Successful genetic transduction in vivo into synovium by means of electroporation

Ohashi, Suzuyo; Kubo, Toshikazu; Kishida, Tsunao; Ikeda, Takumi; Takahashi, Kenji; Arai, Yuji; Terauchi, Ryu; Asada, Hidetsugu; Imanishi, Jirô; Mazda, Osam

doi:10.1016/s0006-291x(02)00386-8

Cited by 63 publications

(51 citation statements)

References 39 publications

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“…As mentioned above, in this study we focused on the functional analysis of cathepsin K as a candidate gene associated with OA pathogenesis. Gene expression in synovium can be easily manipulated, as compared with that in cartilage (19,20). Recently, it was recognized that use of siRNA makes it possible to perform specific gene-silencing experiments in vivo (21)(22)(23).…”

Section: Discussionmentioning

confidence: 99%

Down‐regulation of cathepsin K in synovium leads to progression of osteoarthritis in rabbits

Takahashi

Iwasaki

Kawa

et al. 2009

Arthritis & Rheumatism

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Objective. The hypothesis of this study was that synovial factors playing a pivotal role in the pathogenesis of osteoarthritis (OA) and thus gene expression in the synovium would be altered at the initial stage of OA. The aims of this study were to identify the candidate genes in synovium related to OA initiation, to evaluate cartilage degeneration after knockdown of the gene using small interfering RNA (siRNA) gene silencing in the knee joints at the initial stage of OA, and to determine the potential role of the knocked-down gene in OA initiation.Methods. Genes overexpressed in synovium at the initial stage of disease in a rabbit model of anterior cruciate ligament transection (ACLT)-induced OA were identified using the suppression subtractive hybridization technique and differential screening. Candidate gene expression in the synovium of the knees of rabbits with OA was manipulated with electroporation-assisted siRNA transduction 4 times before and after operation. Four weeks after surgery, histologic analysis was performed.Results. Cathepsin K gene and protein expression was significantly up-regulated in synovium at the initial stage of OA in rabbits. Down-regulation of cathepsin K in synovium at the initial stage of OA significantly accelerated cartilage degeneration.Conclusion. These results indicate that cathepsin K plays a protective role in cartilage degeneration at the initial stage of OA. We believe that the current results obtained from models of the early phase of OA will provide useful information for developing a novel strategy to prevent disease progression.

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Section: Discussionmentioning

confidence: 99%

Down‐regulation of cathepsin K in synovium leads to progression of osteoarthritis in rabbits

Takahashi

Iwasaki

Kawa

et al. 2009

Arthritis & Rheumatism

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“…Biventricular muscle was minced with a pair of scissors and homogenized in 200 ll of reporter lysis buffer (Promega, Madison, WI, USA) using a sonicator. The extract was centrifuged at 14,000g for 15 min, and the Luc activity in the supernatant was measured as described [32]. Organs other than the heart were also treated as described above.…”

Section: Methodsmentioning

confidence: 99%

Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart

Tsunoda

Mazda

Oda

et al. 2005

Biochemical and Biophysical Research Communications

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129

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Naked plasmid DNA (pDNA) and short interfering RNA (siRNA) duplexes were transduced into adult murine heart by means of sonoporation using the third-generation microbubble, BR14. Plasmid DNAs carrying luciferase, b-galactosidase (b-gal), or enhanced green fluorescent protein (EGFP) reporter genes were mixed with BR14 and injected percutaneously into the left ventricular (LV) cavity of C57BL/6 mice while exposed to transthoracic ultrasound at 1 MHz for 60 s. Sonoporation at an output intensity of 2.0 W/cm 2 and a 50% pulse duty ratio resulted in the highest luciferase expression in the heart. Histological examinations revealed significant expression of the b-gal and EGFP reporters in the subendocardial myocardium of LV. Intraventricular co-injection of siRNA-GFP and BR14 with concomitant ultrasonic exposure resulted in substantial reduction in EGFP expression in the coronary artery in EGFP transgenic mice. The present method may be applicable to gain-of-function and loss-of-function genetic engineering in vivo of adult murine heart.

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“…This technique is used in intra-articular gene therapy, but the expression of the factor is often limited to synovial membrane [11]. In addition, it is reported that gene transfer into arthritic joints by electroporation, used to deliver antiinflammatory cytokines, has a short duration of transgene expression, preventing its use for the treatment of arthritis [12].…”

Section: Non-viral Vectormentioning

confidence: 99%

Gene therapy for chondral and osteochondral regeneration: is the future now?

Bellavia

Veronesi

Carina

et al. 2017

Cell. Mol. Life Sci.

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Gene therapy might represent a promising strategy for chondral and osteochondral defects repair by balancing the management of temporary joint mechanical incompetence with altered metabolic and inflammatory homeostasis. This review analysed preclinical and clinical studies on gene therapy for the repair of articular cartilage defects performed over the last 10 years, focussing on expression vectors (non-viral and viral), type of genes delivered and gene therapy procedures (direct or indirect). Plasmids (non-viral expression vectors) and adenovirus (viral vectors) were the most employed vectors in preclinical studies. Genes delivered encoded mainly for growth factors, followed by transcription factors, anti-inflammatory cytokines and, less frequently, by cell signalling proteins, matrix proteins and receptors. Direct injection of the expression vector was used less than indirect injection of cells, with or without scaffolds, transduced with genes of interest and then implanted into the lesion site. Clinical trials (phases I, II or III) on safety, biological activity, efficacy, toxicity or bio-distribution employed adenovirus viral vectors to deliver growth factors or anti-inflammatory cytokines, for the treatment of osteoarthritis or degenerative arthritis, and tumour necrosis factor receptor or interferon for the treatment of inflammatory arthritis

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Successful genetic transduction in vivo into synovium by means of electroporation

Cited by 63 publications

References 39 publications

Down‐regulation of cathepsin K in synovium leads to progression of osteoarthritis in rabbits

Down‐regulation of cathepsin K in synovium leads to progression of osteoarthritis in rabbits

Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart

Gene therapy for chondral and osteochondral regeneration: is the future now?

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