Dataset Information

Dysregulations in the levels of lactate dehydrogenase and pyruvate kinase upon IL-17A and echinomycin treatments in human primary keratinocytes

ABSTRACT: IL-17A driven hyperproliferation is a hallmark of psoriatic keratinocytes. The pathway analysis of shotgun proteomics data from human primary keratinocytes (HPKs) treated with IL-17A highlighted the role of increased levels of lactate dehydrogenase (LDH-A) and pyruvate kinase (PKM). We sought to validate these findings using multiple reaction monitoring (MRM) for LDH-A (6 peptides) and PKM (8 peptides). Moreover, speculating the role of the transcription factor HIF1a in regulation of LDH-A and PKM levels, we also probed these proteins upon inhibiting HIF1a activity with echinomycin in presence of IL-17A.

ORGANISM(S): Homo Sapiens

SUBMITTER: Medha Gayathri J Pai

PROVIDER: PXD040678 | panorama | Thu Aug 01 00:00:00 BST 2024

REPOSITORIES: PanoramaPublic

ACCESS DATA

Similar Datasets

Project description:Coordinated migration of B cells within and between secondary lymphoid tissues is required for robust antibody responses to infection or vaccination. Secondary lymphoid tissues normally expose B cells to a low O2 (hypoxic) environment. Recently, we have shown that human B cell migration is modulated by an O2-dependent molecular switch, centrally controlled by the hypoxia-induced (transcription) factor-1a (HIF1A), which can be disrupted by the immunosuppressive calcineurin inhibitor, cyclosporine A (CyA). However, the mechanisms by which low O2 environments attenuate B cell migration remain poorly defined. Proteomics analysis has linked CXCR4 chemokine receptor signaling to cytoskeletal rearrangement. We now hypothesize that the pathways linking the O2 sensing molecular switch to chemokine receptor signaling and cytoskeletal rearrangement would likely contain phosphorylation events, which are typically missed in traditional transcriptomic and/or proteomic analyses. Hence, we have performed a comprehensive phosphoproteomics analysis of human B cells treated with CyA after engagement of the chemokine receptor CXCR4 with CXCL12. Statistical analysis of the separate and synergistic effects of CyA and CXCL12 revealed 116 proteins whose abundance is driven by a synergistic interaction between CyA and CXCL12. Among these 116 proteins, Lymphocyte Specific Protein 1 (LSP1) was most interesting due to its original identification as a protein modulating neutrophil migration. We used our previously described algorithm BONITA to reveal a critical role for LSP1 in cytoskeletal rearrangement. Validating these modeling results, we show experimentally that LSP1 levels in B cells increase with low O2 exposure, and CyA treatment results in decreased LSP1 protein levels. This correlates with the increased chemotactic activity observed after CyA treatment. Lastly, we directly link LSP1 levels to chemotactic capacity, as shRNA knock-down of LSP1 results in significantly increased B cell chemotaxis at low O2 levels. These results directly link the CyA to LSP1-dependent cytoskeletal regulation, demonstrating a previously unrecognized mechanism by which CyA modulates human B cell migration.

Dataset Information

Dysregulations in the levels of lactate dehydrogenase and pyruvate kinase upon IL-17A and echinomycin treatments in human primary keratinocytes

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure