Ciron Carine, Cressant Arnaud, Roux Françoise, Raoul Sylvie, Cherel Yan, Hantraye Philippe, Déglon Nicole, Schwartz Bertrand, Barkats Martine, Heard Jean-Michel, Tardieu Marc, Moullier Philippe, Colle Marie-Anne
INSERM U649, 44000 Nantes, France.
Hum Gene Ther. 2009 Apr;20(4):350-60. doi: 10.1089/hum.2008.155.
We have previously demonstrated that delivery of a recombinant adeno-associated virus (rAAV) encoding human alpha-iduronidase (hIDUA) in the putamen and centrum semiovale was feasible and beneficial in a dog model of Hurler's syndrome. In the present study, we investigated the safety and vector diffusion profile of three rAAV serotypes (rAAV2/1, rAAV2/2, and rAAV2/5), encoding hIDUA in the central and peripheral nervous systems of nonhuman primates. Six macaques received the same vector dose injected into the right putamen and the homolateral internal capsule. Neurological examinations were done regularly and showed no detectable clinical consequence of the intracerebral injections. Because transgene IDUA was indistinguishable from endogenous enzymatic activity, we looked for vector diffusion by performing quantitative polymerase chain reaction on serial sections from the brain and spinal cord. We found that global diffusion throughout the brain was not significantly different between the three serotypes. However, rAAV2/1 and rAAV2/5 resulted in higher vector copy numbers per cell than did rAAV2/2, respectively, in the brain and the distal neuronal structures (spinal cord and peripheral nerves).
我们之前已经证明,在胡尔勒综合征犬模型中,将编码人α-艾杜糖醛酸酶(hIDUA)的重组腺相关病毒(rAAV)注射到壳核和半卵圆中心是可行且有益的。在本研究中,我们调查了三种rAAV血清型(rAAV2/1、rAAV2/2和rAAV2/5)在非人灵长类动物中枢和外周神经系统中编码hIDUA时的安全性和载体扩散情况。六只猕猴接受了相同载体剂量的注射,注射部位为右侧壳核和同侧内囊。定期进行神经学检查,结果显示脑内注射未产生可检测到的临床后果。由于转基因IDUA与内源性酶活性无法区分,我们通过对脑和脊髓的连续切片进行定量聚合酶链反应来寻找载体扩散情况。我们发现,三种血清型在全脑的扩散情况没有显著差异。然而,在脑和远端神经结构(脊髓和外周神经)中,rAAV2/1和rAAV2/5分别比rAAV2/2产生了更高的每细胞载体拷贝数。