AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset

Goertsen, David; Flytzanis, Nicholas C.; Goeden, Nick; Chuapoco, Miguel; Cummins, Alexander; Chen, Yijing; Fan, Yingying; Zhang, Qiangge; Sharma, Jitendra; Duan, Yangyang; Wang, Liping; Feng, Guoping; Chen, Yu; Ip, Nancy Y.; Pickel, James; Gradinaru, Viviana

doi:10.1038/s41593-021-00969-4

Cited by 210 publications

(228 citation statements)

References 61 publications

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“…In the liver, variant barcodes were variably enriched relative to AAV9. Notably, CAP-Mac and CAP-C2 were negatively enriched in the liver, along with some of the previously engineered controls shown to be de-targeted from the liver in rodents 17 (Fig. 2c).…”

Section: Cap-mac Is Enriched In the Infant Rhesus Macaque Cns Compared To Other Engineered Variantsmentioning

confidence: 69%

“…In vivo AAV capsid selections have been primarily conducted in mice, partially due to the utility of Cre-transgenic mouse lines to increase selective pressure during selections, as these Cre-based selections can yield neurotropic capsids in as few as two rounds of selection 14,16,17 . However, these engineered variants have thus far failed to translate to NHPs 24,25 .…”

Section: Discussionmentioning

confidence: 99%

“…By pooling several variants into equimolar vector genome doses and administering them to the same rhesus macaque, we could limit the number of animals used for characterization while also assessing variant performance head-to-head, as we have shown that inter-animal variability exceeds intra-animal variability 51 . We pooled a total of 8 capsid variants: CAP-Mac and CAP-C2 as well as the parent capsid, AAV9, plus five other previously engineered AAV controls 15,17 Each variant packaged a single stranded human frataxin transgene fused to a hemagglutinin (HA) epitope tag under control of the ubiquitous CAG promoter (ssCAG-hFXN-HA) with a unique 12 bp molecular barcode in the 3' UTR. This construct design allowed us to assess protein expression and localization of the virus pool by staining for the HA tag.…”

Section: Cap-mac Is Enriched In the Infant Rhesus Macaque Cns Compared To Other Engineered Variantsmentioning

confidence: 99%

“…The AAV9 variant AAV-PHP.B was the first variant to unlock efficient widespread gene transfer in adult mammals, traversing the BBB after systemic intravenous (IV) administration in mice 14 . Other variants have since followed with similar or enhanced properties, such as the ability to cross the BBB across different mouse strains, decreased transduction in non-CNS tissue, and biased tropism towards cell-types in the brain [15][16][17]22,23 . While two groups reported that the BBB-crossing tropism of AAV-PHP.B does not translate to the rhesus macaque 24,25 , studies reporting the systemic translatability of rodent neurotropic capsids in Old World primates are limited.…”

mentioning

confidence: 99%

“…In recent years, the field has focused on engineering novel capsids to address this problem, while simultaneously aiming to expand the therapeutic opportunity landscape for gene therapy into disorders not previously approachable with natural AAV serotypes. In parallel, the neuroscience community has also utilized novel AAVs, as several engineered AAV variants can traverse the restrictive blood-brainbarrier (BBB) to systemically deliver genetically-encoded tools to the rodent brain [14][15][16][17] , such as GCaMP to detect intracellular calcium gradients 18 . However, the majority of AAV capsid engineering efforts targeting the brain have thus far focused on increasing gene-transfer to the rodent central nervous system (CNS), and direct efforts in non-human primates (NHPs) are sparse.…”

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confidence: 99%

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Intravenous gene transfer throughout the brain of infant Old World primates using AAV

Chuapoco¹,

Flytzanis²,

Goeden³

et al. 2022

Preprint

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Adeno-associated viruses (AAVs) can enable robust and safe gene delivery to the mammalian central nervous system (CNS). While the scientific community has developed numerous neurotropic AAV variants for systemic gene-transfer to the rodent brain, there are few AAVs that efficiently access the CNS of higher order primates. We describe here AAV.CAP-Mac, an engineered AAV variant that enables systemic, brain-wide gene delivery in infants of two Old World primate species--the rhesus macaque (Macaca mulatta) and the green monkey (Chlorocebus sabaeus). We identified CAP-Mac using a multi-species selection strategy, initially screening our library in the adult common marmoset (Callithrix jacchus) and narrowing our pool of test-variants for another round of selection in infant macaques. In individual characterization, CAP-Mac robustly transduces human neurons in vitro and Old World primate neurons in vivo, where it targets all lobes of cortex, the cerebellum, and multiple subcortical regions of disease relevance. We use CAP-Mac for Brainbow-like multicolor labeling of macaque neurons throughout the brain, enabling morphological reconstruction of both medium spiny neurons and cortical pyramidal cells. Because of its broad distribution throughout the brain and high neuronal efficiency in infant Old World primates compared to AAV9, CAP-Mac shows promise for researchers and clinicians alike to unlock novel, noninvasive access to the brain for efficient gene transfer.

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Section: Cap-mac Is Enriched In the Infant Rhesus Macaque Cns Compared To Other Engineered Variantsmentioning

confidence: 69%

Section: Discussionmentioning

confidence: 99%

Section: Cap-mac Is Enriched In the Infant Rhesus Macaque Cns Compared To Other Engineered Variantsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Intravenous gene transfer throughout the brain of infant Old World primates using AAV

Chuapoco¹,

Flytzanis²,

Goeden³

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

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Bio–Nano Toolbox for Precision Alzheimer's Disease Gene Therapy

Liu,

Xia,

Zheng

et al. 2024

Advanced Materials

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Alzheimer's disease (AD) is the most burdensome aging‐associated neurodegenerative disorder, and its treatment encounters numerous failures during drug development. Although there are newly approved in‐market β‐amyloid targeting antibody solutions, pathological heterogeneity among patient populations still challenges the treatment outcome. Emerging advances in gene therapies offer opportunities for more precise personalized medicine; while, major obstacles including the pathological heterogeneity among patient populations, the puzzled mechanism for druggable target development, and the precision delivery of functional therapeutic elements across the blood–brain barrier remain and limit the use of gene therapy for central neuronal diseases. Aiming for “precision delivery” challenges, nanomedicine provides versatile platforms that may overcome the targeted delivery challenges for AD gene therapy. In this perspective, to picture a toolbox for AD gene therapy strategy development, the most recent advances from benchtop to clinics are highlighted, possibly available gene therapy targets, tools, and delivery platforms are outlined, their challenges as well as rational design elements are addressed, and perspectives in this promising research field are discussed.

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Precise Gene Knock‐In Tools with Minimized Risk of DSBs: A Trend for Gene Manipulation

Liu,

Kong,

Liu

et al. 2024

Advanced Science

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Gene knock‐in refers to the insertion of exogenous functional genes into a target genome to achieve continuous expression. Currently, most knock‐in tools are based on site‐directed nucleases, which can induce double‐strand breaks (DSBs) at the target, following which the designed donors carrying functional genes can be inserted via the endogenous gene repair pathway. The size of donor genes is limited by the characteristics of gene repair, and the DSBs induce risks like genotoxicity. New generation tools, such as prime editing, transposase, and integrase, can insert larger gene fragments while minimizing or eliminating the risk of DSBs, opening new avenues in the development of animal models and gene therapy. However, the elimination of off‐target events and the production of delivery carriers with precise requirements remain challenging, restricting the application of the current knock‐in treatments to mainly in vitro settings. Here, a comprehensive review of the knock‐in tools that do not/minimally rely on DSBs and use other mechanisms is provided. Moreover, the challenges and recent advances of in vivo knock‐in treatments in terms of the therapeutic process is discussed. Collectively, the new generation of DSBs‐minimizing and large‐fragment knock‐in tools has revolutionized the field of gene editing, from basic research to clinical treatment.

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AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset

Cited by 210 publications

References 61 publications

Intravenous gene transfer throughout the brain of infant Old World primates using AAV

Intravenous gene transfer throughout the brain of infant Old World primates using AAV

Bio–Nano Toolbox for Precision Alzheimer's Disease Gene Therapy

Precise Gene Knock‐In Tools with Minimized Risk of DSBs: A Trend for Gene Manipulation

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