Pristimerin enhances recombinant adeno-associated virus vector-mediated transgene expression in human cell lines in vitro and murine hepatocytes in vivo |
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| Institution: | 1. Changhai Hospital of Traditional Chinese Medicine, Second Military Medical University, Shanghai 200433, China;2. Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, Florida 32611, USA;3. Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, Florida 32611, USA;4. Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;5. Department of Orthopaedics and Rehabilitation, University of Florida College of Medicine, Gainesville, Florida 32611, USA;6. Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, Florida 32611, USA;7. Genetics Institute, University of Florida College of Medicine, Gainesville, Florida 32611, USA;8. Shands Cancer Center, University of Florida College of Medicine, Gainesville, Florida 32611, USA;1. School of Pharmacy, Lanzhou University, Lanzhou 730000, China;2. Gansu Academy of Agricultural Sciences, Lanzhou 730000, China;3. Gansu Institute for Drug Control, Lanzhou 730000, China;4. Key Laboratory of Biochemistry and Molecular Biology in Universities of Shandong Province, Weifang University, Weifang 261061, China;1. Changhai Hospital of Traditional Chinese Medicine, Second Military Medical University, Shanghai 200433, China;2. Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;3. Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, Florida 32611, USA;4. Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, Florida 32611, USA;5. Genetics Institute, University of Florida College of Medicine, Gainesville, Florida 32611, USA;6. Shands Cancer Center, University of Florida College of Medicine, Gainesville, Florida 32611, USA;7. Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, Florida 32611, USA;1. State Key Laboratory of Genetic Engineering and Engineering Research Center of Gene Technology (Ministry of Education), School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200438, China;2. Department of Clinical Laboratory, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang Province, China;3. Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province, Wenzhou 325000, Zhejiang Province, China;4. Yantai Fuheng Biological Technology Co., Ltd., Yantai 264006, Shandong Province, China;1. Department of Urology, University Hospital Charite, 10117 Berlin, Germany;2. Drug Metabolism and Toxicology Section, Department of Biochemistry, University of Ibadan, Ibadan 20005, Nigeria;3. Membrane Biochemistry and Biophysics Section, Department of Biochemistry, University of Ibadan, Ibadan 20005, Nigeria;4. Institute of Physiology, University Hospital Charite, 10117 Berlin, Germany;5. Berlin Institute for Urologic Research, 10117 Berlin, Germany |
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| Abstract: | OBJECTIVE:In the present study,we systemically evaluated the ability of two bioactive compounds from traditional Chinese medicine,celastrol and pristimerin,to enhance recombinant adeno-associated virus(rAAV) serotype vector-mediated transgene expression both in human cell lines in vitro,and in murine hepatocytes in vivo.METHODS:Human cell lines were infected with rAAV vectors with either mock treatment or treatment with celastrol or pristimerin.The transgene expression,percentage of nuclear translocated viral genomes and the ubiquitination of intracellular proteins were investigated post-treatment.In addition,nonobese diabetic/severe combined immunodeficient gamma(NSG) mice were tail vain-injected with rAAV vectors and co-administered with either dimethyl sulfoxide,celastrol,pristimerin or a positive control,bortezomib.The transgene expression in liver was detected and compared over time.RESULTS:We observed that treatment with pristimerin,at as low as 1 umol/L concentration,significantly enhanced rAAV2 vector-mediated transgene expression in vitro,and intraperitoneal coadministration with pristimerin at 4 mg/(kgd) for 3 d dramatically facilitated viral transduction in murine hepatocytes in vivo.The transduction efficiency of the tyrosine-mutant rAAV2 vectors as well as that of rAAV8 vectors carrying oversized transgene cassette was also augmented significantly by pristimerin.The underlying molecular mechanisms by which pristimerin mediated the observed increase in the transduction efficiency of rAAV vectors include both inhibition of proteasomal degradation of the intracellular proteins and enhanced nuclear translocation of the vector genomes.CONCLUSION:These studies suggest the potential beneficial use of pristimerin and pristimerincontaining herb extract in future liver-targeted gene therapy with rAAV vectors. |
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| Keywords: | celastrol pristimerin adeno-associated viral vector proteosomal inhibitor gene therapy |
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