Molecular Systems Biology (Sep 2021)
A highly multiplexed quantitative phosphosite assay for biology and preclinical studies
- Hasmik Keshishian,
- E Robert McDonald,
- Filip Mundt,
- Randy Melanson,
- Karsten Krug,
- Dale A Porter,
- Luke Wallace,
- Dominique Forestier,
- Bokang Rabasha,
- Sara E Marlow,
- Judit Jane‐Valbuena,
- Ellen Todres,
- Harrison Specht,
- Margaret Lea Robinson,
- Pierre M Jean Beltran,
- Ozgun Babur,
- Meagan E Olive,
- Javad Golji,
- Eric Kuhn,
- Michael Burgess,
- Melanie A MacMullan,
- Tomas Rejtar,
- Karen Wang,
- DR Mani,
- Shankha Satpathy,
- Michael A Gillette,
- William R Sellers,
- Steven A Carr
Affiliations
- Hasmik Keshishian
- Broad Institute of Massachusetts Institute of Technology and Harvard
- E Robert McDonald
- Novartis Institute of Biomedical Research
- Filip Mundt
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Randy Melanson
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Karsten Krug
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Dale A Porter
- Novartis Institute of Biomedical Research
- Luke Wallace
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Dominique Forestier
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Bokang Rabasha
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Sara E Marlow
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Judit Jane‐Valbuena
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Ellen Todres
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Harrison Specht
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Margaret Lea Robinson
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Pierre M Jean Beltran
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Ozgun Babur
- Computer Science Department, University of Massachusetts Boston
- Meagan E Olive
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Javad Golji
- Novartis Institute of Biomedical Research
- Eric Kuhn
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Michael Burgess
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Melanie A MacMullan
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Tomas Rejtar
- Novartis Institute of Biomedical Research
- Karen Wang
- Novartis Institute of Biomedical Research
- DR Mani
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Shankha Satpathy
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Michael A Gillette
- Broad Institute of Massachusetts Institute of Technology and Harvard
- William R Sellers
- Broad Institute of Massachusetts Institute of Technology and Harvard
- Steven A Carr
- Broad Institute of Massachusetts Institute of Technology and Harvard
- DOI
- https://doi.org/10.15252/msb.202010156
- Journal volume & issue
-
Vol. 17,
no. 9
pp. 1 – 22
Abstract
Abstract Reliable methods to quantify dynamic signaling changes across diverse pathways are needed to better understand the effects of disease and drug treatment in cells and tissues but are presently lacking. Here, we present SigPath, a targeted mass spectrometry (MS) assay that measures 284 phosphosites in 200 phosphoproteins of biological interest. SigPath probes a broad swath of signaling biology with high throughput and quantitative precision. We applied the assay to investigate changes in phospho‐signaling in drug‐treated cancer cell lines, breast cancer preclinical models, and human medulloblastoma tumors. In addition to validating previous findings, SigPath detected and quantified a large number of differentially regulated phosphosites newly associated with disease models and human tumors at baseline or with drug perturbation. Our results highlight the potential of SigPath to monitor phosphoproteomic signaling events and to nominate mechanistic hypotheses regarding oncogenesis, response, and resistance to therapy.
Keywords
