Screening compounds with a novel high-throughput ABCB1-mediated efflux assay identifies drugs with known therapeutic targets at risk for multidrug resistance interference
ABCB1, also known as P-glycoprotein (P-gp) or multidrug resistance protein 1 (MDR1), is a membrane-associated multidrug transporter of the ATP-binding cassette (ABC) transporter family. It is one of the most widely studied transporters that enable cancer cells to develop drug resistance. Reliable hi...
Gespeichert in:
| Veröffentlicht in: | PloS one 2013-04, Vol.8 (4), p.e60334 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | |
|---|---|
| container_issue | 4 |
| container_start_page | e60334 |
| container_title | PloS one |
| container_volume | 8 |
| creator | Ansbro, Megan R Shukla, Suneet Ambudkar, Suresh V Yuspa, Stuart H Li, Luowei |
| description | ABCB1, also known as P-glycoprotein (P-gp) or multidrug resistance protein 1 (MDR1), is a membrane-associated multidrug transporter of the ATP-binding cassette (ABC) transporter family. It is one of the most widely studied transporters that enable cancer cells to develop drug resistance. Reliable high-throughput assays that can identify compounds that interact with ABCB1 are crucial for developing new therapeutic drugs. A high-throughput assay for measuring ABCB1-mediated calcein AM efflux was developed using a fluorescent and phase-contrast live cell imaging system. This assay demonstrated the time- and dose-dependent accumulation of fluorescent calcein in ABCB1-overexpressing KB-V1 cells. Validation of the assay was performed with known ABCB1 inhibitors, XR9576, verapamil, and cyclosporin A, all of which displayed dose-dependent inhibition of ABCB1-mediated calcein AM efflux in this assay. Phase-contrast and fluorescent images taken by the imaging system provided additional opportunities for evaluating compounds that are cytotoxic or produce false positive signals. Compounds with known therapeutic targets and a kinase inhibitor library were screened. The assay identified multiple agents as inhibitors of ABCB1-mediated efflux and is highly reproducible. Among compounds identified as ABCB1 inhibitors, BEZ235, BI 2536, IKK 16, and ispinesib were further evaluated. The four compounds inhibited calcein AM efflux in a dose-dependent manner and were also active in the flow cytometry-based calcein AM efflux assay. BEZ235, BI 2536, and IKK 16 also successfully inhibited the labeling of ABCB1 with radiolabeled photoaffinity substrate [(125)I]iodoarylazidoprazosin. Inhibition of ABCB1 with XR9576 and cyclosporin A enhanced the cytotoxicity of BI 2536 to ABCB1-overexpressing cancer cells, HCT-15-Pgp, and decreased the IC50 value of BI 2536 by several orders of magnitude. This efficient, reliable, and simple high-throughput assay has identified ABCB1 substrates/inhibitors that may influence drug potency or drug-drug interactions and predict multidrug resistance in clinical treatment. |
| doi_str_mv | 10.1371/journal.pone.0060334 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_plos_</sourceid><recordid>TN_cdi_plos_journals_1330894738</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A478345690</galeid><doaj_id>oai_doaj_org_article_4b16bb4c8b154195b904b52036ec1831</doaj_id><sourcerecordid>A478345690</sourcerecordid><originalsourceid>FETCH-LOGICAL-c692t-242b61ab972418c525246627a420a2aee006d1363b7dd14546c7d5b13f186eec3</originalsourceid><addsrcrecordid>eNqNk1uL1DAYhoso7rr6D0QDguDFjM2haXsjzA4eBhYWXPU2pOnXNrudpCbpHv6Sv9KM012moCC5SJo879vkTb4keYnTJaY5fn9pR2dkvxysgWWa8pRS9ig5xiUlC05S-vhgfJQ88_4yTTNacP40OSI0Kyku-XHy60I5AKNNi5TdDnY0tUc3OnRIImOvoUedbrtF6Jwd224YA1qdrk_xYgu1lgFqBE3Tj7dIei_vkK7BBN1o8Kh2Yzs5XRl7Y1DowMkBxqAVCtK1EDySATntr1BjHdqOfdA7FXLgtQ_SKEDaBHANOIgfz5Mnjew9vJj6k-T7p4_f1l8WZ-efN-vV2ULxkoQFYaTiWFZlThguVEYywjgnuWQklUQCxKxqTDmt8rrGLGNc5XVWYdrgggMoepK83vsOvfViitkLTGlalCynRSQ2e6K28lIMTm-luxNWavFnwrpWSBfP2YNgFeZVxVRR4YzhMqvKlFVZvBIOChcUR68P09_GKoaqYoBO9jPT-YrRnWjttaCcEJ7TaPBmMnD25wg-_GPLE9XKuCttGhvN1FZ7JVYsLyjLeJlGavkXKrYatlrFh9boOD8TvJsJIhPgNrRy9F5sLr7-P3v-Y86-PWA7kH3ovO3j47HGz0G2B5Wz3jtoHpLDqdjVyX0aYlcnYqqTKHt1mPqD6L4w6G97JxCT</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1330894738</pqid></control><display><type>article</type><title>Screening compounds with a novel high-throughput ABCB1-mediated efflux assay identifies drugs with known therapeutic targets at risk for multidrug resistance interference</title><source>MEDLINE</source><source>DOAJ Directory of Open Access Journals</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><source>Public Library of Science (PLoS)</source><creator>Ansbro, Megan R ; Shukla, Suneet ; Ambudkar, Suresh V ; Yuspa, Stuart H ; Li, Luowei</creator><creatorcontrib>Ansbro, Megan R ; Shukla, Suneet ; Ambudkar, Suresh V ; Yuspa, Stuart H ; Li, Luowei</creatorcontrib><description>ABCB1, also known as P-glycoprotein (P-gp) or multidrug resistance protein 1 (MDR1), is a membrane-associated multidrug transporter of the ATP-binding cassette (ABC) transporter family. It is one of the most widely studied transporters that enable cancer cells to develop drug resistance. Reliable high-throughput assays that can identify compounds that interact with ABCB1 are crucial for developing new therapeutic drugs. A high-throughput assay for measuring ABCB1-mediated calcein AM efflux was developed using a fluorescent and phase-contrast live cell imaging system. This assay demonstrated the time- and dose-dependent accumulation of fluorescent calcein in ABCB1-overexpressing KB-V1 cells. Validation of the assay was performed with known ABCB1 inhibitors, XR9576, verapamil, and cyclosporin A, all of which displayed dose-dependent inhibition of ABCB1-mediated calcein AM efflux in this assay. Phase-contrast and fluorescent images taken by the imaging system provided additional opportunities for evaluating compounds that are cytotoxic or produce false positive signals. Compounds with known therapeutic targets and a kinase inhibitor library were screened. The assay identified multiple agents as inhibitors of ABCB1-mediated efflux and is highly reproducible. Among compounds identified as ABCB1 inhibitors, BEZ235, BI 2536, IKK 16, and ispinesib were further evaluated. The four compounds inhibited calcein AM efflux in a dose-dependent manner and were also active in the flow cytometry-based calcein AM efflux assay. BEZ235, BI 2536, and IKK 16 also successfully inhibited the labeling of ABCB1 with radiolabeled photoaffinity substrate [(125)I]iodoarylazidoprazosin. Inhibition of ABCB1 with XR9576 and cyclosporin A enhanced the cytotoxicity of BI 2536 to ABCB1-overexpressing cancer cells, HCT-15-Pgp, and decreased the IC50 value of BI 2536 by several orders of magnitude. This efficient, reliable, and simple high-throughput assay has identified ABCB1 substrates/inhibitors that may influence drug potency or drug-drug interactions and predict multidrug resistance in clinical treatment.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0060334</identifier><identifier>PMID: 23593196</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Assaying ; ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 - antagonists & inhibitors ; ATP-Binding Cassette, Sub-Family B, Member 1 - physiology ; Binding sites ; Biology ; Calcein ; Cancer ; Cancer therapies ; Cell Line ; Chemical compounds ; Cyclosporin A ; Cyclosporine - pharmacology ; Cytometry ; Cytotoxicity ; Drug development ; Drug resistance ; Drug Resistance, Multiple ; Drugs ; Efflux ; Enzyme inhibitors ; Flow cytometry ; Fluorescence ; Glycoproteins ; Health aspects ; High-Throughput Screening Assays ; Humans ; Image contrast ; Inhibitors ; Kinases ; Laboratories ; Libraries ; MDR1 protein ; Medical research ; Medical screening ; Medicine ; Microbial drug resistance ; Multidrug resistance ; Multidrug resistant organisms ; P-Glycoprotein ; Pharmacology ; Proteins ; Quinolines - pharmacology ; Reproducibility of Results ; Substrate inhibition ; Substrates ; Toxicity ; Verapamil ; Verapamil - pharmacology</subject><ispartof>PloS one, 2013-04, Vol.8 (4), p.e60334</ispartof><rights>COPYRIGHT 2013 Public Library of Science</rights><rights>2013. This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2013</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-242b61ab972418c525246627a420a2aee006d1363b7dd14546c7d5b13f186eec3</citedby><cites>FETCH-LOGICAL-c692t-242b61ab972418c525246627a420a2aee006d1363b7dd14546c7d5b13f186eec3</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/PMC3622673/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3622673/$$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/23593196$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ansbro, Megan R</creatorcontrib><creatorcontrib>Shukla, Suneet</creatorcontrib><creatorcontrib>Ambudkar, Suresh V</creatorcontrib><creatorcontrib>Yuspa, Stuart H</creatorcontrib><creatorcontrib>Li, Luowei</creatorcontrib><title>Screening compounds with a novel high-throughput ABCB1-mediated efflux assay identifies drugs with known therapeutic targets at risk for multidrug resistance interference</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>ABCB1, also known as P-glycoprotein (P-gp) or multidrug resistance protein 1 (MDR1), is a membrane-associated multidrug transporter of the ATP-binding cassette (ABC) transporter family. It is one of the most widely studied transporters that enable cancer cells to develop drug resistance. Reliable high-throughput assays that can identify compounds that interact with ABCB1 are crucial for developing new therapeutic drugs. A high-throughput assay for measuring ABCB1-mediated calcein AM efflux was developed using a fluorescent and phase-contrast live cell imaging system. This assay demonstrated the time- and dose-dependent accumulation of fluorescent calcein in ABCB1-overexpressing KB-V1 cells. Validation of the assay was performed with known ABCB1 inhibitors, XR9576, verapamil, and cyclosporin A, all of which displayed dose-dependent inhibition of ABCB1-mediated calcein AM efflux in this assay. Phase-contrast and fluorescent images taken by the imaging system provided additional opportunities for evaluating compounds that are cytotoxic or produce false positive signals. Compounds with known therapeutic targets and a kinase inhibitor library were screened. The assay identified multiple agents as inhibitors of ABCB1-mediated efflux and is highly reproducible. Among compounds identified as ABCB1 inhibitors, BEZ235, BI 2536, IKK 16, and ispinesib were further evaluated. The four compounds inhibited calcein AM efflux in a dose-dependent manner and were also active in the flow cytometry-based calcein AM efflux assay. BEZ235, BI 2536, and IKK 16 also successfully inhibited the labeling of ABCB1 with radiolabeled photoaffinity substrate [(125)I]iodoarylazidoprazosin. Inhibition of ABCB1 with XR9576 and cyclosporin A enhanced the cytotoxicity of BI 2536 to ABCB1-overexpressing cancer cells, HCT-15-Pgp, and decreased the IC50 value of BI 2536 by several orders of magnitude. This efficient, reliable, and simple high-throughput assay has identified ABCB1 substrates/inhibitors that may influence drug potency or drug-drug interactions and predict multidrug resistance in clinical treatment.</description><subject>Assaying</subject><subject>ATP Binding Cassette Transporter, Sub-Family B</subject><subject>ATP-Binding Cassette, Sub-Family B, Member 1 - antagonists & inhibitors</subject><subject>ATP-Binding Cassette, Sub-Family B, Member 1 - physiology</subject><subject>Binding sites</subject><subject>Biology</subject><subject>Calcein</subject><subject>Cancer</subject><subject>Cancer therapies</subject><subject>Cell Line</subject><subject>Chemical compounds</subject><subject>Cyclosporin A</subject><subject>Cyclosporine - pharmacology</subject><subject>Cytometry</subject><subject>Cytotoxicity</subject><subject>Drug development</subject><subject>Drug resistance</subject><subject>Drug Resistance, Multiple</subject><subject>Drugs</subject><subject>Efflux</subject><subject>Enzyme inhibitors</subject><subject>Flow cytometry</subject><subject>Fluorescence</subject><subject>Glycoproteins</subject><subject>Health aspects</subject><subject>High-Throughput Screening Assays</subject><subject>Humans</subject><subject>Image contrast</subject><subject>Inhibitors</subject><subject>Kinases</subject><subject>Laboratories</subject><subject>Libraries</subject><subject>MDR1 protein</subject><subject>Medical research</subject><subject>Medical screening</subject><subject>Medicine</subject><subject>Microbial drug resistance</subject><subject>Multidrug resistance</subject><subject>Multidrug resistant organisms</subject><subject>P-Glycoprotein</subject><subject>Pharmacology</subject><subject>Proteins</subject><subject>Quinolines - pharmacology</subject><subject>Reproducibility of Results</subject><subject>Substrate inhibition</subject><subject>Substrates</subject><subject>Toxicity</subject><subject>Verapamil</subject><subject>Verapamil - pharmacology</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk1uL1DAYhoso7rr6D0QDguDFjM2haXsjzA4eBhYWXPU2pOnXNrudpCbpHv6Sv9KM012moCC5SJo879vkTb4keYnTJaY5fn9pR2dkvxysgWWa8pRS9ig5xiUlC05S-vhgfJQ88_4yTTNacP40OSI0Kyku-XHy60I5AKNNi5TdDnY0tUc3OnRIImOvoUedbrtF6Jwd224YA1qdrk_xYgu1lgFqBE3Tj7dIei_vkK7BBN1o8Kh2Yzs5XRl7Y1DowMkBxqAVCtK1EDySATntr1BjHdqOfdA7FXLgtQ_SKEDaBHANOIgfz5Mnjew9vJj6k-T7p4_f1l8WZ-efN-vV2ULxkoQFYaTiWFZlThguVEYywjgnuWQklUQCxKxqTDmt8rrGLGNc5XVWYdrgggMoepK83vsOvfViitkLTGlalCynRSQ2e6K28lIMTm-luxNWavFnwrpWSBfP2YNgFeZVxVRR4YzhMqvKlFVZvBIOChcUR68P09_GKoaqYoBO9jPT-YrRnWjttaCcEJ7TaPBmMnD25wg-_GPLE9XKuCttGhvN1FZ7JVYsLyjLeJlGavkXKrYatlrFh9boOD8TvJsJIhPgNrRy9F5sLr7-P3v-Y86-PWA7kH3ovO3j47HGz0G2B5Wz3jtoHpLDqdjVyX0aYlcnYqqTKHt1mPqD6L4w6G97JxCT</recordid><startdate>20130410</startdate><enddate>20130410</enddate><creator>Ansbro, Megan R</creator><creator>Shukla, Suneet</creator><creator>Ambudkar, Suresh V</creator><creator>Yuspa, Stuart H</creator><creator>Li, Luowei</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20130410</creationdate><title>Screening compounds with a novel high-throughput ABCB1-mediated efflux assay identifies drugs with known therapeutic targets at risk for multidrug resistance interference</title><author>Ansbro, Megan R ; Shukla, Suneet ; Ambudkar, Suresh V ; Yuspa, Stuart H ; Li, Luowei</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-242b61ab972418c525246627a420a2aee006d1363b7dd14546c7d5b13f186eec3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Assaying</topic><topic>ATP Binding Cassette Transporter, Sub-Family B</topic><topic>ATP-Binding Cassette, Sub-Family B, Member 1 - antagonists & inhibitors</topic><topic>ATP-Binding Cassette, Sub-Family B, Member 1 - physiology</topic><topic>Binding sites</topic><topic>Biology</topic><topic>Calcein</topic><topic>Cancer</topic><topic>Cancer therapies</topic><topic>Cell Line</topic><topic>Chemical compounds</topic><topic>Cyclosporin A</topic><topic>Cyclosporine - pharmacology</topic><topic>Cytometry</topic><topic>Cytotoxicity</topic><topic>Drug development</topic><topic>Drug resistance</topic><topic>Drug Resistance, Multiple</topic><topic>Drugs</topic><topic>Efflux</topic><topic>Enzyme inhibitors</topic><topic>Flow cytometry</topic><topic>Fluorescence</topic><topic>Glycoproteins</topic><topic>Health aspects</topic><topic>High-Throughput Screening Assays</topic><topic>Humans</topic><topic>Image contrast</topic><topic>Inhibitors</topic><topic>Kinases</topic><topic>Laboratories</topic><topic>Libraries</topic><topic>MDR1 protein</topic><topic>Medical research</topic><topic>Medical screening</topic><topic>Medicine</topic><topic>Microbial drug resistance</topic><topic>Multidrug resistance</topic><topic>Multidrug resistant organisms</topic><topic>P-Glycoprotein</topic><topic>Pharmacology</topic><topic>Proteins</topic><topic>Quinolines - pharmacology</topic><topic>Reproducibility of Results</topic><topic>Substrate inhibition</topic><topic>Substrates</topic><topic>Toxicity</topic><topic>Verapamil</topic><topic>Verapamil - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ansbro, Megan R</creatorcontrib><creatorcontrib>Shukla, Suneet</creatorcontrib><creatorcontrib>Ambudkar, Suresh V</creatorcontrib><creatorcontrib>Yuspa, Stuart H</creatorcontrib><creatorcontrib>Li, Luowei</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - Academic</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials Science Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ansbro, Megan R</au><au>Shukla, Suneet</au><au>Ambudkar, Suresh V</au><au>Yuspa, Stuart H</au><au>Li, Luowei</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Screening compounds with a novel high-throughput ABCB1-mediated efflux assay identifies drugs with known therapeutic targets at risk for multidrug resistance interference</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-04-10</date><risdate>2013</risdate><volume>8</volume><issue>4</issue><spage>e60334</spage><pages>e60334-</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>ABCB1, also known as P-glycoprotein (P-gp) or multidrug resistance protein 1 (MDR1), is a membrane-associated multidrug transporter of the ATP-binding cassette (ABC) transporter family. It is one of the most widely studied transporters that enable cancer cells to develop drug resistance. Reliable high-throughput assays that can identify compounds that interact with ABCB1 are crucial for developing new therapeutic drugs. A high-throughput assay for measuring ABCB1-mediated calcein AM efflux was developed using a fluorescent and phase-contrast live cell imaging system. This assay demonstrated the time- and dose-dependent accumulation of fluorescent calcein in ABCB1-overexpressing KB-V1 cells. Validation of the assay was performed with known ABCB1 inhibitors, XR9576, verapamil, and cyclosporin A, all of which displayed dose-dependent inhibition of ABCB1-mediated calcein AM efflux in this assay. Phase-contrast and fluorescent images taken by the imaging system provided additional opportunities for evaluating compounds that are cytotoxic or produce false positive signals. Compounds with known therapeutic targets and a kinase inhibitor library were screened. The assay identified multiple agents as inhibitors of ABCB1-mediated efflux and is highly reproducible. Among compounds identified as ABCB1 inhibitors, BEZ235, BI 2536, IKK 16, and ispinesib were further evaluated. The four compounds inhibited calcein AM efflux in a dose-dependent manner and were also active in the flow cytometry-based calcein AM efflux assay. BEZ235, BI 2536, and IKK 16 also successfully inhibited the labeling of ABCB1 with radiolabeled photoaffinity substrate [(125)I]iodoarylazidoprazosin. Inhibition of ABCB1 with XR9576 and cyclosporin A enhanced the cytotoxicity of BI 2536 to ABCB1-overexpressing cancer cells, HCT-15-Pgp, and decreased the IC50 value of BI 2536 by several orders of magnitude. This efficient, reliable, and simple high-throughput assay has identified ABCB1 substrates/inhibitors that may influence drug potency or drug-drug interactions and predict multidrug resistance in clinical treatment.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23593196</pmid><doi>10.1371/journal.pone.0060334</doi><tpages>e60334</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2013-04, Vol.8 (4), p.e60334 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1330894738 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Assaying ATP Binding Cassette Transporter, Sub-Family B ATP-Binding Cassette, Sub-Family B, Member 1 - antagonists & inhibitors ATP-Binding Cassette, Sub-Family B, Member 1 - physiology Binding sites Biology Calcein Cancer Cancer therapies Cell Line Chemical compounds Cyclosporin A Cyclosporine - pharmacology Cytometry Cytotoxicity Drug development Drug resistance Drug Resistance, Multiple Drugs Efflux Enzyme inhibitors Flow cytometry Fluorescence Glycoproteins Health aspects High-Throughput Screening Assays Humans Image contrast Inhibitors Kinases Laboratories Libraries MDR1 protein Medical research Medical screening Medicine Microbial drug resistance Multidrug resistance Multidrug resistant organisms P-Glycoprotein Pharmacology Proteins Quinolines - pharmacology Reproducibility of Results Substrate inhibition Substrates Toxicity Verapamil Verapamil - pharmacology |
| title | Screening compounds with a novel high-throughput ABCB1-mediated efflux assay identifies drugs with known therapeutic targets at risk for multidrug resistance interference |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-01T18%3A31%3A53IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Screening%20compounds%20with%20a%20novel%20high-throughput%20ABCB1-mediated%20efflux%20assay%20identifies%20drugs%20with%20known%20therapeutic%20targets%20at%20risk%20for%20multidrug%20resistance%20interference&rft.jtitle=PloS%20one&rft.au=Ansbro,%20Megan%20R&rft.date=2013-04-10&rft.volume=8&rft.issue=4&rft.spage=e60334&rft.pages=e60334-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0060334&rft_dat=%3Cgale_plos_%3EA478345690%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1330894738&rft_id=info:pmid/23593196&rft_galeid=A478345690&rft_doaj_id=oai_doaj_org_article_4b16bb4c8b154195b904b52036ec1831&rfr_iscdi=true |