Gene therapy for familial amyloidotic polyneuropathy (FAP)

肝移植治疗家族性淀粉样多发性神经病（FAP）的研究表明，替换甲状腺素运载蛋白（TTR）变异基因可有效地终止FAP的临床症状。因此，我们需要开发一种新的治疗方法，防止在肝脏和视网膜中产生TTR变体。在这项研究中，我们使用HepG2细胞显示在体外转换TTR基因的单链寡核苷酸（SSO），嵌在去端胶原，旨在促进内源性修复的基因组DNA。对于研究的体内部分，我们使用来自转基因小鼠的兔眼和肝脏，所述转基因小鼠的固有野生型TTR基因被鼠TTR Val 30Met基因替换。基因转换率通过实时RCR结合等位基因特异性扩增来确定。我们的研究结果表明，在HepG2细胞，兔眼和转基因小鼠的肝脏中，基因转换率分别约为总TTR基因的11%，1%和9%。因此，通过这种方法的基因治疗可能是一个有前途的替代肝移植治疗FAP。

Liver transplantation for familial amyloidotic polyneuropathy (FAP) revealed that replacement of the variant transthyretin (TTR) gene is effective for halting clinical symptoms of FAP. Thus, we need to develop a new treatment that prevents production of the variant TTR in the liver and retina. In this study, we used HepG2 cells to show in vitro conversion of the TTR gene by single-stranded oligonucleotides (SSOs), embedded in atelocollagen, designed to promote endogenous repair of genomic DNA. For the in vivo portion of the study, we used rabbit eyes and liver from transgenic mice whose intrinsic wild-type TTR gene was replaced by the murine TTR Val30Met gene. The rate of gene conversion was determined by real-time RCR combined with mutual-allele-specific amplification. Our results indicate that the rate of gene conversion was approximately 11%, 1%, and 9% of the total TTR gene in HepG2 cells, rabbit eyes, and liver from transgenic mice, respectively. Gene therapy via this method may therefore be a promising alternative to liver transplantation for treatment of FAP.