Targeting polyIC to EGFR over-expressing cells using a dsRNA binding protein domain tethered to EGF.
Selective delivery of drugs to tumor cells can increase potency and reduce toxicity. In this study, we describe a novel recombinant chimeric protein, dsRBEC, which can bind polyIC and deliver it selectively into EGFR over-expressing tumor cells. dsRBEC, comprises the dsRNA binding domain (dsRBD) of human PKR (hPKR), which serves as the polyIC binding moiety, fused to human EGF (hEGF), the targeting moiety. dsRBEC shows high affinity towards EGFR and triggers ligand-induced endocytosis of the receptor, thus leading to the selective internalization of polyIC into EGFR over-expressing tumor cells. The targeted delivery of polyIC by dsRBEC induced cellular apoptosis and the secretion of IFN-β and other pro-inflammatory cytokines. dsRBEC-delivered polyIC is much more potent than naked polyIC and is expected to reduce the toxicity caused by systemic delivery of polyIC.
Unit of Cellular Signaling, Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.
Nufar
N
Edinger
Protein Purification Unit, Wolfson Center for Applied Structural Biology, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.
Mario
M
Lebendiker
Unit of Cellular Signaling, Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.
Shoshana
S
Klein
Unit of Cellular Signaling, Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.
Maya
M
Zigler
Unit of Cellular Signaling, Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.
Yael
Y
Langut
Unit of Cellular Signaling, Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.
Alexander
A
Levitzki
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D018946:Chemokine CCL5 / Q000096:biosynthesis / Q000378:metabolism; D003001:Cloning, Molecular; D016503:Drug Delivery Systems; D004705:Endocytosis; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007369:Interferon Inducers / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D016899:Interferon-beta / Q000096:biosynthesis / Q000378:metabolism; D061986:MCF-7 Cells; D011070:Poly I-C / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011485:Protein Binding; D000072417:Protein Domains; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000378:metabolism; D019892:eIF-2 Kinase / Q000235:genetics / Q000378:metabolism
C483024:CCL5 protein, human; D018946:Chemokine CCL5; D007369:Interferon Inducers; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D004815:Epidermal Growth Factor; D016899:Interferon-beta; C512478:EGFR protein, human; D066246:ErbB Receptors; C000602769:EIF2AK2 protein, human; D019892:eIF-2 Kinase; D011070:Poly I-C