PXD003519-1

PXD003519 is an original dataset announced via ProteomeXchange.

Title	Single Cell Immuno-laser Microdissection Coupled to Label-free Proteomics to Reveal the Proteotypes of Human Brain Cells After Ischemia
Description	Cerebral ischemia entails rapid tissue damage in the affected brain area causing devastating neurological disorders. The use of biomarkers in stroke might have a major impact in patients’ management, both in the diagnosis and the prognosis. Thus, the analysis of the brain proteome is a straightforward approach for the identification of specific biomarkers and for the detection of potential new therapeutic targets for neuroprotection or recovery strategies. In this study, post-mortem brain slices from ischemic stroke patients were obtained corresponding to infarcted (IC) and contralateral (CL) areas. By means of laser microdissection, neurons and blood brain barrier structures (BBB) were isolated and analyzed in a label-free manner on a LTQ Orbitrap Velos Pro MS following a gas phase fractionation approach for precursor ion selection. Ninety one proteins were identified only in neurons, 261 proteins only in BBB and 260 proteins in both cell types (peptides≥2). Gene ontology analyses revealed the alteration of metabolic processes after brain ischemia and also changes regarding catalytic activity and binding. Antioxidant activity and enzyme regulator activity appeared as cell-specific altered molecular functions. A total of 25 proteins showing p<0.05 and fold-change>|2| between IC and CL areas were considered significant in this study: 11 in neurons, 11 in BBB and 3 in both cell types. Twelve of these proteins were selected as candidates and analyzed by immunohistochemistry in independent brains. The MS findings were verified for neuronal SAHH2 and SRSF1 and for EAA2 in both cell types. In addition, SAHH2 showed its potential as prognostic biomarker when analyzed early in plasma of ischemic stroke patients. Therefore, our results will contribute to increase the knowledge about the molecular mechanisms of ischemic stroke pathology and launch new proteins that might represent putative biomarkers of brain ischemia or therapeutic targets.
HostingRepository	PRIDE
AnnounceDate	2017-12-04
AnnouncementXML	Submission_2017-12-04_02:09:57.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD003519
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Víctor Llombart
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	Oxidation; Carbamidomethyl
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2016-01-25 03:49:16	ID requested
⏵ 1	2017-12-04 02:09:59	announced
2	2024-10-22 04:40:04	announced	2024-10-22: Updated project metadata.

Garc, í, a-Berrocoso T, Llombart V, Col, à, s-Camp, à, s L, Hainard A, Licker V, Penalba A, Ramiro L, Simats A, Bustamante A, Mart, í, nez-Saez E, Canals F, Sanchez JC, Montaner J, Single Cell Immuno-Laser Microdissection Coupled to Label-Free Proteomics to Reveal the Proteotypes of Human Brain Cells After Ischemia. Mol Cell Proteomics, 17(1):175-189(2018) [pubmed]

curator keyword: Biomedical

submitter keyword: stroke, brain ischemia, laser micro-dissection, label-free, proteomics, biomarker

Joan Montaner
contact affiliation	Neurovascular Research Laboratory, Vall Hebron Institute of Research, Barcelona, Spain.
contact email	montaner.villalonga@gmail.com
lab head
Víctor Llombart
contact affiliation	Vall Hebrón Institute of Research
contact email	llombs@hotmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD003519
     1. Label: PRIDE project
     2. Name: Single Cell Immuno-laser Microdissection Coupled to Label-free Proteomics to Reveal the Proteotypes of Human Brain Cells After Ischemia