PMID- 33441336
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20230228
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 95
IP  - 7
DP  - 2021 Mar 10
TI  - Adeno-associated Virus (AAV) Capsid Chimeras with Enhanced Infectivity Reveal a 
      Core Element in the AAV Genome Critical for both Cell Transduction and Capsid 
      Assembly.
LID - JVI.02023-20 [pii]
LID - 10.1128/JVI.02023-20 [doi]
LID - e02023-20
AB  - Adeno-associated viruses (AAV) have attracted significant attention in the field 
      of gene and cell therapy due to highly effective delivery of therapeutic genes 
      into human cells. The ability to generate recombinant AAV vectors compromised of 
      unique or substituted protein sequences has led to the development of capsid 
      variants with improved therapeutic properties. Seeking novel AAV vectors capable 
      of enhanced transduction for therapeutic applications, we have developed a series 
      of unique capsid variants termed AAV X-Vivo (AAV-XV) derived from chimeras of 
      AAV12 VP1/2 sequences and the VP3 sequence of AAV6. These AAV variants showed 
      enhanced infection of human primary T cells, hematopoietic stem cells, and 
      neuronal cell lines over wildtype parental viruses, and superiority over AAV6 for 
      genomic integration of DNA sequences by AAV alone or in combination with CRISPR 
      gene editing. AAV-XV variants demonstrate transduction efficiency equivalent to 
      AAV6 at multiplicities of infection 2 logs lower, enabling T cell engineering at 
      low AAV doses. The protein coding sequence of these novel AAV chimeras revealed 
      disruptions within the assembly-activating protein (AAP) which likely accounted 
      for observed lower virus yield. A series of genome alterations, reverting the AAP 
      sequence back to wildtype AAV6, had a negative impact on the enhanced 
      transduction seen with AAV-VX, indicating overlapping functions within this 
      sequence for both viral assembly and effective T cell transduction. Our findings 
      show these AAV-XV variants are highly efficient at cell transduction at low dose 
      and demonstrates the importance of the AAP coding region in both viral particle 
      assembly and cell infection.IMPORTANCE A major hurdle to the therapeutic 
      potential of AAV in gene therapy lies in achieving clinically meaningful AAV 
      doses, and secondarily, ability to manufacture commercially viable titers of AAV 
      to support this. By virtue of neutralizing antibodies against AAV that impede 
      patient repeat-dosing, the dose of AAV for in vivo gene delivery has been high, 
      which has resulted in unfortunate recent safety concerns and deaths in patients 
      given higher-dose AAV gene therapy. We have generated new AAV variants possessing 
      unique combinations of capsid proteins for gene and cell therapy applications 
      termed AAV-XV, which have high levels of cell transduction and gene delivery at 
      lower MOI. Furthermore, we demonstrate a novel finding, and an important 
      consideration for recombinant AAV design, that a region of the AAV genome 
      encoding the capsid viral protein and AAP is critical for both virus yield and 
      the enhancement of infection/transduction.
CI  - Copyright (c) 2021 American Society for Microbiology.
FAU - Viney, Lydia
AU  - Viney L
AD  - Intima Bioscience, New York, New York, USA.
FAU - Burckstummer, Tilmann
AU  - Burckstummer T
AD  - Intima Bioscience, New York, New York, USA.
FAU - Eddington, Courtnee
AU  - Eddington C
AD  - Department of Biochemistry and Molecular Biology, Center for Structural Biology, 
      McKnight Brain Institute, University of Florida, Gainesville, Florida, USA.
FAU - Mietzsch, Mario
AU  - Mietzsch M
AD  - Department of Biochemistry and Molecular Biology, Center for Structural Biology, 
      McKnight Brain Institute, University of Florida, Gainesville, Florida, USA.
FAU - Choudhry, Modassir
AU  - Choudhry M
AD  - Intima Bioscience, New York, New York, USA.
FAU - Henley, Tom
AU  - Henley T
AD  - Intima Bioscience, New York, New York, USA tom@intimabioscience.com 
      mckenna@ufl.edu.
FAU - Agbandje-McKenna, Mavis
AU  - Agbandje-McKenna M
AD  - Department of Biochemistry and Molecular Biology, Center for Structural Biology, 
      McKnight Brain Institute, University of Florida, Gainesville, Florida, USA 
      tom@intimabioscience.com mckenna@ufl.edu.
LA  - eng
PT  - Journal Article
DEP - 20210113
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
SB  - IM
PMC - PMC8092683
EDAT- 2021/01/15 06:00
MHDA- 2021/01/15 06:01
PMCR- 2021/03/10
CRDT- 2021/01/14 05:33
PHST- 2021/01/14 05:33 [entrez]
PHST- 2021/01/15 06:00 [pubmed]
PHST- 2021/01/15 06:01 [medline]
PHST- 2021/03/10 00:00 [pmc-release]
AID - JVI.02023-20 [pii]
AID - 02023-20 [pii]
AID - 10.1128/JVI.02023-20 [doi]
PST - ppublish
SO  - J Virol. 2021 Mar 10;95(7):e02023-20. doi: 10.1128/JVI.02023-20. Epub 2021 Jan 
      13.