Identification of modulators of TRAIL-induced apoptosis via RNAi-based phenotypic screening.
New opportunities in mammalian functional genomics are emerging through the combination of high throughput technology and methods that allow manipulation of gene expression in living cells. Here we describe the application of an RNAi-based forward genomics approach toward understanding the biology and mechanism of TRAIL-induced apoptosis. TRAIL is a TNF superfamily member that induces selective cytotoxicity of tumor cells when bound to its cognate receptors. In addition to detecting well-characterized genes in the apoptosis pathway, we uncover several modulators including DOBI, a gene required for progression of the apoptotic signal through the intrinsic mitochondrial cell death pathway, and MIRSA, a gene that acts to limit TRAIL-induced apoptosis. Moreover, our data suggest a role for MYC and the WNT pathway in maintaining susceptibility to TRAIL. Collectively, these observations offer several insights on how TRAIL mediates the selective killing of tumor cells and demonstrate the utility of large-scale RNAi screens in mammalian cells.
Aza-Blanc
Pedro
P
Genomics Institute of the Novartis Research Foundation, 10675 John J. Hopkins Drive, San Diego, CA 92121, USA. aza-blanc@gnf.org
Cooper
Christopher L
CL
Wagner
Klaus
K
Batalov
Serge
S
Deveraux
Quinn L
QL
Cooke
Michael P
MP
D017209:Apoptosis / Q000235:genetics* / Q000276:immunology*; D051017:Apoptosis Regulatory Proteins; D005786:Gene Expression Regulation / Q000235:genetics; D005820:Genetic Testing / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D007157:Immunologic Surveillance / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000276:immunology*; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism; D009369:Neoplasms / Q000276:immunology*; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D034622:RNA Interference / Q000276:immunology*; D015398:Signal Transduction / Q000235:genetics; D053221:TNF-Related Apoptosis-Inducing Ligand; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*; D051153:Wnt Proteins; D029961:Zebrafish Proteins
D051017:Apoptosis Regulatory Proteins; D008562:Membrane Glycoproteins; D011518:Proto-Oncogene Proteins; D016271:Proto-Oncogene Proteins c-myc; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D014409:Tumor Necrosis Factor-alpha; D051153:Wnt Proteins; D029961:Zebrafish Proteins