Lewin A S, Drenser K A, Hauswirth W W, Nishikawa S, Yasumura D, Flannery J G, LaVail M M
Department of Molecular Genetics and Microbiology, Gene Therapy Center, University of Florida College of Medicine, Gainesville 32610, USA.
Nat Med. 1998 Aug;4(8):967-71. doi: 10.1038/nm0898-967.
Ribozymes, catalytic RNA molecules that cleave a complementary mRNA sequence, have potential as therapeutics for dominantly inherited disease. Twelve percent of American patients with the blinding disease autosomal dominant retinitis pigmentosa (ADRP) carry a substitution of histidine for proline at codon 23 (P23H) in their rhodopsin gene, resulting in photoreceptor cell death from the synthesis of the abnormal gene product. Ribozymes can discriminate and catalyze the in vitro destruction of P23H mutant mRNAs from a transgenic rat model of ADRP. Here, we demonstrate that in vivo expression of either a hammerhead or hairpin ribozyme in this rat model considerably slows the rate of photoreceptor degeneration for at least three months. Catalytically inactive control ribozymes had less effect on the retinal degeneration. Intracellular production of ribozymes in photoreceptors was achieved by transduction with a recombinant adeno-associated virus (rAAV) incorporating a rod opsin promoter. Ribozyme-directed cleavage of mutant mRNAs, therefore, may be an effective therapy for ADRP and also may be applicable to other inherited diseases.
核酶是一种能切割互补mRNA序列的催化性RNA分子，具有作为显性遗传病治疗手段的潜力。在美国，12%患有致盲性疾病常染色体显性遗传性视网膜色素变性（ADRP）的患者，其视紫红质基因的第23密码子（P23H）处存在脯氨酸被组氨酸替代的情况，这导致光感受器细胞因异常基因产物的合成而死亡。核酶能够识别并在体外催化破坏来自ADRP转基因大鼠模型的P23H突变mRNA。在此，我们证明在该大鼠模型中，锤头状或发夹状核酶的体内表达可使光感受器变性速率显著减缓至少三个月。催化失活的对照核酶对视网膜变性的影响较小。通过用携带视杆视蛋白启动子的重组腺相关病毒（rAAV）进行转导，实现了光感受器内核酶的胞内产生。因此，核酶介导的突变mRNA切割可能是治疗ADRP的有效方法，也可能适用于其他遗传性疾病。
