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Thomas C Harding,
Li Long,
Servando Palencia,
Hongbing Zhang,
Ali Sadra,
Kevin Hestir,
Namrata Patil,
Anita Levin,
Amy W Hsu,
Deborah Charych,
Thomas Brennan,
James Zanghi,
Robert Halenbeck,
Shannon A Marshall,
Minmin Qin, Stephen K Doberstein,
Diane Hollenbaugh,
W Michael Kavanaugh,
Lewis T Williams,
Kevin P Baker
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ABSTRACT: The fibroblast growth factor (FGF) pathway promotes tumor growth and angiogenesis in many solid tumors. Although there has long been interest in FGF pathway inhibitors, development has been complicated: An effective FGF inhibitor must block the activity of multiple mitogenic FGF ligands but must spare the metabolic hormone FGFs (FGF-19, FGF-21, and FGF-23) to avoid unacceptable toxicity. To achieve these design requirements, we engineered a soluble FGF receptor 1 Fc fusion protein, FP-1039. FP-1039 binds tightly to all of the mitogenic FGF ligands, inhibits FGF-stimulated cell proliferation in vitro, blocks FGF- and vascular endothelial growth factor (VEGF)-induced angiogenesis in vivo, and inhibits in vivo growth of a broad range of tumor types. FP-1039 antitumor response is positively correlated with RNA levels of FGF2, FGF18, FGFR1c, FGFR3c, and ETV4; models with genetic aberrations in the FGF pathway, including FGFR1-amplified lung cancer and FGFR2-mutated endometrial cancer, are particularly sensitive to FP-1039-mediated tumor inhibition. FP-1039 does not appreciably bind the hormonal FGFs, because these ligands require a cell surface co-receptor, klotho or β-klotho, for high-affinity binding and signaling. Serum calcium and phosphate levels, which are regulated by FGF-23, are not altered by administration of FP-1039. By selectively blocking nonhormonal FGFs, FP-1039 treatment confers antitumor efficacy without the toxicities associated with other FGF pathway inhibitors.
Science translational medicine 03/2013; 5(178):178ra39. · 7.80 Impact Factor
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Haishan Lin,
Ernestine Lee,
Kevin Hestir,
Cindy Leo,
Minmei Huang,
Elizabeth Bosch,
Robert Halenbeck,
Ge Wu,
Aileen Zhou,
Dirk Behrens,
Diane Hollenbaugh,
Thomas Linnemann,
Minmin Qin,
Justin Wong,
Keting Chu, Stephen K Doberstein,
Lewis T Williams
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ABSTRACT: To understand the system of secreted proteins and receptors involved in cell-cell signaling, we produced a comprehensive set of recombinant secreted proteins and the extracellular domains of transmembrane proteins, which constitute most of the protein components of the extracellular space. Each protein was tested in a suite of assays that measured metabolic, growth, or transcriptional responses in diverse cell types. The pattern of responses across assays was analyzed for the degree of functional selectivity of each protein. One of the highly selective proteins was a previously undescribed ligand, designated interleukin-34 (IL-34), which stimulates monocyte viability but does not affect responses in a wide spectrum of other assays. In a separate functional screen, we used a collection of extracellular domains of transmembrane proteins to discover the receptor for IL-34, which was a known cytokine receptor, colony-stimulating factor 1 (also called macrophage colony-stimulating factor) receptor. This systematic approach is thus useful for discovering new ligands and receptors and assessing the functional selectivity of extracellular regulatory proteins.
Science 06/2008; 320(5877):807-11. · 31.20 Impact Factor
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Jonathan Zalevsky,
Thomas Secher,
Sergei A Ezhevsky,
Laure Janot,
Paul M Steed,
Christopher O'Brien,
Araz Eivazi,
James Kung,
Duc-Hanh T Nguyen, Stephen K Doberstein,
François Erard,
Bernhard Ryffel,
David E Szymkowski
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ABSTRACT: TNF is a pleiotropic cytokine required for normal development and function of the immune system; however, TNF overexpression also induces inflammation and is associated with autoimmune diseases. TNF exists as both a soluble and a transmembrane protein. Genetic studies in mice have suggested that inflammation in disease models involves soluble TNF (solTNF) and that maintenance of innate immune function involves transmembrane TNF (tmTNF). These findings imply that selective pharmacologic inhibition of solTNF may be anti-inflammatory and yet preserve innate immunity to infection. To address this hypothesis, we now describe dominant-negative inhibitors of TNF (DN-TNFs) as a new class of biologics that selectively inhibits solTNF. DN-TNFs blocked solTNF activity in human and mouse cells, a human blood cytokine release assay, and two mouse arthritis models. In contrast, DN-TNFs neither inhibited the activity of human or mouse tmTNF nor suppressed innate immunity to Listeria infection in mice. These results establish DN-TNFs as the first selective inhibitors of solTNF, demonstrate that inflammation in mouse arthritis models is primarily driven by solTNF, and suggest that the maintenance of tmTNF activity may improve the therapeutic index of future anti-inflammatory agents.
The Journal of Immunology 09/2007; 179(3):1872-83. · 5.79 Impact Factor
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Ge Wu, Stephen K Doberstein
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ABSTRACT: The concepts and philosophies of HTS can be productively applied to the discovery of new biopharmaceuticals. It is now possible, comprehensively and systematically, to enumerate, clone, produce and screen all secreted proteins, by building upon knowledge accumulated over the past two decades in HTS, genomics and parallel protein expression technologies. Each of the crucial operational components (comprehensive and high-quality cDNA library construction, proper protein-sequence classification, high-throughput protein production, medically relevant assays, state-of-the-art screening and data management) must be optimized to increase the chances of success. In this review, we draw comparisons between small-molecule and protein screening to illuminate common underlying principles as well as differences between the two operations.
Drug Discovery Today 09/2006; 11(15-16):718-24. · 6.83 Impact Factor
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Paul M Steed,
Malú G Tansey,
Jonathan Zalevsky,
Eugene A Zhukovsky,
John R Desjarlais,
David E Szymkowski,
Christina Abbott,
David Carmichael,
Cheryl Chan,
Lisa Cherry, [......],
Sabrina P Martinez,
Umesh S Muchhal,
Duc-Hanh T Nguyen,
Christopher O'Brien,
Donald O'Keefe,
Karen Singer,
Omid Vafa,
Jost Vielmetter,
Sean C Yoder,
Bassil I Dahiyat
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ABSTRACT: Tumor necrosis factor (TNF) is a key regulator of inflammatory responses and has been implicated in many pathological conditions. We used structure-based design to engineer variant TNF proteins that rapidly form heterotrimers with native TNF to give complexes that neither bind to nor stimulate signaling through TNF receptors. Thus, TNF is inactivated by sequestration. Dominant-negative TNFs represent a possible approach to anti-inflammatory biotherapeutics, and experiments in animal models show that the strategy can attenuate TNF-mediated pathology. Similar rational design could be used to engineer inhibitors of additional TNF superfamily cytokines as well as other multimeric ligands.
Science 10/2003; 301(5641):1895-8. · 31.20 Impact Factor
