使用腺相关病毒（AAV）在非人灵长类动物全脑进行静脉内功能性基因转移。

Intravenous functional gene transfer throughout the brain of non-human primates using AAV.

作者信息

Chuapoco Miguel R, Flytzanis Nicholas C, Goeden Nick, Octeau J Christopher, Roxas Kristina M, Chan Ken Y, Scherrer Jon, Winchester Janet, Blackburn Roy J, Campos Lillian J, Man Kwun Nok Mimi, Sun Junqing, Chen Xinhong, Lefevre Arthur, Singh Vikram Pal, Arokiaraj Cynthia M, Shaya Timothy F, Vendemiatti Julia, Jang Min J, Mich John, Bishaw Yeme, Gore Bryan, Omstead Victoria, Taskin Naz, Weed Natalie, Ting Jonathan, Miller Cory T, Deverman Benjamin E, Pickel James, Tian Lin, Fox Andrew S, Gradinaru Viviana

机构信息

Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.

Capsida Biotherapeutics, Thousand Oaks, CA 91320, USA.

出版信息

Res Sq. 2023 Jan 13:rs.3.rs-1370972. doi: 10.21203/rs.3.rs-1370972/v1.

DOI:10.21203/rs.3.rs-1370972/v1

PMID:36789432

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9928057/

Abstract

Adeno-associated viruses (AAVs) promise robust gene delivery to the brain through non-invasive, intravenous delivery. However, unlike in rodents, few neurotropic AAVs efficiently cross the blood-brain barrier in non-human primates (NHPs). Here we describe AAV.CAP-Mac, an engineered variant identified by screening in adult marmosets and newborn macaques with improved efficiency in the brain of multiple NHP species: marmoset, rhesus macaque, and green monkey. CAP-Mac is neuron-biased in infant Old World primates, exhibits broad tropism in adult rhesus macaques, and is vasculature-biased in adult marmosets. We demonstrate applications of a single, intravenous dose of CAP-Mac to deliver (1) functional GCaMP for ex vivo calcium imaging across multiple brain areas, and (2) a cocktail of fluorescent reporters for Brainbow-like labeling throughout the macaque brain, circumventing the need for germline manipulations in Old World primates. Given its capabilities for systemic gene transfer in NHPs, CAP-Mac promises to help unlock non-invasive access to the brain.

摘要

腺相关病毒（AAV）有望通过非侵入性静脉注射将基因有效递送至大脑。然而，与啮齿动物不同，在非人灵长类动物（NHP）中，很少有嗜神经AAV能有效穿过血脑屏障。在此，我们描述了AAV.CAP-Mac，这是一种通过在成年狨猴和新生猕猴中筛选鉴定出的工程变体，在多种NHP物种（狨猴、恒河猴和绿猴）的大脑中具有更高的效率。CAP-Mac在旧世界猴幼体中偏向神经元，在成年恒河猴中表现出广泛的嗜性，在成年狨猴中偏向血管。我们展示了单次静脉注射CAP-Mac的应用，可用于（1）递送功能性GCaMP用于多个脑区的离体钙成像，以及（2）递送一组荧光报告基因用于猕猴全脑的类Brainbow标记，从而避免了对旧世界猴进行种系操作的需要。鉴于其在NHP中进行全身基因转移的能力，CAP-Mac有望帮助实现对大脑的非侵入性访问。