Document Detail


Functional proteomic screen identifies a modulating role for CD44 in death receptor-mediated apoptosis.
MedLine Citation:
PMID:  15753387     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Apoptotic evasion is a hallmark of cancer and its resistance to chemotherapeutic drugs. Identification of cellular proteins that mediate apoptotic programs is a critical step toward the development of therapeutics aimed at overcoming apoptosis resistance. We developed an innovative high-throughput screen to identify proteins that modulate Fas ligand-mediated apoptosis using fluorophore-assisted light inactivation (HTS-FALIpop). The FALI protein knockdown strategy was coupled to a caspase activity assay with the ability to detect both proapoptotic and antiapoptotic surface molecules expressed by HT-1080 human fibrosarcoma cells. FALI of the Fas receptor (Fas/CD95) using a fluorescein-conjugated anti-Fas antibody abrogated Fas ligand-mediated caspase activation. Ninety-six single-chain variable fragment antibodies (scFv), selected for binding to the surface of HT-1080 cells, were screened by HTS-FALIpop. Three of the scFvs caused decreases in caspase induction after FALI of their protein targets. One of the targets of these positive scFvs was identified as CD44 and was validated by performing FALI using a CD44-specific monoclonal antibody, which resulted in similar protection from Fas apoptosis. CD44-targeted FALI was antiapoptotic in multiple human cancer cell lines, including both Fas signaling type I and II cells, and was also protective against other ligands of the tumor necrosis factor death receptor family. FALI of CD44 inhibited formation and activation of the death-inducing signaling complex, suggesting that CD44 regulates Fas at the cell surface. This mechanism of death receptor regulation represents a novel means of apoptosis modulation that could be exploited by pharmacologic agents.
Authors:
Robert S Hauptschein; Kevin E Sloan; Claudia Torella; Roya Moezzifard; Maryann Giel-Moloney; Carol Zehetmeier; Christine Unger; Leodevico L Ilag; Daniel G Jay
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Cancer research     Volume:  65     ISSN:  0008-5472     ISO Abbreviation:  Cancer Res.     Publication Date:  2005 Mar 
Date Detail:
Created Date:  2005-03-08     Completed Date:  2005-04-19     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2984705R     Medline TA:  Cancer Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1887-96     Citation Subset:  IM    
Affiliation:
Department of Physiology, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antigens, CD44 / genetics,  metabolism*
Antigens, CD95 / genetics,  metabolism*
Apoptosis*
Carcinoma, Renal Cell / metabolism,  pathology
Caspases / metabolism
Death Domain Receptor Signaling Adaptor Proteins
Enzyme Activation
Fibrosarcoma / metabolism,  pathology
Humans
Immunoglobulin Fragments / immunology*
Kidney Neoplasms / metabolism,  pathology
Lymphoma, T-Cell / metabolism,  pathology
Mammary Neoplasms, Experimental / metabolism,  pathology
Mass Spectrometry
Mice
Proteomics*
Receptors, Tumor Necrosis Factor / metabolism*
Tumor Cells, Cultured
Grant Support
ID/Acronym/Agency:
CA-81668/CA/NCI NIH HHS; DK-07542/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Antigens, CD44; 0/Antigens, CD95; 0/Death Domain Receptor Signaling Adaptor Proteins; 0/Immunoglobulin Fragments; 0/Receptors, Tumor Necrosis Factor; EC 3.4.22.-/Caspases

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