Marottoli Felecia M, Balu Deebika, Chaudhary Rohan, Lutz Sarah E, Tai Leon M
Department of Anatomy and Cell Biology, University of Illinois at Chicago, Chicago, IL, USA.
ASN Neuro. 2024;16(1):2427953. doi: 10.1080/17590914.2024.2427953. Epub 2024 Dec 2.
Brain endothelial cells are critical for homeostasis of the central nervous system. Novel adeno-associated viruses (AAV) with brain endothelial cell tropism have been developed and are beginning to be employed in mechanistic and therapeutic research. Studies using AAVs can be involved in terms of cost, time and personnel, and many groups, including our own, are not experts on the technology. Therefore, it is important to report data using AAVs with the research community as a guide for ongoing and future studies. Here, we detail our initial experience with the two most prevalent AAVs with tropism for brain endothelial cells, AAV-BR1 and AAV-BI30. One of our long-term goals is to express key proteins in brain endothelial cells and determine the impact on brain function. For method development, we administered AAV-BR1 and AAV-BI30 with a CMV-driven fluorescent reporter (CMV-P2A-mCherry) to wild-type mice intravenously (retro-orbital) and measured expression in brain and peripheral tissues by RT-PCR and immunostaining. We found that AAV-BR1 transduces neurons and endothelial cells in the brain, and the lung and liver, whereas AAV-BI30 transduces brain endothelial cells and peripheral tissue. Our data highlights the importance of using the AAV best suited to the scientific question.
脑内皮细胞对中枢神经系统的稳态至关重要。具有脑内皮细胞嗜性的新型腺相关病毒(AAV)已被开发出来,并开始用于机制研究和治疗研究。使用AAV的研究在成本、时间和人员方面都可能涉及,包括我们自己在内的许多团队都不是该技术的专家。因此,以研究界为指导,报告使用AAV的数据对于正在进行的和未来的研究很重要。在这里,我们详细介绍了我们对两种最常见的具有脑内皮细胞嗜性的AAV,即AAV-BR1和AAV-BI30的初步经验。我们的一个长期目标是在脑内皮细胞中表达关键蛋白,并确定其对脑功能的影响。为了进行方法开发,我们将携带巨细胞病毒(CMV)驱动的荧光报告基因(CMV-P2A-mCherry)的AAV-BR1和AAV-BI30经静脉(眶后)注射到野生型小鼠体内,并通过逆转录聚合酶链反应(RT-PCR)和免疫染色测量脑和外周组织中的表达。我们发现,AAV-BR1可转导脑中的神经元和内皮细胞以及肺和肝脏中的细胞,而AAV-BI30可转导脑内皮细胞和外周组织。我们的数据突出了使用最适合科学问题的AAV的重要性。