A multistep high-content screening approach to identify novel functionally relevant target genes in pancreatic cancer
In order to foster the systematic identification of novel genes with important functional roles in pancreatic cancer, we have devised a multi-stage screening strategy to provide a rational basis for the selection of highly relevant novel candidate genes based on the results of functional high-conten...
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| Veröffentlicht in: | PloS one 2015-04, Vol.10 (4), p.e0122946-e0122946 |
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| creator | Buchholz, Malte Honstein, Tatjana Kirchhoff, Sandra Kreider, Ramona Schmidt, Harald Sipos, Bence Gress, Thomas M |
| description | In order to foster the systematic identification of novel genes with important functional roles in pancreatic cancer, we have devised a multi-stage screening strategy to provide a rational basis for the selection of highly relevant novel candidate genes based on the results of functional high-content analyses. The workflow comprised three consecutive stages: 1) serial gene expression profiling analyses of primary human pancreatic tissues as well as a number of in vivo and in vitro models of tumor-relevant characteristics in order to identify genes with conspicuous expression patterns; 2) use of 'reverse transfection array' technology for large-scale parallelized functional analyses of potential candidate genes in cell-based assays; and 3) selection of individual candidate genes for further in-depth examination of their cellular roles. A total of 14 genes, among them 8 from "druggable" gene families, were classified as high priority candidates for individual functional characterization. As an example to demonstrate the validity of the approach, comprehensive functional data on candidate gene ADRBK1/GRK2, which has previously not been implicated in pancreatic cancer, is presented. |
| doi_str_mv | 10.1371/journal.pone.0122946 |
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The workflow comprised three consecutive stages: 1) serial gene expression profiling analyses of primary human pancreatic tissues as well as a number of in vivo and in vitro models of tumor-relevant characteristics in order to identify genes with conspicuous expression patterns; 2) use of 'reverse transfection array' technology for large-scale parallelized functional analyses of potential candidate genes in cell-based assays; and 3) selection of individual candidate genes for further in-depth examination of their cellular roles. A total of 14 genes, among them 8 from "druggable" gene families, were classified as high priority candidates for individual functional characterization. As an example to demonstrate the validity of the approach, comprehensive functional data on candidate gene ADRBK1/GRK2, which has previously not been implicated in pancreatic cancer, is presented.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0122946</identifier><identifier>PMID: 25849100</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adult ; Apoptosis ; Arrays ; b-Adrenergic-receptor kinase ; Cancer ; Cancer genetics ; Cancer therapies ; Cell Line, Tumor ; Cell Proliferation ; Chemotherapy ; Disease ; Endocrinology ; Fetus ; G-Protein-Coupled Receptor Kinase 2 - genetics ; G-Protein-Coupled Receptor Kinase 2 - metabolism ; Gastroenterology ; Gene expression ; Gene Expression Profiling - methods ; Gene families ; Genes ; Genetic aspects ; Humans ; Hybridization ; Infection ; Intracellular Space - metabolism ; Kinases ; Leukemia ; Medical screening ; Metabolism ; Metastasis ; Pancreatic cancer ; Pancreatic Neoplasms - genetics ; Pancreatic Neoplasms - pathology ; Pancreatitis ; Protein Transport ; Proteins ; Proto-Oncogene Proteins c-crk - genetics ; Proto-Oncogene Proteins c-crk - metabolism ; Reverse Transcription ; Studies ; Target recognition ; Technology application ; Transfection ; Tumors ; Workflow</subject><ispartof>PloS one, 2015-04, Vol.10 (4), p.e0122946-e0122946</ispartof><rights>COPYRIGHT 2015 Public Library of Science</rights><rights>2015 Buchholz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2015 Buchholz et al 2015 Buchholz et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-f382c30cb7ad5bef4a82f88f1083d957e559b6f5462e4d28d0393121faf7b59f3</citedby><cites>FETCH-LOGICAL-c692t-f382c30cb7ad5bef4a82f88f1083d957e559b6f5462e4d28d0393121faf7b59f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4388713/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4388713/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79343,79344</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25849100$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Buchholz, Malte</creatorcontrib><creatorcontrib>Honstein, Tatjana</creatorcontrib><creatorcontrib>Kirchhoff, Sandra</creatorcontrib><creatorcontrib>Kreider, Ramona</creatorcontrib><creatorcontrib>Schmidt, Harald</creatorcontrib><creatorcontrib>Sipos, Bence</creatorcontrib><creatorcontrib>Gress, Thomas M</creatorcontrib><title>A multistep high-content screening approach to identify novel functionally relevant target genes in pancreatic cancer</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>In order to foster the systematic identification of novel genes with important functional roles in pancreatic cancer, we have devised a multi-stage screening strategy to provide a rational basis for the selection of highly relevant novel candidate genes based on the results of functional high-content analyses. 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| subjects | Adult Apoptosis Arrays b-Adrenergic-receptor kinase Cancer Cancer genetics Cancer therapies Cell Line, Tumor Cell Proliferation Chemotherapy Disease Endocrinology Fetus G-Protein-Coupled Receptor Kinase 2 - genetics G-Protein-Coupled Receptor Kinase 2 - metabolism Gastroenterology Gene expression Gene Expression Profiling - methods Gene families Genes Genetic aspects Humans Hybridization Infection Intracellular Space - metabolism Kinases Leukemia Medical screening Metabolism Metastasis Pancreatic cancer Pancreatic Neoplasms - genetics Pancreatic Neoplasms - pathology Pancreatitis Protein Transport Proteins Proto-Oncogene Proteins c-crk - genetics Proto-Oncogene Proteins c-crk - metabolism Reverse Transcription Studies Target recognition Technology application Transfection Tumors Workflow |
| title | A multistep high-content screening approach to identify novel functionally relevant target genes in pancreatic cancer |
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