Division of Medical Genetics, Department of Pediatrics, Duke University Medical Center, Durham, NC, USA.
Mol Genet Metab. 2013 Jun;109(2):161-70. doi: 10.1016/j.ymgme.2013.03.018. Epub 2013 Apr 6.
Glycogen Storage Disease type Ia (GSD-Ia) in humans frequently causes delayed bone maturation, decrease in final adult height, and decreased growth velocity. This study evaluates the pathogenesis of growth failure and the effect of gene therapy on growth in GSD-Ia affected dogs and mice. Here we found that homozygous G6pase (-/-) mice with GSD-Ia have normal growth hormone (GH) levels in response to hypoglycemia, decreased insulin-like growth factor (IGF) 1 levels, and attenuated weight gain following administration of GH. Expression of hepatic GH receptor and IGF 1 mRNAs and hepatic STAT5 (phospho Y694) protein levels are reduced prior to and after GH administration, indicating GH resistance. However, restoration of G6Pase expression in the liver by treatment with adeno-associated virus 8 pseudotyped vector expressing G6Pase (AAV2/8-G6Pase) corrected body weight, but failed to normalize plasma IGF 1 in G6pase (-/-) mice. Untreated G6pase (-/-) mice also demonstrated severe delay of growth plate ossification at 12 days of age; those treated with AAV2/8-G6Pase at 14 days of age demonstrated skeletal dysplasia and limb shortening when analyzed radiographically at 6 months of age, in spite of apparent metabolic correction. Moreover, gene therapy with AAV2/9-G6Pase only partially corrected growth in GSD-Ia affected dogs as detected by weight and bone measurements and serum IGF 1 concentrations were persistently low in treated dogs. We also found that heterozygous GSD-Ia carrier dogs had decreased serum IGF 1, adult body weights and bone dimensions compared to wild-type littermates. In sum, these findings suggest that growth failure in GSD-Ia results, at least in part, from hepatic GH resistance. In addition, gene therapy improved growth in addition to promoting long-term survival in dogs and mice with GSD-Ia.
糖原贮积病 Ia 型(GSD-Ia)患者常导致骨骼成熟延迟、最终成人身高降低和生长速度减慢。本研究评估了 GSD-Ia 影响犬和鼠生长失败的发病机制和基因治疗的效果。我们发现,G6pase(-/-)杂合子 GSD-Ia 小鼠在低血糖反应时 GH 水平正常,IGF-1 水平降低,GH 给药后体重增加减弱。GH 给药前后肝 GH 受体和 IGF-1 mRNA 以及肝 STAT5(磷酸化 Y694)蛋白水平的表达降低,表明 GH 抵抗。然而,用腺相关病毒 8 假型载体表达 G6pase(AAV2/8-G6Pase)治疗恢复肝 G6Pase 表达可纠正体重,但不能使 G6pase(-/-)小鼠的 IGF-1 血浆水平正常化。未经治疗的 G6pase(-/-)小鼠在 12 日龄时也表现出生长板骨化严重延迟;在 6 月龄时,用 AAV2/8-G6Pase 治疗的小鼠在放射学上表现出骨骼发育不良和肢体缩短,尽管代谢得到明显纠正。此外,用 AAV2/9-G6Pase 进行基因治疗仅部分纠正 GSD-Ia 影响犬的生长,体重和骨测量以及血清 IGF-1 浓度在治疗犬中持续较低。我们还发现杂合子 GSD-Ia 携带者犬的血清 IGF-1、成年体重和骨骼尺寸与野生型同窝仔犬相比降低。总之,这些发现表明,GSD-Ia 中的生长失败至少部分是由于肝 GH 抵抗引起的。此外,基因治疗除了促进 GSD-Ia 犬和鼠的长期存活外,还改善了生长。