Optimization of gene transfer conditions to adipose tissue and application toward hemophilia

脂肪组织可能是血友病等疾病基因转移的合适目标。然而，目前尚无有效的体内基因转移方法。在这项研究中，我们探索了辅料在增加基因转移到小鼠脂肪组织中的作用，并发现Pluronic F88在与AAV血清型1载体结合时可用于此目的。编码小鼠促红细胞生成素的载体也证明了这种改善，小鼠变得红细胞增多。此外，在去除转导脂肪组织后，血浆促红细胞生成素水平恢复正常，这表明该方法的独特优势。以同样的方式检测基于aav8的载体时，在肝脏而不是注射的脂肪组织中观察到稳健的转基因表达。这一发现导致肌肉注射基于aav8的载体后发现异位肝脏表达。另一方面，在用载体编码凝血因子IX基因的实验中，小鼠的转基因表达水平不理想。造成这种现象的原因尚不清楚，需要在未来澄清。为了提高转基因基因在血友病以外疾病中的表达水平和应用，有必要进一步优化和分析。

Adipose tissue is potentially a suitable target of gene transfer for diseases like hemophilia. However, no efficient method has been developed for in vivo gene transfer. In this study, we explored the utility of excipients to augment gene transfer into the adipose tissue of mice and found that Pluronic F88 was useful for this purpose when combined with AAV serotype 1 vectors. The improvement was also demonstrated by vectors encoding murine erythropoietin, and the mice became polycythemic. Moreover, after removing transduced adipose tissue, plasma erythropoietin levels returned to normal, which suggests the unique advantage of this method. When AAV8-based vector was tested in the same manner, robust transgene expression was observed in the liver rather than the injected adipose tissue. This finding led to the discovery of ectopic liver expression following intramuscular injection of AAV8-based vectors. On the other hand, in experiments with vectors encoding coagulation factor IX gene, transgene expression level was not satisfactory in mice. The reason for this phenomenon is still unclear and need to be clarified in the future. Further optimization and analysis is necessary to improve the level of transgene expression and application to the diseases other than hemophilia.