Remodelling of human osteoarthritic cartilage by FGF-2, alone or combined with Sox9 via rAAV gene transfer
Remodelling of human osteoarthritic cartilage by FGF-2, alone or combined with Sox9 via rAAV gene transfer
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DOI：
10.1111/j.1582-4934.2008.00474.x
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发表时间：
2009-08-01
影响因子：
5.3
通讯作者：
Madry, Henning
中科院分区：
文献类型：
--
作者：
Cucchiarini, Magali;Terwilliger, Ernest F.;Madry, Henning
补偿细胞外软骨基质的损失以及抵消骨关节炎中软骨细胞表型的改变对于制定针对这种疾病的有效治疗策略至关重要。在本研究中，我们分析了应用有效的基因组合来重塑人类骨关节炎（OA）软骨的好处。我们采用有前途的重组腺相关病毒 (rAAV) 载体将促有丝分裂成纤维细胞生长因子 2 (FGF-2) 因子单独或与作为基质合成关键激活剂的转录因子 Sox9 同时递送至人类正常和 OA 关节软骨细胞。我们评估了单个（FGF-2）或组合（FGF-2/SOX9）转基因表达对体外三维培养物和原位软骨外植体中软骨细胞再生活性的影响。 FGF-2 的单次过表达增强了正常软骨细胞和 OA 软骨细胞的存活和增殖，而不会刺激增加的细胞池中的基质合成过程。当 SOX9 共过表达并伴随着蛋白聚糖和 II 型胶原产量的增加时，FGF-2 的促有丝分裂特性得以维持，表明转录因子能够抵消 FGF-2 对基质积累的影响。同样重要的是，X 型胶原蛋白（肥大标志物）的表达在候选载体治疗后急剧下降。最值得注意的是，在共同治疗的人类骨关节炎软骨中所达到的活性水平与在正常软骨中观察到的活性水平相似或更高。目前的研究结果表明，OA软骨中候选因子的联合表达可以重建正常软骨的关键特征并防止代谢稳态的病理转变。这些数据为开发通过 rAAV 治疗人类 OA 的偶联基因转移方法提供了进一步的动力。
Compensating for the loss of extracellular cartilage matrix, as well as counteracting the alterations of the chondrocyte phenotype in osteoarthritis are of key importance to develop effective therapeutic strategies against this disorder. In the present study, we analysed the benefits of applying a potent gene combination to remodel human osteoarthritic (OA) cartilage. We employed the promising recombinant adeno-associated virus (rAAV) vector to deliver the mitogenic fibroblast growth factor 2 (FGF-2) factor, alone or simultaneously with the transcription factor Sox9 as a key activator of matrix synthesis, to human normal and OA articular chondrocytes. We evaluated the effects of single (FGF-2) or combined (FGF-2/SOX9) transgene expression upon the regenerative activities of chondrocytes in three-dimensional cultures in vitro and in cartilage explants in situ. Single overexpression of FGF-2 enhanced the survival and proliferation of both normal and OA chondrocytes, without stimulating the matrix synthetic processes in the increased pools of cells. The mitogenic properties of FGF-2 were maintained when SOX9 was co-overexpressed and concomitant with an increase in the production of proteoglycans and type-II collagen, suggesting that the transcription factor was capable of counterbalancing the effects of FGF-2 on matrix accumulation. Also important, expression of type-X collagen, a marker of hypertrophy strongly decreased following treatment by the candidate vectors. Most remarkably, the levels of activities achieved in co-treated human OA cartilage were similar to or higher than those observed in normal cartilage. The present findings show that combined expression of candidate factors in OA cartilage can re-establish key features of normal cartilage and prevent the pathological shift of metabolic homeostasis. These data provide further motivation to develop coupled gene transfer approaches via rAAV for the treatment of human OA.