Huang Wei, Bates Rhiannon, Appana Bhavya, Mohammed Tawfiq, Cao Lei
Department of Cancer Biology & Genetics, College of Medicine, The Ohio State University, Columbus, OH 43210, USA.
The Ohio State University Comprehensive Cancer Center, Columbus, OH 43210, USA.
iScience. 2024 Sep 17;27(10):110930. doi: 10.1016/j.isci.2024.110930. eCollection 2024 Oct 18.
AAV vectors are mainstream delivery platforms in gene therapy, yet AAV-mediated gene transfer to adipose tissue is underdeveloped due to low efficiency of natural AAVs. We previously demonstrated that an engineered capsid Rec2 displayed improved adipo-tropism but with the caveat of liver transduction. To generate highly adipo-tropic capsid, we modified Rec2 capsid by site-specific mutagenesis and found the variant V7 with F503Y, Y708D and K709I substitution to harbor highly selective adipo-tropism while diminishing liver transduction. Intraperitoneal injection favored transduction to visceral fat while intravenous administration favored subcutaneous fat. Intraperitoneal administration of V7 vector harboring human leptin and adiponectin as single transcript normalized the metabolic dysfunction of mice at a low dose. Moreover, introducing the same mutagenesis to AAV8 capsid diminished liver transduction suggesting F503, Y708 and K709 critical for liver transduction. The Rec2.V7 vector may provide a powerful tool for basic research and potent vehicle for adipose-targeting gene therapy.
腺相关病毒（AAV）载体是基因治疗中的主流递送平台，但由于天然AAV的效率较低，AAV介导的基因向脂肪组织的转移尚不完善。我们之前证明，工程化衣壳Rec2表现出改善的脂肪嗜性，但存在肝脏转导的问题。为了产生高度脂肪嗜性的衣壳，我们通过位点特异性诱变对Rec2衣壳进行了修饰，发现具有F503Y、Y708D和K709I取代的变体V7具有高度选择性的脂肪嗜性，同时减少了肝脏转导。腹腔注射有利于向内脏脂肪的转导，而静脉内给药有利于皮下脂肪。腹腔内给予携带人瘦素和脂联素作为单一转录本的V7载体，在低剂量下可使小鼠的代谢功能障碍正常化。此外，将相同的诱变引入AAV8衣壳可减少肝脏转导，表明F503、Y708和K709对肝脏转导至关重要。Rec2.V7载体可能为基础研究提供强大工具，并为脂肪靶向基因治疗提供有效的载体。
