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LipidFinder: A computational workflow for discovery of lipids identifies eicosanoid-phosphoinositides in platelets
Anne O’Connor, … , Stuart M. Allen, Valerie B. O’Donnell
Anne O’Connor, … , Stuart M. Allen, Valerie B. O’Donnell
Published April 6, 2017
Citation Information: JCI Insight. 2017;2(7):e91634. https://doi.org/10.1172/jci.insight.91634.
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Resource and Technical Advance Inflammation Metabolism

LipidFinder: A computational workflow for discovery of lipids identifies eicosanoid-phosphoinositides in platelets

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Abstract

Accurate and high-quality curation of lipidomic datasets generated from plasma, cells, or tissues is becoming essential for cell biology investigations and biomarker discovery for personalized medicine. However, a major challenge lies in removing artifacts otherwise mistakenly interpreted as real lipids from large mass spectrometry files (>60 K features), while retaining genuine ions in the dataset. This requires powerful informatics tools; however, available workflows have not been tailored specifically for lipidomics, particularly discovery research. We designed LipidFinder, an open-source Python workflow. An algorithm is included that optimizes analysis based on users’ own data, and outputs are screened against online databases and categorized into LIPID MAPS classes. LipidFinder outperformed three widely used metabolomics packages using data from human platelets. We show a family of three 12-hydroxyeicosatetraenoic acid phosphoinositides (16:0/, 18:1/, 18:0/12-HETE-PI) generated by thrombin-activated platelets, indicating crosstalk between eicosanoid and phosphoinositide pathways in human cells. The software is available on GitHub (https://github.com/cjbrasher/LipidFinder), with full userguides.

Authors

Anne O’Connor, Christopher J. Brasher, David A. Slatter, Sven W. Meckelmann, Jade I. Hawksworth, Stuart M. Allen, Valerie B. O’Donnell

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Figure 6

MS identification of eicosanoid-phosphoinositide lipids generated by platelets.

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MS identification of eicosanoid-phosphoinositide lipids generated by pla...
(A) Orbitrap MS of HETE-PIs in platelet lipid extracts at 60,000 resolution (at m/z 400) showing elution of ions with m/z values corresponding to 18:0, 18:1, and 16:0/12-HETE-PIs. (B) MS of the putative HETE-PI ions. Single ions are shown at expected m/z values, circled, for the corresponding lipids in A. (C) MS/MS of the two most abundant HETE-PIs. Ions from 12-HETE (m/z 179, 319), PI headgroup (153,241, 315), and sn1 fatty acids (283 and 281 for 18:0 and 18:1, respectively) are seen. (D and E) LC/MS/MS separation of thrombin-activated (D) or basal (E) platelet lipid extracts monitoring MRM transitions corresponding to HETE-PEs. HETE-PIs are only detected following activation of platelets.

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