Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania, USA.
Shriners Hospitals Pediatric Research Center, Philadelphia, Pennsylvania, USA.
Hum Gene Ther. 2023 Aug;34(15-16):682-696. doi: 10.1089/hum.2022.211.
Neurodegeneration and cerebrovascular disease share an underlying microvascular dysfunction that may be remedied by selective transgene delivery. To date, limited options exist in which cellular components of the brain vasculature can be effectively targeted by viral vector therapeutics. In this study, we characterize the first engineered adeno-associated virus (AAV) capsid mediating high transduction of cerebral vascular pericytes and smooth muscle cells (SMCs). We performed two rounds of selection with an AAV capsid scaffold displaying a heptamer peptide library to isolate capsids that traffic to the brain after intravenous delivery. One identified capsid, termed AAV-PR, demonstrated high transduction of the brain vasculature, in contrast to the parental capsid, AAV9, which transduces mainly neurons and astrocytes. Further analysis using tissue clearing, volumetric rendering, and colocalization revealed that AAV-PR enabled high transduction of cerebral pericytes located on small-caliber vessels and SMCs in the larger arterioles and penetrating pial arteries. Analysis of tissues in the periphery indicated that AAV-PR also transduced SMCs in large vessels associated with the systemic vasculature. AAV-PR was also able to transduce primary human brain pericytes with higher efficiency than AAV9. Compared with previously published AAV capsids tropisms, AAV-PR represents the first capsid to allow for effective transduction of brain pericytes and SMCs and offers the possibility of genetically modulating these cell types in the context of neurodegeneration and other neurological diseases.
神经退行性变和脑血管疾病共享潜在的微血管功能障碍，这可能通过选择性转基因传递得到纠正。迄今为止，可供选择的方法有限，其中脑血管的细胞成分可以通过病毒载体治疗有效地靶向。在这项研究中，我们描述了第一个能够介导脑血管周细胞和平滑肌细胞（SMC）高转导的工程腺相关病毒（AAV）衣壳。我们用展示七肽文库的 AAV 衣壳支架进行了两轮选择，以分离在静脉内给药后能流向大脑的衣壳。一种被称为 AAV-PR 的鉴定出的衣壳，与主要转导神经元和星形胶质细胞的亲本衣壳 AAV9 相比，对脑血管具有高转导性。使用组织清除、体积渲染和共定位的进一步分析表明，AAV-PR 能够高转导位于小血管上的脑周细胞和较大动静脉和穿透软脑膜动脉中的 SMC。对周围组织的分析表明，AAV-PR 还可以转导与全身脉管系统相关的大血管中的 SMC。AAV-PR 对原代人脑周细胞的转导效率也高于 AAV9。与以前发表的 AAV 衣壳的趋向性相比，AAV-PR 是第一个能够有效转导脑周细胞和 SMC 的衣壳，为在神经退行性变和其他神经疾病背景下对这些细胞类型进行基因调控提供了可能性。
