UMR1195 Inserm and University Paris Saclay, Le Kremlin-Bicêtre, France.
MIRCen Institute, CEA, Fontenay-aux-Roses, France.
Hum Gene Ther. 2022 Sep;33(17-18):936-949. doi: 10.1089/hum.2021.293. Epub 2022 Jul 18.
Adrenomyeloneuropathy (AMN) is a late-onset axonopathy of spinal cord tracts caused by mutations of the ABCD1 gene that encodes adrenoleukodystrophy protein (ALDP), a peroxisomal transporter of very long-chain fatty acids (VLCFA). Disturbed metabolic interaction between oligodendrocytes (OL) and axons is suspected to play a major role in AMN axonopathy. To develop a vector targeting OL, the human ABCD1 gene driven by a short 0.3 kb part of the human myelin-associated glycoprotein (MAG) promoter was packaged into an adeno-associated viral serotype 9 (rAAV9). An intravenous injection of this vector on postnatal day 10 in Abcd1 mice, a model of AMN, allowed a near normal motor performance to persist for 24 months, while age-matched untreated mice developed major defects of balance and motricity. Three weeks postvector, 50-54% of spinal cord white matter OL was expressing human ALDP (hALDP) at the cervical level, and only 6-7% after 24 months. In addition, 29-32% of cervical spinal cord astrocytes at 3 weeks and 16-19% at 24 months also expressed ALDP. C26:0-lysoPC, a sensitive VLCFA marker of AMN, was lower by 41% and 50%, respectively, in the spinal cord and brain of vector-treated compared with untreated mice. In a nonhuman primate, the intrathecal injection of the rAAV9-MAG vector induced abundant ALDP expression at 3 weeks in spinal cord OL (43%, 29%, and 26% at cervical, thoracic, and lumbar levels) and cerebellum OL (35%). In addition, 33-41% of spinal cord astrocytes expressed hALDP, and 27% of cerebellar astrocytes. To our knowledge, OL targeting had not been obtained before in primates with other vectors or promoters. The current results thus provide a robust proof-of-concept not only for the gene therapy of AMN but also for other central nervous system diseases, where the targeting of OL with the rAAV9-MAG vector may be of interest.
肾上腺脑白质营养不良性神经病（adrenomyeloneuropathy，AMN）是一种由 ABCD1 基因突变引起的脊髓束轴突病，该基因突变编码肾上腺白质营养不良蛋白（adrenoleukodystrophy protein，ALDP），是一种过氧化物酶体长链脂肪酸（very long-chain fatty acids，VLCFA）转运体。人们怀疑少突胶质细胞（oligodendrocyte，OL）与轴突之间代谢相互作用的紊乱在 AMN 轴突病中起主要作用。为了开发针对 OL 的载体，我们将人类 ABCD1 基因与人类髓鞘相关糖蛋白（myelin-associated glycoprotein，MAG）启动子的 0.3kb 短片段包装到腺相关病毒血清型 9（adeno-associated viral serotype 9，rAAV9）中。在 AMN 模型 Abcd1 小鼠出生后 10 天静脉注射该载体，可使运动功能接近正常持续 24 个月，而年龄匹配的未治疗小鼠则出现严重的平衡和运动功能缺陷。载体注射后 3 周，在颈椎水平，50-54%的脊髓白质 OL 表达人 ALDP（human ALDP，hALDP），24 个月后仅为 6-7%。此外，在 3 周和 24 个月时，32%的颈椎脊髓星形胶质细胞和 16-19%的颈椎脊髓星形胶质细胞也表达 ALDP。C26:0-溶血磷脂酰胆碱（lysoPC）是 AMN 的一种敏感 VLCFA 标志物，与未治疗的小鼠相比，载体治疗的小鼠脊髓和大脑中的 C26:0-lysoPC 分别降低了 41%和 50%。在非人类灵长类动物中，鞘内注射 rAAV9-MAG 载体可在 3 周时在脊髓 OL（颈椎、胸和腰水平分别为 43%、29%和 26%）和小脑 OL（35%）中诱导大量 ALDP 表达。此外，33-41%的脊髓星形胶质细胞表达 hALDP，27%的小脑星形胶质细胞表达 hALDP。据我们所知，以前使用其他载体或启动子在灵长类动物中没有进行过 OL 靶向治疗。目前的结果不仅为 AMN 的基因治疗提供了强有力的概念验证，也为其他中枢神经系统疾病提供了有力的概念验证，其中 rAAV9-MAG 载体对 OL 的靶向治疗可能具有重要意义。
