In this continuing Bioengineering Research Partnership project, we seek to design improved antibody drugs for blocking aberrant signaling by epidermal growth factor receptor (EGFR) in cancer. A team with expertise in protein engineering (Wittrup, MIT), structural biology (Kuriyan, UC Berkeley), cell engineering (Lauffenburger, MIT), proteomics (White, MIT), and medical imaging (Larson, MSKCC) will further develop and characterize a panel of anti-EGFR molecules isolated in the previous project period. Our central hypothesis is that antibodies binding to domain IV of EGFR (anti-IV) block signaling in a ligand-independent manner, enabling antagonism of a common truncation mutant (EGFRvlll) that is not antagonized by the current generation of anti-EGFR antibodies. Anti-IV antibodies and fluorescence-based biophysical techniques will be used to further study the transduction of a symmetric ligand-bound extracellular dimerization event to an asymmetric active kinase dimer in the cytoplasm, recently characterized as part of this project. The molecular allosteric mechanisms that result in kinase domain activation upon ligand binding will be delineated so as to understand better how to block this process. We will apply mass spectrometry to simultaneously quantify the phosphorylation kinetics of hundreds of specific tyrosines in the signaling pathways downstream of EGFR, in particular looking for proximal evidence for eradication of EGFRvlll's effects by anti-IV antibodies. Enhanced receptor downregulation with multivalent antibodies will be systematically studied and exploited to reduce receptor number. Effects of anti-IV antibodies on migration and proliferation will be examined in autocrine cell lines. The kinetics of antibody penetration into tumor spheroids will be studied by 2-photon microscopy. In mouse xenograft models, the pharmacokinetics and pharmacodynamics of these antibodies will be studied by PET. This project is a systematic effort by a team of engineers, scientists, and physicians to develop a new generation of anticancer drugs through improved mechanistic understanding and blocking of a common class of uncontrolled growth signals sent by receptors on the surface of cancer cells.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA096504-09
Application #
8015219
Study Section
Special Emphasis Panel (ZRG1-ONC-G (03))
Program Officer
Welch, Anthony R
Project Start
2002-09-13
Project End
2013-01-31
Budget Start
2011-02-01
Budget End
2012-01-31
Support Year
9
Fiscal Year
2011
Total Cost
$581,866
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
001425594
City
Cambridge
State
MA
Country
United States
Zip Code
02139
Reddy, Raven J; Gajadhar, Aaron S; Swenson, Eric J et al. (2016) Early signaling dynamics of the epidermal growth factor receptor. Proc Natl Acad Sci U S A 113:3114-9
Traxlmayr, Michael W; Kiefer, Jonathan D; Srinivas, Raja R et al. (2016) Strong Enrichment of Aromatic Residues in Binding Sites from a Charge-neutralized Hyperthermostable Sso7d Scaffold Library. J Biol Chem 291:22496-22508
Kravchenko-Balasha, Nataly; Johnson, Hannah; White, Forest M et al. (2016) A Thermodynamic-Based Interpretation of Protein Expression Heterogeneity in Different Glioblastoma Multiforme Tumors Identifies Tumor-Specific Unbalanced Processes. J Phys Chem B 120:5990-7
Miller, Miles A; Sullivan, Ryan J; Lauffenburger, Douglas A (2016) Molecular Pathways: Receptor Ectodomain Shedding in Treatment, Resistance, and Monitoring of Cancer. Clin Cancer Res :
Reddy, Raven J; Curran, Timothy G; Zhang, Yi et al. (2016) Measurement of Phosphorylated Peptides with Absolute Quantification. Methods Mol Biol 1410:281-92
Tape, Christopher J; Ling, Stephanie; Dimitriadi, Maria et al. (2016) Oncogenic KRAS Regulates Tumor Cell Signaling via Stromal Reciprocation. Cell 165:910-20
de Picciotto, Seymour; Dickson, Paige M; Traxlmayr, Michael W et al. (2016) Design Principles for SuCESsFul Biosensors: Specific Fluorophore/Analyte Binding and Minimization of Fluorophore/Scaffold Interactions. J Mol Biol 428:4228-4241
Miller, Miles A; Oudin, Madeleine J; Sullivan, Ryan J et al. (2016) Reduced Proteolytic Shedding of Receptor Tyrosine Kinases Is a Post-Translational Mechanism of Kinase Inhibitor Resistance. Cancer Discov 6:382-99
Curran, Timothy G; Zhang, Yi; Ma, Daniel J et al. (2015) MARQUIS: a multiplex method for absolute quantification of peptides and posttranslational modifications. Nat Commun 6:5924
Kovacs, Erika; Zorn, Julie Anne; Huang, Yongjian et al. (2015) A structural perspective on the regulation of the epidermal growth factor receptor. Annu Rev Biochem 84:739-64

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