Aalbers Caroline J, Bevaart Lisette, Loiler Scott, de Cortie Karin, Wright J Fraser, Mingozzi Federico, Tak Paul P, Vervoordeldonk Margriet J
Arthrogen B.V., Amsterdam, the Netherlands; Division of Clinical Immunology and Rheumatology, Academic Medical Center/University of Amsterdam, Amsterdam, the Netherlands.
Center for Cellular and Molecular Therapeutics, The Children's Hospital of Philadelphia, Pennsylvania, United States of America; Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, United States of America.
PLoS One. 2015 Jun 24;10(6):e0130612. doi: 10.1371/journal.pone.0130612. eCollection 2015.
Proof of concept for local gene therapy for the treatment of arthritis with immunomodulatory cytokine interferon beta (IFN-β) has shown promising results in animal models of rheumatoid arthritis (RA). For the treatment of RA patients, we engineered a recombinant adeno-associated serotype 5 vector (rAAV5) encoding human (h)IFN-β under control of a nuclear factor κB promoter (ART-I02).
The potency of ART-I02 in vitro as well as biodistribution in vivo in arthritic animals was evaluated to characterize the vector prior to clinical application. ART-I02 expression and bioactivity after transduction was evaluated in fibroblast-like synoviocytes (FLS) from different species. Biodistribution of the vector after local injection was assessed in a rat adjuvant arthritis model through qPCR analysis of vector DNA. In vivo imaging was used to investigate transgene expression and kinetics in a mouse collagen induced arthritis model.
Transduction of RA FLS in vitro with ART-I02 resulted in high expression levels of bioactive hIFN-β. Transduction of FLS from rhesus monkeys, rodents and rabbits with ART-I02 showed high transgene expression, and hIFN-β proved bioactive in FLS from rhesus monkeys. Transgene expression and bioactivity in RA FLS were unaltered in the presence of methotrexate. In vivo, vector biodistribution analysis in rats after intra-articular injection of ART-I02 demonstrated that the majority of vector DNA remained in the joint (>93%). In vivo imaging in mice confirmed local expression of rAAV5 in the knee joint region and demonstrated rapid detectable and sustained expression up until 7 weeks.
These data show that hIFN-β produced by RA FLS transduced with ART-I02 is bioactive and that intra-articular delivery of rAAV5 drives expression of a therapeutic transgene in the joint, with only limited biodistribution of vector DNA to other tissues, supporting progress towards a phase 1 clinical trial for the local treatment of arthritis in patients with RA.
用免疫调节细胞因子β干扰素（IFN-β）进行局部基因治疗关节炎的概念验证已在类风湿性关节炎（RA）动物模型中显示出有前景的结果。为了治疗RA患者，我们构建了一种重组腺相关血清型5载体（rAAV5），其在核因子κB启动子（ART-I02）的控制下编码人（h）IFN-β。
在临床应用前，评估了ART-I02在体外的效力以及在关节炎动物体内的生物分布，以表征该载体。在来自不同物种的成纤维细胞样滑膜细胞（FLS）中评估转导后ART-I02的表达和生物活性。通过对载体DNA的qPCR分析，在大鼠佐剂性关节炎模型中评估局部注射后载体的生物分布。在小鼠胶原诱导的关节炎模型中，使用体内成像来研究转基因表达和动力学。
用ART-I02体外转导RA FLS导致生物活性hIFN-β的高表达水平。用ART-I02转导恒河猴、啮齿动物和兔子的FLS显示出高转基因表达，并且hIFN-β在恒河猴的FLS中被证明具有生物活性。在甲氨蝶呤存在的情况下，RA FLS中的转基因表达和生物活性未改变。在体内，关节内注射ART-I02后大鼠体内的载体生物分布分析表明，大部分载体DNA保留在关节中（>93%）。小鼠体内成像证实了rAAV5在膝关节区域的局部表达，并证明了直到7周都有快速可检测到的持续表达。
这些数据表明，用ART-I02转导的RA FLS产生的hIFN-β具有生物活性，并且关节内递送rAAV5可驱动治疗性转基因在关节中的表达，载体DNA在其他组织中的生物分布有限，这支持了向RA患者关节炎局部治疗的1期临床试验迈进。
