PXD040035

PXD040035 is an original dataset announced via ProteomeXchange.

Title	Casein kinase 2 activity is a host restriction factor for AAV transduction
Description	Despite rapid progress in the development of new therapies based on adeno-associated viral vectors (AAVs) in recent years, successful transduction of the human heart remains challenging. So far, the mechanisms that impede AAV transduction, especially in humans are poorly understood, hampering the introduction of new, effective gene therapy strategies. Therefore, the aim of this study was to identify and overcome the main cellular barriers to successful transduction in the heart, using iPSC-derived cardiomyocytes (iPSC-CMs), cardiac fibroblasts (iPSC-CFs), and primary endothelial cells (HAECs) to model vector-host interactions. Through phosphoproteome analysis of AAV9-transduced iPSC-CFs we established that casein kinase 2 (CK2) signalling is one of the most significantly affected pathways upon AAV exposure. Importantly, transient inhibition of CK2 activity with silmitasertib substantially enhanced the transduction rate of AAV2, AAV6 and AAV9 in all tested cell types (iPSC-CMs, iPSC-CFs and HAECs), demonstrating the versatility of this approach. CK2 inhibition improved the trafficking of AAVs through the cytoplasm and impaired DNA-damage response through destabilisation of Mre11, sensor of dsDNA breaks, that directly binds the ITRs of the vector genome. Silmitasertib treatment also allowed for improvement of transgene expression in already transduced iPSC-CFs, which retain AAV genomes in a functional, but probably silent form. Furthermore, our analysis revealed that AAV transduction may interfere with RNA processing pathways, identifying matrin-3 as a necessary factor for efficient transgene delivery. In summary, presented study provides new insights into the current understanding of the host-AAV vector interaction, identifying CK2 activity as a key barrier to efficient transduction and transgene expression, therefore offering a promising strategy to improve the outcome of AAV-based therapies in the future.
HostingRepository	PRIDE
AnnounceDate	2024-03-28
AnnouncementXML	Submission_2024-03-28_06:50:03.938.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Luisa Marie Schmidt
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2023-02-10 06:23:44	ID requested
⏵ 1	2024-03-28 06:50:04	announced

10.1016/J.YMTHE.2023.11.010;

submitter keyword: phosphopeptides, cardiomyocytes, CK2,iPSC, AAV transduction

Agnieszka Jaźwa-Kusior
contact affiliation	1Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland
contact email	agnieszka.jazwa@uj.edu.pl
lab head
Luisa Marie Schmidt
contact affiliation	CECAD research center, Institue of Genetics, University of Cologne
contact email	luisa.schmidt@uni-koeln.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2024/03/PXD040035

PRIDE project URI

• PRIDE
  1. PXD040035
     1. Label: PRIDE project
     2. Name: Casein kinase 2 activity is a host restriction factor for AAV transduction