In Vitro and in Vivo Testing of a Novel Regulatory System for Gene Therapy for Intervertebral Disc Degeneration

Sowa, Gwendolyn; Westrick, Edward; Pacek, Corey A.; Coelho, Paulo; Patel, Devin; Vadalà, Gianluca; Georgescu, Helga I.; Vo, Nam; Studer, Rebecca K.; Kang, James D.

doi:10.1097/brs.0b013e3181ed11c1

Cited by 29 publications

(17 citation statements)

References 8 publications

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“…As such, with the removal of the induction agent, the protein production of the introduced gene ceases. The ability to control temporal expression of the therapeutic gene also allows for control and safe administration of these potentially powerful compounds [86].…”

Section: Gene-based Therapiesmentioning

confidence: 99%

Molecular Basis of Intervertebral Disc Degeneration and Herniations: What Are the Important Translational Questions?

Kadow

Sowa

et al. 2015

Clinical Orthopaedics &Amp; Related Research

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Background Intervertebral disc degeneration is a common condition with few inexpensive and effective modes of treatment, but current investigations seek to clarify the underlying process and offer new treatment options. It will be important for physicians to understand the molecular basis for the pathology and how it translates to developing clinical treatments for disc degeneration. In this review, we sought to summarize for clinicians what is known about the molecular processes that causes disc degeneration.

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Section: Gene-based Therapiesmentioning

confidence: 99%

Molecular Basis of Intervertebral Disc Degeneration and Herniations: What Are the Important Translational Questions?

Kadow

Sowa

et al. 2015

Clinical Orthopaedics &Amp; Related Research

Self Cite

217

197

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“…This quantity was based on previously published dosing [14]. Prior studies proved that the small diameter needle and the effect of injecting a small volume of saline [23], or a nontherapeutic gene [24], do not induce disc degeneration.…”

Section: Sample Preparationmentioning

confidence: 99%

Injection of AAV2-BMP2 and AAV2-TIMP1 into the nucleus pulposus slows the course of intervertebral disc degeneration in an in vivo rabbit model

et al. 2012

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BACKGROUND CONTEXT Intervertebral disc degeneration (IDD) is a common cause of back pain. Patients who fail conservative management may face the morbidity of surgery. Alternative treatment modalities could have a significant impact on disease progression and patients’ quality of life. PURPOSE To determine if the injection of a virus vector carrying a therapeutic gene directly into the nucleus pulposus improves the course of IDD. STUDY DESIGN Prospective randomized controlled animal study. METHODS Thirty-four skeletally mature New Zealand white rabbits were used. In the treatment group, L2–L3, L3–L4, and L4–L5 discs were punctured in accordance with a previously validated rabbit annulotomy model for IDD and then subsequently treated with adeno-associated virus serotype 2 (AAV2) vector carrying genes for either bone morphogenetic protein 2 (BMP2) or tissue inhibitor of metalloproteinase 1 (TIMP1). A nonoperative control group, nonpunctured sham surgical group, and punctured control group were also evaluated. Serial magnetic resonance imaging (MRI) studies at 0, 6, and 12 weeks were obtained, and a validated MRI analysis program was used to quantify degeneration. The rabbits were sacrificed at 12 weeks, and L4–L5 discs were analyzed histologically. Viscoelastic properties of the L3–L4 discs were analyzed using uniaxial load normalized displacement testing. Creep curves were mathematically modeled according to a previously validated two-phase exponential model. Serum samples obtained at 0, 6, and 12 weeks were assayed for biochemical evidence of degeneration. RESULTS The punctured group demonstrated MRI and histologic evidence of degeneration as expected. The treatment groups demonstrated less MRI and histologic evidence of degeneration than the punctured group. The serum biochemical marker C-telopeptide of collagen type II increased rapidly in the punctured group, but the treated groups returned to control values by 12 weeks. The treatment groups demonstrated several viscoelastic properties that were distinct from control and punctured values. CONCLUSIONS Treatment of punctured rabbit intervertebral discs with AAV2-BMP2 or AAV2-TIMP1 helps delay degenerative changes, as seen on MRI, histologic sampling, serum biochemical analysis, and biomechanical testing. Although data from animal models should be extrapolated to the human condition with caution, this study supports the potential use of gene therapy for the treatment of IDD.

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“…Molecular therapies, including the use of growth factors (26), inflammation inhibitors (27), and proteinase inhibitors (28), have exhibited limited therapeutic durations and are not suitable for treating chronic degeneration processes. Gene therapies, using virus vectors or plasmids encoding exogenous proteins to stimulate matrix synthesis or inhibit its degradation, have overcome the limitations of molecular treatment (24,29-31). Cell-based therapies, including reimplantation of nucleus pulposus cells or stem cells, also have shown exciting results in animal experiments (24,30,32,33).…”

Section: Discussionmentioning

confidence: 99%

Lentiviral-mediated RNAi targeting caspase-3 inhibits apoptosis induced by serum deprivation in rat endplate chondrocytes in vitro

Ding

et al. 2014

Braz J Med Biol Res

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Current studies find that degenerated cartilage endplates (CEP) of vertebrae, with fewer diffusion areas, decrease nutrient supply and accelerate intervertebral disc degeneration. Many more apoptotic cells have been identified in degenerated than in normal endplates, and may be responsible for the degenerated grade. Previous findings suggest that inhibition of apoptosis is one possible approach to improve disc regeneration. It is postulated that inhibition of CEP cell apoptosis may be responsible for the regeneration of endplates. Caspase-3, involved in the execution phase of apoptosis, is a candidate for regulating the apoptotic process. In the present study, CEP cells were incubated in 1% fetal bovine serum. Activated caspases were detected to identify the apoptotic pathway, and apoptosis was quantified by flow cytometry. Lentiviral caspase-3 short hairpin RNA (shRNA) was employed to study its protective effects against serum deprivation. Silencing of caspase-3 expression was quantified by reverse transcription-polymerase chain reaction and Western blots, and inhibition of apoptosis was quantified by flow cytometry. Serum deprivation increased apoptosis of rat CEP cells through activation of a caspase cascade. Lentiviral caspase-3 shRNA was successfully transduced into CEP cells, and specifically silenced endogenous caspase-3 expression. Surviving cells were protected by the downregulation of caspase-3 expression and activation. Thus, lentiviral caspase-3 shRNA-mediated RNAi successfully silenced endogenous caspase-3 expression, preventing inappropriate or premature apoptosis.

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In Vitro and in Vivo Testing of a Novel Regulatory System for Gene Therapy for Intervertebral Disc Degeneration

Cited by 29 publications

References 8 publications

Molecular Basis of Intervertebral Disc Degeneration and Herniations: What Are the Important Translational Questions?

Molecular Basis of Intervertebral Disc Degeneration and Herniations: What Are the Important Translational Questions?

Injection of AAV2-BMP2 and AAV2-TIMP1 into the nucleus pulposus slows the course of intervertebral disc degeneration in an in vivo rabbit model

Lentiviral-mediated RNAi targeting caspase-3 inhibits apoptosis induced by serum deprivation in rat endplate chondrocytes in vitro

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