Scratching the surface of RGD-directed AAV capsid engineering

Zolotukhin, Sergei; Trivedi, Prasad D.; Corti, Manuela; Byrne, Barry J.

doi:10.1016/j.ymthe.2021.10.020

Cited by 5 publications

(3 citation statements)

References 9 publications

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“…In line with this idea, a recent paper described the superiority of RGD motif-containing AAVs for the transduction of myocytes in mice and non-human primates 51 , concurrent with data reported before by some of us for the RGD-displaying AAV9P1 variant in the musculature of mice 52 . However, the relation between integrin (subtype) expression and AAV biology is likely to be very complex 53,54 . For instance, while our experiments revealed a prominent decrease of fluorescent intensity when iNSCs transduced with AAV9P1-YFP were treated with higher concentrations of a selective integrin αvβ8 ligand, the overall fraction of transduced cells appeared unchanged.…”

Section: Discussionmentioning

confidence: 99%

Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening

Brüstle

Börner

Stüllein

et al. 2022

Sci Rep

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Human brain cells generated by in vitro cell programming provide exciting prospects for disease modeling, drug discovery and cell therapy. These applications frequently require efficient and clinically compliant tools for genetic modification of the cells. Recombinant adeno-associated viruses (AAVs) fulfill these prerequisites for a number of reasons, including the availability of a myriad of AAV capsid variants with distinct cell type specificity (also called tropism). Here, we harnessed a customizable parallel screening approach to assess a panel of natural or synthetic AAV capsid variants for their efficacy in lineage-related human neural cell types. We identified common lead candidates suited for the transduction of directly converted, early-stage induced neural stem cells (iNSCs), induced pluripotent stem cell (iPSC)-derived later-stage, radial glia-like neural progenitors, as well as differentiated astrocytic and mixed neuroglial cultures. We then selected a subset of these candidates for functional validation in iNSCs and iPSC-derived astrocytes, using shRNA-induced downregulation of the citrate transporter SLC25A1 and overexpression of the transcription factor NGN2 for proofs-of-concept. Our study provides a comparative overview of the susceptibility of different human cell programming-derived brain cell types to AAV transduction and a critical discussion of the assets and limitations of this specific AAV capsid screening approach.

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Section: Discussionmentioning

confidence: 99%

Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening

Brüstle

Börner

Stüllein

et al. 2022

Sci Rep

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“…The following approaches might be helpful in making AdV vectors more efficient. Modifying RGD-directed structural AdV might be an alternative combinatorial peptide display which would lead to higher specificity and efficient endocytosis, but it might have to assess the safety profile of this approach related to the immune response as well, like discussed previously for rAAV related optimizations [200]. A more promising approach involves integrating the promoter regions of genes highly active in cancer cells, such as human telomerase reverse transcriptase and cyclooxygenase 2, into locations that govern adenovirus replication gene expression [201,202].…”

Section: Recent Progress In the Fieldmentioning

confidence: 99%

Evolving Horizons: Adenovirus Vectors’ Timeless Influence on Cancer, Gene Therapy and Vaccines

Trivedi,

Byrne,

Corti

2023

Viruses

Self Cite

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Efficient and targeted delivery of a DNA payload is vital for developing safe gene therapy. Owing to the recent success of commercial oncolytic vector and multiple COVID-19 vaccines, adenovirus vectors are back in the spotlight. Adenovirus vectors can be used in gene therapy by altering the wild-type virus and making it replication-defective; specific viral genes can be removed and replaced with a segment that holds a therapeutic gene, and this vector can be used as delivery vehicle for tissue specific gene delivery. Modified conditionally replicative–oncolytic adenoviruses target tumors exclusively and have been studied in clinical trials extensively. This comprehensive review seeks to offer a summary of adenovirus vectors, exploring their characteristics, genetic enhancements, and diverse applications in clinical and preclinical settings. A significant emphasis is placed on their crucial role in advancing cancer therapy and the latest breakthroughs in vaccine clinical trials for various diseases. Additionally, we tackle current challenges and future avenues for optimizing adenovirus vectors, promising to open new frontiers in the fields of cell and gene therapies.

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“…Importantly, the myotropic AAVs [15][16][17] identi ed by muscle-directed high-throughput screening (HTS) were shown to share an Arg-Gly-Asp (RGD) motif, presumably targeting the integrin complex [17][18][19][20] . Integrins are a group of heterodimeric proteins composed of an α-and a β subunit that serve various cellular functions, including cell adhesion, cell migration, and cell signaling 21 .…”

Section: Introductionmentioning

confidence: 99%

An integrin-targeting AAV developed using a novel computational rational design methodology presents improved targeting of the skeletal muscle and reduced liver tropism

Hong,

Suel,

Poupiot

et al. 2023

Preprint

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Current adeno-associated virus (AAV) gene therapy using nature-derived AAVs is limited by non-optimal tissue targeting. In the treatment of muscular diseases (MD), high doses are therefore often required, but can lead to severe adverse effects. To lower treatment doses, we rationally designed an AAV that specifically targets skeletal muscle. We employed a novel computational design that integrated binding motifs of integrin alpha V beta 6 (αVβ6) into a liver-detargeting AAV capsid backbone to target the human αVβ6 complex – a selected AAV receptor for skeletal muscle. After sampling the low-energy capsid mutants, all in silico designed AAVs showed higher productivity compared to their parent. We confirmed in vitro that the enhanced transduction is due to the binding to the αVβ6 complex. Thanks to inclusion of αVβ6-binding motifs, the designed AAVs exhibited enhanced transduction efficacy in human differentiated myotubes as well as in murine skeletal muscles in vivo. One notable variant, LICA1, showed similar muscle transduction to other published myotropic AAVs, while being significantly more strongly liver-detargeted. We further examined the efficacy of LICA1, in comparison to AAV9, in delivering therapeutic transgenes in two mouse MD models at a low dose of 5E12 vg/kg. At this dose, AAV9 was suboptimal, while LICA1 transduced effectively and significantly better than AAV9 in all tested muscles. Consequently, LICA1 corrected the myopathology, restored global transcriptomic dysregulation, and improved muscle functionality. These results underline the potential of our design method for AAV engineering and demonstrate the relevance of the novel AAV variant for gene therapy treatment of MD.

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Scratching the surface of RGD-directed AAV capsid engineering

Cited by 5 publications

References 9 publications

Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening

Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening

Evolving Horizons: Adenovirus Vectors’ Timeless Influence on Cancer, Gene Therapy and Vaccines

An integrin-targeting AAV developed using a novel computational rational design methodology presents improved targeting of the skeletal muscle and reduced liver tropism

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