Adeno-associated virus gene therapy vector scAAVIGF-I for transduction of equine articular chondrocytes and RNA-seq analysis

Summary Objective IGF-I is one of several anabolic factors being investigated for the treatment of osteoarthritis (OA). Due to the short biological half-life, extended administration is required for more robust cartilage healing. Here we create a self-complimentary adeno-associated virus (AAV) gene...

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Bibliographic Details
Published in:Osteoarthritis and cartilage 2016-05, Vol.24 (5), p.902-911
Main Authors: Hemphill, D.D, McIlwraith, C.W, Slayden, R.A, Samulski, R.J, Goodrich, L.R
Format: Article
Language:English
Subjects:
AAV
Animals
Cartilage, Articular - cytology
Cartilage, Articular - metabolism
Chondrocytes - metabolism
Dependovirus - genetics
Enzyme-Linked Immunosorbent Assay - methods
Equine
Gene Expression Profiling - methods
Gene Expression Regulation
Gene Ontology
Gene therapy
Genetic Therapy - methods
Genetic Vectors - genetics
Horses - metabolism
IGF-I
Insulin-Like Growth Factor I - biosynthesis
Insulin-Like Growth Factor I - genetics
Joint
NextGen sequencing
Rheumatology
Sequence Analysis, RNA - methods
Transduction, Genetic
Transgenes
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Summary:Summary Objective IGF-I is one of several anabolic factors being investigated for the treatment of osteoarthritis (OA). Due to the short biological half-life, extended administration is required for more robust cartilage healing. Here we create a self-complimentary adeno-associated virus (AAV) gene therapy vector utilizing the transgene for IGF-I. Design Various biochemical assays were performed to investigate the cellular response to scAAVIGF-I treatment vs an scAAVGFP positive transduction control and a negative for transduction control culture. RNA-sequencing analysis was also performed to establish a differential regulation profile of scAAVIGF-I transduced chondrocytes. Results Biochemical analyses indicated an average media IGF-I concentration of 608 ng/ml in the scAAVIGF-I transduced chondrocytes. This increase in IGF-I led to increased expression of collagen type II and aggrecan and increased protein concentrations of cellular collagen type II and media glycosaminoglycan vs both controls. RNA-seq revealed a global regulatory pattern consisting of 113 differentially regulated GO categories including those for chondrocyte and cartilage development and regulation of apoptosis. Conclusions This research substantiates that scAAVIGF-I gene therapy vector increased production of IGF-I to clinically relevant levels with a biological response by chondrocytes conducive to increased cartilage healing. The RNA-seq further established a set of differentially expressed genes and gene ontologies induced by the scAAVIGF-I vector while controlling for AAV infection. This dataset provides a static representation of the cellular transcriptome that, while only consisting of one time point, will allow for further gene expression analyses to compare additional cartilage healing therapeutics or a transient cellular response.
ISSN:1063-4584
1522-9653
DOI:10.1016/j.joca.2015.12.001