Li; Mikael Larsson; Wai-Wah Wu; Chian-Shiu Chien; Chien-Ying Wang; Pen-Yuan Chu; Kuan-Hsuan Chen; Wen-Liang Lo; Shih-Hwa Chiou; Yuan-Tzu Lan; Teh-Ia Huo; Shou-Dong Lee; Pin-I Huang

doi:10.1016/j.actbio.2014.11.018

Synergistic effects of carboxymethyl-hexanoyl chitosan, cationic polyurethane-short branch PEI in miR122 gene delivery: accelerated differentiation of iPSCs into mature hepatocyte-like cells and improved stem cell therapy in a hepatic failure model

Acta Biomater. 2015 Feb:13:228-44. doi: 10.1016/j.actbio.2014.11.018. Epub 2014 Nov 21.

Authors

Yueh Chien¹, Yuh-Lih Chang¹, Hsin-Yang Li², Mikael Larsson³, Wai-Wah Wu⁴, Chian-Shiu Chien¹, Chien-Ying Wang⁵, Pen-Yuan Chu⁶, Kuan-Hsuan Chen⁷, Wen-Liang Lo⁸, Shih-Hwa Chiou⁹, Yuan-Tzu Lan¹⁰, Teh-Ia Huo¹¹, Shou-Dong Lee¹², Pin-I Huang¹³

Affiliations

¹ Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan; Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan.
² Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; Department of Obstetrics and Gynecology, Taipei Veterans General Hospital, Taipei, Taiwan.
³ Ian Wark Research Institute, University of South Australia, Mawson Lakes Campus, Mawson Lakes, SA 5095, Australia.
⁴ Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; Division of Gastroenterology, Department of Medicine, Cheng Hsin General Hospital, Taipei, Taiwan.
⁵ Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan.
⁶ Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; Laryngology and Head and Neck Surgery, Department of Otolaryngology, Taipei Veterans General Hospital, Taipei, Taiwan.
⁷ Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; Department of Pharmacy, Taipei Veterans General Hospital, Taipei, Taiwan.
⁸ Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; Division of Oral and Maxillofacial Surgery, Department of Stomatology, Taipei Veterans General Hospital, Taipei, Taiwan.
⁹ Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan; Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan.
¹⁰ Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan. Electronic address: ytlan@vghtpe.gov.tw.
¹¹ Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan; Division of Gastroenterology, Department of Internal Medicine, Taipei Veterans General Hospital, Taipei, Taiwan.
¹² Division of Gastroenterology, Department of Medicine, Cheng Hsin General Hospital, Taipei, Taiwan.
¹³ Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; Division of Radiation Oncology, Department of Oncology, Taipei Veterans General Hospital, Taipei, Taiwan. Electronic address: pihuang@vghtpe.gov.tw.

PMID: 25463491
DOI: 10.1016/j.actbio.2014.11.018

Abstract

MicroRNA122 (miR122), a liver-specific microRNA, plays critical roles in homeostatic regulation and hepatic-specific differentiation. Induced pluripotent stem cells (iPSCs) have promising potential in regenerative medicine, but it remains unknown whether non-viral vector-mediated miR122 delivery can enhance the differentiation of iPSCs into hepatocyte-like cells (iPSC-Heps) and rescue thioacetamide-induced acute hepatic failure (AHF) in vivo. In this study, we demonstrated that embedment of miR122 complexed with polyurethane-graft-short-branch polyethylenimine copolymer (PU-PEI) in nanostructured amphiphatic carboxymethyl-hexanoyl chitosan (CHC) led to dramatically enhanced miR122 delivery into human dental pulp-derived iPSCs (DP-iPSCs) and facilitated these DP-iPSCs to differentiate into iPSC-Heps (miR122-iPSC-Heps) with mature hepatocyte functions. Microarray and bioinformatics analysis further indicated that CHC/PU-PEI-miR122 promoted the gene-signature pattern of DP-iPSCs to shift into a liver-specific pattern. Furthermore, intrahepatic delivery of miR122-iPSC-Heps, but not miR-Scr-iPSC-Heps, improved liver functions and rescued recipient survival, and CHC-mediated delivery showed a better efficacy than that using phosphate buffered saline as a delivery vehicle. In addition, these transplanted miR122-iPSC-Heps remained viable and could produce circulatory albumin for 4 months. Taken together, our findings demonstrate that non-viral delivery of miR122 shortens the time of iPSC differentiation into hepatocytes and the delivery of miR122-iPSC-Heps using CHC as a vehicle exhibited promising hepatoprotective efficacy in vivo. miR122-iPSC-Heps may represent a feasible cell source and provide an efficient and alternative strategy for hepatic regeneration in AHF.

Keywords: Acute hepatic failure; Induced pluripotent stem cells; Nano structured chitosan; miR122.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation* / drug effects
Cell Differentiation* / genetics
Chitosan / analogs & derivatives*
Chitosan / pharmacology
Disease Models, Animal
Gene Transfer Techniques*
Hepatocytes* / metabolism
Hepatocytes* / transplantation
Heterografts
Humans
Induced Pluripotent Stem Cells / metabolism*
Liver Failure / genetics
Liver Failure / metabolism
Liver Failure / pathology
Liver Failure / therapy*
Male
Mice
Mice, Inbred BALB C
Mice, Nude
MicroRNAs* / biosynthesis
MicroRNAs* / genetics
Polyurethanes / pharmacology*

Substances

MIRN122 microRNA, human
MicroRNAs
Polyurethanes
carboxymethyl-hexanoyl chitosan
Chitosan