1. Biochemistry and Chemical Biology
  2. Cancer Biology
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Targeting RAS driven human cancer cells with antibodies to upregulated and essential cell-surface proteins

  1. Alexander John Martinko
  2. Charles Truillet
  3. Olivier Julien
  4. Juan Diaz
  5. Max A Horlbeck
  6. Gordon Whiteley
  7. Josip Blonder
  8. Jonathan S Weissman
  9. Sourav Bandyopadhyay
  10. Michael Evans
  11. James A Wells  Is a corresponding author
  1. University of California, San Francisco, United States
  2. Frederick National Laboratory for Cancer Research, United States
Research Article
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Cite as: eLife 2018;7:e31098 doi: 10.7554/eLife.31098

Abstract

While there have been tremendous efforts to target oncogenic RAS signaling from inside the cell, little effort has focused on the cell-surface. Here, we used quantitative surface proteomics to reveal a signature of proteins that are upregulated on cells transformed with KRASG12V, and driven by MAPK pathway signaling. We next generated a toolkit of recombinant antibodies to seven of these RAS induced proteins. We found that five of these proteins are broadly distributed on cancer cell lines harboring RAS mutations. In parallel, a cell-surface CRISPRi screen identified integrin and Wnt signaling proteins as critical to RAS transformed cells. We show that antibodies targeting CDCP1, a protein common to our proteomics and CRISPRi datasets, can be leveraged to deliver cytotoxic and immunotherapeutic payloads to RAS transformed cancer cells and report for RAS signaling status in vivo. Taken together, this work presents a technological platform for attacking RAS from outside the cell.

Article and author information

Author details

  1. Alexander John Martinko

    Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
    ORCID icon 0000-0002-4025-959X
  2. Charles Truillet

    Department of Radiology and Biomedical Imaging, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
  3. Olivier Julien

    Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
    ORCID icon 0000-0001-7068-7299
  4. Juan Diaz

    Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
  5. Max A Horlbeck

    Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
    ORCID icon 0000-0002-3875-871X
  6. Gordon Whiteley

    Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, United States
    Competing interests
    No competing interests declared.
  7. Josip Blonder

    Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, United States
    Competing interests
    No competing interests declared.
  8. Jonathan S Weissman

    Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
    ORCID icon 0000-0003-2445-670X
  9. Sourav Bandyopadhyay

    Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
    ORCID icon 0000-0003-3120-923X
  10. Michael Evans

    Department of Radiology and Biomedical Imaging, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
  11. James A Wells

    Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States
    For correspondence
    jim.wells@ucsf.edu
    Competing interests
    James A Wells, Celgene Corp provided some of the funding for this work.
    ORCID icon 0000-0001-8267-5519

Funding

National Institutes of Health (CA191018)

  • Alexander John Martinko
  • Olivier Julien
  • James A Wells

Celgene (GM097316)

  • Alexander John Martinko
  • James A Wells

National Institutes of Health (GM097316)

  • Alexander John Martinko
  • Olivier Julien
  • James A Wells

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (AN110806-03C) of the University of California San Francisco. Animals that were studied with PET/CT were anesthitized with a mixture of isoflurane/oxygen and humanely euthanized with CO2(g).

Reviewing Editor

  1. Jeffrey Settleman, Reviewing Editor, Calico Life Sciences, United States

Publication history

  1. Received: August 9, 2017
  2. Accepted: January 4, 2018
  3. Accepted Manuscript published: January 23, 2018 (version 1)
  4. Version of Record published: February 2, 2018 (version 2)

Copyright

© 2018, Martinko et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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