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NIH Research Festival

September 16 – 18, 2015

Targeted proteomics-driven computational modeling of macrophage S1P chemosensing

Thursday, September 17, 2015 – Poster Session II
12:00 – 1:30 p.m.

FAES Terrace

NIAID

SYSBIO-4

Authors

  • NP Manes
  • BR Angermann
  • M Koppenol-Raab
  • E An
  • VH Sjoelund
  • J Sun
  • M Ishii
  • RN Germain
  • M Meier-Schellersheim
  • A Nita-Lazar

Abstract

Osteoclasts are monocyte-derived multinuclear cells that directly attach to and resorb bone. Sphingosine-1-phosphate (S1P) regulates bone resorption by functioning as both a chemoattractant and chemorepellent of osteoclast precursors through two G-protein coupled receptors that antagonize each other in an S1P-concentration dependent manner. To quantitatively explore the behavior of this chemosensing pathway, we applied targeted proteomics, transcriptomics, and rule-based pathway modeling to the S1P pathway using the Simmune toolset. RAW264.7 cells (a mouse monocyte/macrophage cell line) were used as model osteoclast precursors, RNA-seq was used to identify expressed target proteins, and selected reaction monitoring (SRM) mass spectrometry using internal peptide standards was used to perform absolute abundance measurements of pathway proteins. The resulting transcript and protein abundance values were strongly correlated. Measured protein abundance values, used as simulation input parameters, led to in silico pathway behavior matching in vitro measurements. Moreover, once model parameters were established, even simulated responses towards stimuli that were not used for parameterization were consistent with experimental findings. These findings demonstrate the feasibility and value of combining targeted mass spectrometry with pathway modeling for advancing biological insight.

Scientific Focus Area: Systems Biology

This page was last updated on Friday, March 26, 2021

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