Yadav Alka, Liang Rich, Press Kelly, Schmidt Annika, Shabani Zahra, Leng Kun, Wang Calvin, Sekhar Abinav, Shi Joshua, Devlin Garth W, Gonzalez Trevor J, Asokan Aravind, Su Hua
University of California, San Francisco.
Duke University School of Medicine.
Res Sq. 2024 Jun 12:rs.3.rs-4469011. doi: 10.21203/rs.3.rs-4469011/v1.
Nosebleeds and intracranial hemorrhage from brain arteriovenous malformations (bAVMs) are among the most devastating symptoms of patients with hereditary hemorrhagic telangiectasis (HHT). All available managements have limitations. We showed that intravenous delivery of soluble FMS-related tyrosine kinase 1 using an adeno-associated viral vector (AAV9-sFLT1) reduced bAVM severity of deficient mice. However, minor liver inflammation and growth arrest in young mice were observed. To identify AAV variants and delivery methods that can best transduce brain and nasal tissue with an optimal transduction profile, we compared 3 engineered AAV capsids (AAV.cc47, AAV.cc84 and AAV1RX) with AAV9. A single-stranded CBA promoter driven tdTomato transgene was packaged in these capsids and delivered intravenously (i.v.) or intranasally (i.n.) to wild-type mice. A CMV promoter driven transgene was packaged into AAV.cc84 and delivered to PdgfbiCre; mice through i.v. injection followed by brain AVM induction. Transduced cells in different organs, vessel density and abnormal vessels in the bAVMs, and liver inflammation were analyzed histologically. Liver and kidney function were measured enzymatically. Compared to other viral vectors, AAV.cc84, after i.v. delivery, transduced a high percentage of brain ECs and few hepatocytes; whereas after i.n. delivery, AAV.cc84 transduced ECs and perivascular cells in the brain, and ECs, epithelial cells, and skeletal muscles in the nose with minimum hepatocyte transduction. No changes to liver or kidney function were detected. Delivery of AAV.cc84-Alk1 through i.v. to PdgfbiCre; mice reduced bAVM severity. In summary, we propose that AAV.cc84-Alk1 is a promising candidate for developing gene therapy in HHT patients.
鼻出血和脑动静脉畸形（bAVM）导致的颅内出血是遗传性出血性毛细血管扩张症（HHT）患者最具毁灭性的症状之一。所有可用的治疗方法都有局限性。我们发现，使用腺相关病毒载体静脉注射可溶性FMS相关酪氨酸激酶1（AAV9-sFLT1）可降低缺陷小鼠的bAVM严重程度。然而，在幼鼠中观察到了轻微的肝脏炎症和生长停滞。为了确定能够以最佳转导谱最佳转导脑和鼻组织的腺相关病毒变体和递送方法，我们将3种工程化腺相关病毒衣壳（AAV.cc47、AAV.cc84和AAV1RX）与AAV9进行了比较。将单链CBA启动子驱动的tdTomato转基因包装在这些衣壳中，并通过静脉内（i.v.）或鼻内（i.n.）递送至野生型小鼠。将CMV启动子驱动的转基因包装到AAV.cc84中，并通过静脉注射递送至PdgfbiCre;小鼠，随后诱导脑AVM。通过组织学分析不同器官中的转导细胞、bAVM中的血管密度和异常血管以及肝脏炎症。通过酶法测量肝功能和肾功能。与其他病毒载体相比，静脉注射后，AAV.cc84转导了高比例的脑内皮细胞（ECs）和少量肝细胞；而鼻内递送后，AAV.cc84转导了脑中的ECs和血管周围细胞，以及鼻中的ECs、上皮细胞和骨骼肌，肝细胞转导最少。未检测到肝功能或肾功能的变化。通过静脉注射将AAV.cc84-Alk1递送至PdgfbiCre;小鼠可降低bAVM严重程度。总之，我们认为AAV.cc84-Alk1是开发HHT患者基因治疗的有前景的候选者。
