Gene therapy for familial amyloidotic polyneuropathy (FAP)

家族性淀粉样变性多神经病变（FAP）的肝移植研究表明，置换变异型甲状腺素（TTR）基因可有效阻止FAP的临床症状。因此，我们需要开发一种新的治疗方法来阻止肝脏和视网膜中TTR变体的产生。在这项研究中，我们使用HepG2细胞，展示了通过嵌入胶原内的单链寡核苷酸（SSOs）在体外转化TTR基因，旨在促进基因组DNA的内源性修复。在体内部分的研究中，我们使用了兔眼和转基因小鼠的肝脏，这些小鼠的内在野生型TTR基因被小鼠TTR Val30Met基因所取代。采用实时RCR结合互等位基因特异性扩增测定基因转化率。我们的研究结果表明，在HepG2细胞、兔眼和转基因小鼠肝脏中，基因转化率分别约为总TTR基因的11%、1%和9%。因此，通过这种方法进行基因治疗可能是肝移植治疗FAP的一种有希望的替代方法。

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.