Rescue of murine F508del CFTR activity in native intestine by low temperature and proteasome inhibitors
Most patients with Cystic Fibrosis (CF) carry at least one allele with the F508del mutation, resulting in a CFTR chloride channel protein with a processing, gating and stability defect, but with substantial residual activity when correctly sorted to the apical membranes of epithelial cells. New ther...
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| description | Most patients with Cystic Fibrosis (CF) carry at least one allele with the F508del mutation, resulting in a CFTR chloride channel protein with a processing, gating and stability defect, but with substantial residual activity when correctly sorted to the apical membranes of epithelial cells. New therapies are therefore aimed at improving the folding and trafficking of F508del CFTR, (CFTR correctors) or at enhancing the open probability of the CFTR chloride channel (CFTR potentiators). Preventing premature breakdown of F508del CFTR is an alternative or additional strategy, which is investigated in this study. We established an ex vivo assay for murine F508del CFTR rescue in native intestinal epithelium that can be used as a pre-clinical test for candidate therapeutics. Overnight incubation of muscle stripped ileum in modified William's E medium at low temperature (26°C), and 4 h or 6 h incubation at 37°C with different proteasome inhibitors (PI: ALLN, MG-132, epoxomicin, PS341/bortezomib) resulted in fifty to hundred percent respectively of the wild type CFTR mediated chloride secretion (forskolin induced short-circuit current). The functional rescue was accompanied by enhanced expression of the murine F508del CFTR protein at the apical surface of intestinal crypts and a gain in the amount of complex-glycosylated CFTR (band C) up to 20% of WT levels. Sustained rescue in the presence of brefeldin A shows the involvement of a post-Golgi compartment in murine F508del CFTR degradation, as was shown earlier for its human counterpart. Our data show that proteasome inhibitors are promising candidate compounds for improving rescue of human F508del CFTR function, in combination with available correctors and potentiators. |
| doi_str_mv | 10.1371/journal.pone.0052070 |
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New therapies are therefore aimed at improving the folding and trafficking of F508del CFTR, (CFTR correctors) or at enhancing the open probability of the CFTR chloride channel (CFTR potentiators). Preventing premature breakdown of F508del CFTR is an alternative or additional strategy, which is investigated in this study. We established an ex vivo assay for murine F508del CFTR rescue in native intestinal epithelium that can be used as a pre-clinical test for candidate therapeutics. Overnight incubation of muscle stripped ileum in modified William's E medium at low temperature (26°C), and 4 h or 6 h incubation at 37°C with different proteasome inhibitors (PI: ALLN, MG-132, epoxomicin, PS341/bortezomib) resulted in fifty to hundred percent respectively of the wild type CFTR mediated chloride secretion (forskolin induced short-circuit current). The functional rescue was accompanied by enhanced expression of the murine F508del CFTR protein at the apical surface of intestinal crypts and a gain in the amount of complex-glycosylated CFTR (band C) up to 20% of WT levels. Sustained rescue in the presence of brefeldin A shows the involvement of a post-Golgi compartment in murine F508del CFTR degradation, as was shown earlier for its human counterpart. Our data show that proteasome inhibitors are promising candidate compounds for improving rescue of human F508del CFTR function, in combination with available correctors and potentiators.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0052070</identifier><identifier>PMID: 23284872</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Animals ; Autophagy ; Biological Transport - drug effects ; Biology ; Bortezomib ; Brefeldin A ; C band ; Cancer ; Cell culture ; Cell cycle ; Channel gating ; Chloride ; Chlorides ; Chlorides - metabolism ; Cold Temperature ; Consortia ; Crypts ; Cystic fibrosis ; Cystic Fibrosis - genetics ; Cystic Fibrosis - metabolism ; Cystic fibrosis transmembrane conductance regulator ; Cystic Fibrosis Transmembrane Conductance Regulator - genetics ; Cystic Fibrosis Transmembrane Conductance Regulator - metabolism ; Defects ; Epithelial cells ; Epithelium ; Fibrosis ; Forskolin ; Gastroenterology ; Golgi apparatus ; Ileum ; Incubation ; Inhibitors ; Intestinal Mucosa - drug effects ; Intestinal Mucosa - metabolism ; Intestine ; Intestines - drug effects ; Intestines - metabolism ; Laboratory animals ; Low temperature ; Medicine ; Membranes ; Mice ; Mice, Knockout ; Multiple myeloma ; Muscles ; Mutation ; Proteasome inhibitors ; Proteasome Inhibitors - pharmacology ; Proteins ; Short circuits ; Short-circuit current</subject><ispartof>PloS one, 2012-12, Vol.7 (12), p.e52070-e52070</ispartof><rights>COPYRIGHT 2012 Public Library of Science</rights><rights>2012 Wilke et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://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>2012 Wilke et al 2012 Wilke et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-bf6083e9d830eec8df707eba79283b8fa383ecc0eb8c67566160e014715df6983</citedby><cites>FETCH-LOGICAL-c758t-bf6083e9d830eec8df707eba79283b8fa383ecc0eb8c67566160e014715df6983</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/PMC3528711/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3528711/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23284872$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Vij, Neeraj</contributor><creatorcontrib>Wilke, Martina</creatorcontrib><creatorcontrib>Bot, Alice</creatorcontrib><creatorcontrib>Jorna, Huub</creatorcontrib><creatorcontrib>Scholte, Bob J</creatorcontrib><creatorcontrib>de Jonge, Hugo R</creatorcontrib><title>Rescue of murine F508del CFTR activity in native intestine by low temperature and proteasome inhibitors</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Most patients with Cystic Fibrosis (CF) carry at least one allele with the F508del mutation, resulting in a CFTR chloride channel protein with a processing, gating and stability defect, but with substantial residual activity when correctly sorted to the apical membranes of epithelial cells. 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The functional rescue was accompanied by enhanced expression of the murine F508del CFTR protein at the apical surface of intestinal crypts and a gain in the amount of complex-glycosylated CFTR (band C) up to 20% of WT levels. Sustained rescue in the presence of brefeldin A shows the involvement of a post-Golgi compartment in murine F508del CFTR degradation, as was shown earlier for its human counterpart. Our data show that proteasome inhibitors are promising candidate compounds for improving rescue of human F508del CFTR function, in combination with available correctors and potentiators.</description><subject>Animals</subject><subject>Autophagy</subject><subject>Biological Transport - drug effects</subject><subject>Biology</subject><subject>Bortezomib</subject><subject>Brefeldin A</subject><subject>C band</subject><subject>Cancer</subject><subject>Cell culture</subject><subject>Cell cycle</subject><subject>Channel gating</subject><subject>Chloride</subject><subject>Chlorides</subject><subject>Chlorides - metabolism</subject><subject>Cold Temperature</subject><subject>Consortia</subject><subject>Crypts</subject><subject>Cystic fibrosis</subject><subject>Cystic Fibrosis - genetics</subject><subject>Cystic Fibrosis - metabolism</subject><subject>Cystic fibrosis transmembrane conductance regulator</subject><subject>Cystic Fibrosis Transmembrane Conductance Regulator - genetics</subject><subject>Cystic Fibrosis Transmembrane Conductance Regulator - metabolism</subject><subject>Defects</subject><subject>Epithelial cells</subject><subject>Epithelium</subject><subject>Fibrosis</subject><subject>Forskolin</subject><subject>Gastroenterology</subject><subject>Golgi apparatus</subject><subject>Ileum</subject><subject>Incubation</subject><subject>Inhibitors</subject><subject>Intestinal Mucosa - drug effects</subject><subject>Intestinal Mucosa - metabolism</subject><subject>Intestine</subject><subject>Intestines - drug effects</subject><subject>Intestines - metabolism</subject><subject>Laboratory animals</subject><subject>Low temperature</subject><subject>Medicine</subject><subject>Membranes</subject><subject>Mice</subject><subject>Mice, Knockout</subject><subject>Multiple myeloma</subject><subject>Muscles</subject><subject>Mutation</subject><subject>Proteasome inhibitors</subject><subject>Proteasome Inhibitors - pharmacology</subject><subject>Proteins</subject><subject>Short circuits</subject><subject>Short-circuit current</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNk9-LEzEQxxdRvPP0PxBdEEQfWvNjN8m-CEexWjg4qKevIcnOtim7m16SPe1_b2r3jlbuQfKQYeYz30kmmSx7jdEUU44_bdzge9VOt66HKUIlQRw9yc5xRcmEEUSfHtln2YsQNgmigrHn2RmhRBSCk_NstYRgBshdk3eDtz3k8xKJGtp8Nr9Z5spEe2fjLrd93qtkQ7IihLgn9S5v3a88QrcFr-LgIVd9nW-9i6CC6_bs2mobnQ8vs2eNagO8GveL7Mf8y83s2-Tq-utidnk1MbwUcaIbhgSFqhYUARhRNxxx0IpXRFAtGkVT1BgEWhjGS8YwQ4BwwXFZN6wS9CJ7e9Ddti7IsUdBYko4LhgqaCIWB6J2aiO33nbK76RTVv51OL-SykdrWpBQKE6UKIluREFEVdUVr4lm2pS60VWTtD6P1QbdQW2gj161J6Knkd6u5crdSVoSwTFOAh9GAe9uh9RX2dlgoG1VD25I5yacYk4QIwl99w_6-O1GaqXSBWzfuFTX7EXlZcE5KkvOeKKmj1Bp1dBZkz5UY5P_JOHjSUJiIvyOKzWEIBffl__PXv88Zd8fsWtQbVwH1w7Ruj6cgsUBNN6F4KF5aDJGcj8P992Q-3mQ4zyktDfHD_SQdD8A9A-JLgWb</recordid><startdate>20121221</startdate><enddate>20121221</enddate><creator>Wilke, Martina</creator><creator>Bot, Alice</creator><creator>Jorna, Huub</creator><creator>Scholte, Bob J</creator><creator>de Jonge, Hugo R</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>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>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20121221</creationdate><title>Rescue of murine F508del CFTR activity in native intestine by low temperature and proteasome inhibitors</title><author>Wilke, Martina ; Bot, Alice ; Jorna, Huub ; Scholte, Bob J ; de Jonge, Hugo R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c758t-bf6083e9d830eec8df707eba79283b8fa383ecc0eb8c67566160e014715df6983</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Animals</topic><topic>Autophagy</topic><topic>Biological Transport - drug effects</topic><topic>Biology</topic><topic>Bortezomib</topic><topic>Brefeldin A</topic><topic>C band</topic><topic>Cancer</topic><topic>Cell culture</topic><topic>Cell cycle</topic><topic>Channel gating</topic><topic>Chloride</topic><topic>Chlorides</topic><topic>Chlorides - metabolism</topic><topic>Cold Temperature</topic><topic>Consortia</topic><topic>Crypts</topic><topic>Cystic fibrosis</topic><topic>Cystic Fibrosis - genetics</topic><topic>Cystic Fibrosis - metabolism</topic><topic>Cystic fibrosis transmembrane conductance regulator</topic><topic>Cystic Fibrosis Transmembrane Conductance Regulator - genetics</topic><topic>Cystic Fibrosis Transmembrane Conductance Regulator - metabolism</topic><topic>Defects</topic><topic>Epithelial cells</topic><topic>Epithelium</topic><topic>Fibrosis</topic><topic>Forskolin</topic><topic>Gastroenterology</topic><topic>Golgi apparatus</topic><topic>Ileum</topic><topic>Incubation</topic><topic>Inhibitors</topic><topic>Intestinal Mucosa - drug effects</topic><topic>Intestinal Mucosa - 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Academic</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>Wilke, Martina</au><au>Bot, Alice</au><au>Jorna, Huub</au><au>Scholte, Bob J</au><au>de Jonge, Hugo R</au><au>Vij, Neeraj</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rescue of murine F508del CFTR activity in native intestine by low temperature and proteasome inhibitors</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2012-12-21</date><risdate>2012</risdate><volume>7</volume><issue>12</issue><spage>e52070</spage><epage>e52070</epage><pages>e52070-e52070</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Most patients with Cystic Fibrosis (CF) carry at least one allele with the F508del mutation, resulting in a CFTR chloride channel protein with a processing, gating and stability defect, but with substantial residual activity when correctly sorted to the apical membranes of epithelial cells. New therapies are therefore aimed at improving the folding and trafficking of F508del CFTR, (CFTR correctors) or at enhancing the open probability of the CFTR chloride channel (CFTR potentiators). Preventing premature breakdown of F508del CFTR is an alternative or additional strategy, which is investigated in this study. We established an ex vivo assay for murine F508del CFTR rescue in native intestinal epithelium that can be used as a pre-clinical test for candidate therapeutics. Overnight incubation of muscle stripped ileum in modified William's E medium at low temperature (26°C), and 4 h or 6 h incubation at 37°C with different proteasome inhibitors (PI: ALLN, MG-132, epoxomicin, PS341/bortezomib) resulted in fifty to hundred percent respectively of the wild type CFTR mediated chloride secretion (forskolin induced short-circuit current). The functional rescue was accompanied by enhanced expression of the murine F508del CFTR protein at the apical surface of intestinal crypts and a gain in the amount of complex-glycosylated CFTR (band C) up to 20% of WT levels. Sustained rescue in the presence of brefeldin A shows the involvement of a post-Golgi compartment in murine F508del CFTR degradation, as was shown earlier for its human counterpart. Our data show that proteasome inhibitors are promising candidate compounds for improving rescue of human F508del CFTR function, in combination with available correctors and potentiators.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23284872</pmid><doi>10.1371/journal.pone.0052070</doi><tpages>e52070</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Autophagy Biological Transport - drug effects Biology Bortezomib Brefeldin A C band Cancer Cell culture Cell cycle Channel gating Chloride Chlorides Chlorides - metabolism Cold Temperature Consortia Crypts Cystic fibrosis Cystic Fibrosis - genetics Cystic Fibrosis - metabolism Cystic fibrosis transmembrane conductance regulator Cystic Fibrosis Transmembrane Conductance Regulator - genetics Cystic Fibrosis Transmembrane Conductance Regulator - metabolism Defects Epithelial cells Epithelium Fibrosis Forskolin Gastroenterology Golgi apparatus Ileum Incubation Inhibitors Intestinal Mucosa - drug effects Intestinal Mucosa - metabolism Intestine Intestines - drug effects Intestines - metabolism Laboratory animals Low temperature Medicine Membranes Mice Mice, Knockout Multiple myeloma Muscles Mutation Proteasome inhibitors Proteasome Inhibitors - pharmacology Proteins Short circuits Short-circuit current |
| title | Rescue of murine F508del CFTR activity in native intestine by low temperature and proteasome inhibitors |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-24T17%3A40%3A01IST&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=Rescue%20of%20murine%20F508del%20CFTR%20activity%20in%20native%20intestine%20by%20low%20temperature%20and%20proteasome%20inhibitors&rft.jtitle=PloS%20one&rft.au=Wilke,%20Martina&rft.date=2012-12-21&rft.volume=7&rft.issue=12&rft.spage=e52070&rft.epage=e52070&rft.pages=e52070-e52070&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0052070&rft_dat=%3Cgale_plos_%3EA477055767%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=1327146043&rft_id=info:pmid/23284872&rft_galeid=A477055767&rft_doaj_id=oai_doaj_org_article_e4a72a852bf842899d97d2b6bc5bfb9f&rfr_iscdi=true |