Inhibition of DNA-dependent protein kinase catalytic subunit boosts rAAV transduction of polarized human airway epithelium

Kang Ning; Xiujuan Zhang; Zehua Feng; Siyuan Hao; Cagla Aksu Kuz; Fang Cheng; Soo Yuen Park; Shane McFarlin; John F. Engelhardt; Ziying Yan; Jianming Qiu

doi:10.1016/j.omtm.2023.101115

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Inhibition of DNA-dependent protein kinase catalytic subunit boosts rAAV transduction of polarized human airway epithelium

Journal article

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Inhibition of DNA-dependent protein kinase catalytic subunit boosts rAAV transduction of polarized human airway epithelium

Kang Ning, Xiujuan Zhang, Zehua Feng, Siyuan Hao, Cagla Aksu Kuz, Fang Cheng, Soo Yuen Park, Shane McFarlin, John F. Engelhardt, Ziying Yan, …

Molecular therapy. Methods & clinical development, Vol.31, 101115

12/2023

DOI: 10.1016/j.omtm.2023.101115

PMCID: PMC10568418

PMID: 37841417

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Abstract

Adeno-associated virus (AAV)2.5T was selected from the directed evolution of AAV capsid library in human airway epithelia. This study found that recombinant (r)AAV2.5T transduction of well-differentiated primary human airway epithelia induced a DNA damage response (DDR), characterized by the phosphorylation of replication protein A32 (RPA32), histone variant H2AX (H2A histone family member X), and all three phosphatidylinositol 3-kinase-related kinases (PIKKs): Ataxia-telangiectasia mutated kinase (ATM), Ataxia telangiectasia and Rad3-related kinase (ATR), and DNA-dependent protein kinase catalytic subunit (DNA-PKcs). While suppressing the expression of ATR by a specific pharmacological inhibitor or targeted gene silencing inhibited rAAV2.5T transduction, DNA-PKcs inhibition or targeted gene silencing significantly increased rAAV2.5T transgene expression. Notably, DNA-PKcs inhibitors worked as a “booster” to further increase rAAV2.5T transgene expression after treatment with doxorubicin and did not compromise epithelial integrity. Thus, our study provides evidence that DDR is associated with rAAV transduction in well-differentiated human airway epithelia, and DNA-PKcs inhibition has the potential to boost rAAV transduction. These findings highlight that the application of DDR inhibition-associated pharmacological interventions has the potential to increase rAAV transduction and thus to reduce the required vector dose. [Display omitted]

Details

Title: Subtitle: Inhibition of DNA-dependent protein kinase catalytic subunit boosts rAAV transduction of polarized human airway epithelium
Creators: Kang Ning - University of Kansas Medical Center
Xiujuan Zhang - University of Kansas Medical Center
Zehua Feng - University of Iowa
Siyuan Hao - University of Kansas Medical Center
Cagla Aksu Kuz - University of Kansas Medical Center
Fang Cheng - University of Kansas Medical Center
Soo Yuen Park - Department of Anatomy and Cell Biology, University of Iowa, Iowa City, IA 52242, USA
Shane McFarlin - University of Kansas Medical Center
John F. Engelhardt - University of Iowa
Ziying Yan - University of Iowa
Jianming Qiu - University of Kansas Medical Center
Resource Type: Journal article
Publication Details: Molecular therapy. Methods & clinical development, Vol.31, 101115
DOI: 10.1016/j.omtm.2023.101115
PMID: 37841417
PMCID: PMC10568418
NLM abbreviation: Mol Ther Methods Clin Dev
ISSN: 2329-0501
eISSN: 2329-0501
Publisher: Elsevier Inc
Grant note: DOI: 10.13039/100000060, name: NIAID, award: AI150877, AI156448; DOI: 10.13039/100000062, name: NIDDK, award: DK054759; DOI: 10.13039/100008893, name: University of Iowa; DOI: 10.13039/100000002, name: National Institutes of Health, award: S10 OD 023625
Language: English
Electronic publication date: 09/2023
Date published: 12/2023
Academic Unit: Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
Record Identifier: 9984472656802771

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