Comparative Efficacy and Safety of Multiple Routes of Direct CNS Administration of Adeno-Associated Virus Gene Transfer Vector Serotype rh.10 Expressing the Human Arylsulfatase A cDNA to Nonhuman Primates
Comparative Efficacy and Safety of Multiple Routes of Direct CNS Administration of Adeno-Associated Virus Gene Transfer Vector Serotype rh.10 Expressing the Human Arylsulfatase A cDNA to Nonhuman Primates
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DOI：
10.1089/humc.2013.239
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发表时间：
2014-09-01
影响因子：
--
通讯作者：
Crystal, Ronald G.
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文献类型：
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作者：
Rosenberg, Jonathan B.;Sondhi, Dolan;Crystal, Ronald G.
异染性脑白质营养不良（MLD）是一种由溶酶体酶芳基硫酸酯酶A（ARSA）缺乏引起的致命疾病，与硫苷积累有关，导致中枢和外周神经系统广泛脱髓鞘。基于先前的研究表明腺相关病毒AAVrh.10可以介导分泌的溶酶体转基因在CNS中的广泛分布，并且作为人体试验的前奏，我们在大型动物模型中比较评估了编码人ARSA的AAVrh.10载体的最佳CNS递送途径，以获得ARSA酶的最广泛分布。测试了五条路线（每个总剂量，1.5 × 10（12）个基因组拷贝的AAVrh. 10 hARSA-FLAG）：（1）递送至卵圆中心的白色物质;（2）递送至深部灰质（壳核，丘脑和尾状核）加上覆盖的白色物质;（3）对流增强输送到相同的深部灰质位置;（4）侧脑室;和（5）动脉内递送高渗甘露醇至大脑中动脉。13周后，三种直接脑实质内给药途径的ARSA活性分布均显著高于磷酸盐缓冲盐水给药对照组，但脑室内和动脉内给药途径未能显示出高于对照组的可测量水平。免疫组织化学染色在皮质，白色物质，深灰质的纹状体，丘脑，脉络丛，脊髓背根神经节证实了这些结果。在研究的五种途径中，给予白色物质产生了最广泛的ARSA分布，80%的大脑显示出高于PBS对照的ARSA活性的治疗性（10%）增加。通过安全性参数测量，任何给药途径均未观察到显著毒性，但通过组织病理学观察到一些炎症变化。我们得出结论，AAVrh.10介导的ARSA通过CNS给药递送到白色物质中可能是安全的，并产生最广泛的ARSA分布，使其成为最合适的载体递送途径。
Metachromatic leukodystrophy (MLD), a fatal disorder caused by deficiency of the lysosomal enzyme arylsulfatase A (ARSA), is associated with an accumulation of sulfatides, causing widespread demyelination in both central and peripheral nervous systems. On the basis of prior studies demonstrating that adeno-associated virus AAVrh.10 can mediate widespread distribution in the CNS of a secreted lysosomal transgene, and as a prelude to human trials, we comparatively assessed the optimal CNS delivery route of an AAVrh.10 vector encoding human ARSA in a large animal model for broadest distribution of ARSA enzyme. Five routes were tested (each total dose, 1.5x10(12) genome copies of AAVrh.10hARSA-FLAG): (1) delivery to white matter centrum ovale; (2) deep gray matter delivery (putamen, thalamus, and caudate) plus overlying white matter; (3) convection-enhanced delivery to same deep gray matter locations; (4) lateral cerebral ventricle; and (5) intraarterial delivery with hyperosmotic mannitol to the middle cerebral artery. After 13 weeks, the distribution of ARSA activity subsequent to each of the three direct intraparenchymal administration routes was significantly higher than in phosphate-buffered saline-administered controls, but administration by the intraventricular and intraarterial routes failed to demonstrate measurable levels above controls. Immunohistochemical staining in the cortex, white matter, deep gray matter of the striatum, thalamus, choroid plexus, and spinal cord dorsal root ganglions confirmed these results. Of the five routes studied, administration to the white matter generated the broadest distribution of ARSA, with 80% of the brain displaying more than a therapeutic (10%) increase in ARSA activity above PBS controls. No significant toxicity was observed with any delivery route as measured by safety parameters, although some inflammatory changes were seen by histopathology. We conclude that AAVrh.10-mediated delivery of ARSA via CNS administration into the white matter is likely to be safe and yields the widest distribution of ARSA, making it the most suitable route of vector delivery.