PXD051824-1

PXD051824 is an original dataset announced via ProteomeXchange.

Title	Cytotoxic mechanisms of pemetrexed and HDAC inhibition in non-small cell lung cancer cells
Description	The cytotoxic effects of thymidylate synthase (TYMS) inhibitors, such as the multitarget antifolate pemetrexed (PMX), are not fully understood. Recent studies suggest that the combination of PMX with histone deacetylase inhibitors may be effective in the treatment of non- small cell lung cancer (NSCLC). To investigate this further, A549 NSCLC cells were exposed to various combinations of PMX and the HDAC inhibitor MS275 (entinostat) and Vorinostat (SAHA) and monitored for cell survival, cell cycle alterations and genotoxic markers. MS275 increased PMX sensitivity in A549 cells when added after preincubation with PMX. Both HeLa (p53 negative) and A549 (p53-naïve) exhibited robust activation of γH2AX upon treatment with the combination. Interestingly, CRISPR-Cas9 UNG knockout did not affect γH2AX activation or sensitivity to PMX. Proteomic analysis revealed that MS275 altered expression of known PMX targets, several proteins involved in pyrimidine metabolism, and several DNA repair proteins (RNaseH2, TOP1, RAD51C, PCNA). Contra to the prevailing model of antifolate toxicity mediated by uracil excision and repeated futile repair cycles caused by reincorporation of uracil, we propose that ribonucleotide incorporation in nuclear and mitochondrial DNA is a major contributor to cytotoxicity antifolates such as PMX, and likely also for fluorinated pyrimidine analogs. The DNA lesions induced by these agents and how their processing can be modulated by HDAC inhibition to potentiate cytotoxicity, warrants further investigation in clinical models.
HostingRepository	PRIDE
AnnounceDate	2024-12-19
AnnouncementXML	Submission_2024-12-18_16:12:49.565.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Animesh Sharma
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue
Instrument	timsTOF Pro

Revision	Datetime	Status	ChangeLog Entry
0	2024-04-28 09:14:20	ID requested
⏵ 1	2024-12-18 16:12:49	announced
2	2025-01-20 04:15:01	announced	2025-01-20: Updated project metadata.

Dataset with its publication pending

submitter keyword: BER, double strand breaks, base excision repair, histone deacetylase,AP, dihydrofolate reductase, dUTPase DUT, DHFR, apyrimidinic, deoxyuridine triphosphate nucleotide hydrolase, GART, trifunctional purine biosynthetic protein adenosine 3, HDAC, HDACi, DSB

Tobias Iveland
contact affiliation	Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, N-7006 Trondheim, Norway
contact email	tobias.iveland@gmail.com
lab head
Animesh Sharma
contact affiliation	Engineer at NTNU, Norway
contact email	sharma.animesh@gmail.com
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/2024/12/PXD051824

PRIDE project URI

• PRIDE
  1. PXD051824
     1. Label: PRIDE project
     2. Name: Cytotoxic mechanisms of pemetrexed and HDAC inhibition in non-small cell lung cancer cells