Circumventing furin enhances factor VIII biological activity and ameliorates bleeding phenotypes in hemophilia models

Processing by the proprotein convertase furin is believed to be critical for the biological activity of multiple proteins involved in hemostasis, including coagulation factor VIII (FVIII). This belief prompted the retention of the furin recognition motif (amino acids 1645-1648) in the design of B-do...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:JCI insight 2016-10, Vol.1 (16), p.e89371-e89371
Hauptverfasser: Siner, Joshua I, Samelson-Jones, Benjamin J, Crudele, Julie M, French, Robert A, Lee, Benjamin J, Zhou, Shanzhen, Merricks, Elizabeth, Raymer, Robin, Nichols, Timothy C, Camire, Rodney M, Arruda, Valder R
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page e89371
container_issue 16
container_start_page e89371
container_title JCI insight
container_volume 1
creator Siner, Joshua I
Samelson-Jones, Benjamin J
Crudele, Julie M
French, Robert A
Lee, Benjamin J
Zhou, Shanzhen
Merricks, Elizabeth
Raymer, Robin
Nichols, Timothy C
Camire, Rodney M
Arruda, Valder R
description Processing by the proprotein convertase furin is believed to be critical for the biological activity of multiple proteins involved in hemostasis, including coagulation factor VIII (FVIII). This belief prompted the retention of the furin recognition motif (amino acids 1645-1648) in the design of B-domain-deleted FVIII (FVIII-BDD) products in current clinical use and in the drug development pipeline, as well as in experimental FVIII gene therapy strategies. Here, we report that processing by furin is in fact deleterious to FVIII-BDD secretion and procoagulant activity. Inhibition of furin increases the secretion and decreases the intracellular retention of FVIII-BDD protein in mammalian cells. Our new variant (FVIII-ΔF), in which this recognition motif is removed, efficiently circumvents furin. FVIII-ΔF demonstrates increased recombinant protein yields, enhanced clotting activity, and higher circulating FVIII levels after adeno-associated viral vector-based liver gene therapy in a murine model of severe hemophilia A (HA) compared with FVIII-BDD. Moreover, we observed an amelioration of the bleeding phenotype in severe HA dogs with sustained therapeutic FVIII levels after FVIII-ΔF gene therapy at a lower vector dose than previously employed in this model. The immunogenicity of FVIII-ΔF did not differ from that of FVIII-BDD as a protein or a gene therapeutic. Thus, contrary to previous suppositions, FVIII variants that can avoid furin processing are likely to have enhanced translational potential for HA therapy.
doi_str_mv 10.1172/jci.insight.89371
format Article
fullrecord <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5053153</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1835403215</sourcerecordid><originalsourceid>FETCH-LOGICAL-c469t-9b474b9b260e7aa439666c07f857310259c7c714a5fb2de87bd6d075188183a53</originalsourceid><addsrcrecordid>eNpVkcFu3CAQhlHVqom2eYBeKo657AaMMfYlUrRq2pUi9dL2ijAeryfC4IC90r592GYbJSfQMPP9wEfIV842nKvi5tHiBn3C_TBv6kYo_oFcFkI1a6FY_fHN_oJcpfTIGOOqLJisP5OLQilRMlFekmWL0S7jAfyMfk_7JaKn4AfjLSTaGzuHSP_udjvaYnBhj9Y4mqt4wPlIje-oGcFhiGbO_a0D6E6caQAf5uOUa5k3wBimAR0aOoYOXPpCPvXGJbg6ryvy5_777-3P9cOvH7vt3cPallUzr5u2VGXbtEXFQBlTiqaqKstUX0slOCtkY5VVvDSyb4sOatV2VceU5HXNa2GkWJHbF-60tCN0Nr8yGqeniKOJRx0M6vcnHge9DwctmRRcigy4PgNieFogzXrEZME54yEsSecYmT-y4Kcs_tJqY0gpQv8aw5k-GdPZmD4b0_-M5Zlvb-_3OvHfj3gG6_WXmQ</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1835403215</pqid></control><display><type>article</type><title>Circumventing furin enhances factor VIII biological activity and ameliorates bleeding phenotypes in hemophilia models</title><source>PubMed (Medline)</source><source>MEDLINE</source><source>EZB Electronic Journals Library</source><creator>Siner, Joshua I ; Samelson-Jones, Benjamin J ; Crudele, Julie M ; French, Robert A ; Lee, Benjamin J ; Zhou, Shanzhen ; Merricks, Elizabeth ; Raymer, Robin ; Nichols, Timothy C ; Camire, Rodney M ; Arruda, Valder R</creator><creatorcontrib>Siner, Joshua I ; Samelson-Jones, Benjamin J ; Crudele, Julie M ; French, Robert A ; Lee, Benjamin J ; Zhou, Shanzhen ; Merricks, Elizabeth ; Raymer, Robin ; Nichols, Timothy C ; Camire, Rodney M ; Arruda, Valder R</creatorcontrib><description>Processing by the proprotein convertase furin is believed to be critical for the biological activity of multiple proteins involved in hemostasis, including coagulation factor VIII (FVIII). This belief prompted the retention of the furin recognition motif (amino acids 1645-1648) in the design of B-domain-deleted FVIII (FVIII-BDD) products in current clinical use and in the drug development pipeline, as well as in experimental FVIII gene therapy strategies. Here, we report that processing by furin is in fact deleterious to FVIII-BDD secretion and procoagulant activity. Inhibition of furin increases the secretion and decreases the intracellular retention of FVIII-BDD protein in mammalian cells. Our new variant (FVIII-ΔF), in which this recognition motif is removed, efficiently circumvents furin. FVIII-ΔF demonstrates increased recombinant protein yields, enhanced clotting activity, and higher circulating FVIII levels after adeno-associated viral vector-based liver gene therapy in a murine model of severe hemophilia A (HA) compared with FVIII-BDD. Moreover, we observed an amelioration of the bleeding phenotype in severe HA dogs with sustained therapeutic FVIII levels after FVIII-ΔF gene therapy at a lower vector dose than previously employed in this model. The immunogenicity of FVIII-ΔF did not differ from that of FVIII-BDD as a protein or a gene therapeutic. Thus, contrary to previous suppositions, FVIII variants that can avoid furin processing are likely to have enhanced translational potential for HA therapy.</description><identifier>ISSN: 2379-3708</identifier><identifier>EISSN: 2379-3708</identifier><identifier>DOI: 10.1172/jci.insight.89371</identifier><identifier>PMID: 27734034</identifier><language>eng</language><publisher>United States: American Society for Clinical Investigation</publisher><subject>Animals ; Cell Line ; Dogs ; Factor VIII - genetics ; Factor VIII - therapeutic use ; Furin - metabolism ; Genetic Therapy ; Hemophilia A - genetics ; Hemophilia A - therapy ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Phenotype ; Recombinant Proteins - therapeutic use</subject><ispartof>JCI insight, 2016-10, Vol.1 (16), p.e89371-e89371</ispartof><rights>Copyright © 2016, American Society for Clinical Investigation 2016 American Society for Clinical Investigation</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c469t-9b474b9b260e7aa439666c07f857310259c7c714a5fb2de87bd6d075188183a53</citedby><cites>FETCH-LOGICAL-c469t-9b474b9b260e7aa439666c07f857310259c7c714a5fb2de87bd6d075188183a53</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/PMC5053153/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5053153/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27734034$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Siner, Joshua I</creatorcontrib><creatorcontrib>Samelson-Jones, Benjamin J</creatorcontrib><creatorcontrib>Crudele, Julie M</creatorcontrib><creatorcontrib>French, Robert A</creatorcontrib><creatorcontrib>Lee, Benjamin J</creatorcontrib><creatorcontrib>Zhou, Shanzhen</creatorcontrib><creatorcontrib>Merricks, Elizabeth</creatorcontrib><creatorcontrib>Raymer, Robin</creatorcontrib><creatorcontrib>Nichols, Timothy C</creatorcontrib><creatorcontrib>Camire, Rodney M</creatorcontrib><creatorcontrib>Arruda, Valder R</creatorcontrib><title>Circumventing furin enhances factor VIII biological activity and ameliorates bleeding phenotypes in hemophilia models</title><title>JCI insight</title><addtitle>JCI Insight</addtitle><description>Processing by the proprotein convertase furin is believed to be critical for the biological activity of multiple proteins involved in hemostasis, including coagulation factor VIII (FVIII). This belief prompted the retention of the furin recognition motif (amino acids 1645-1648) in the design of B-domain-deleted FVIII (FVIII-BDD) products in current clinical use and in the drug development pipeline, as well as in experimental FVIII gene therapy strategies. Here, we report that processing by furin is in fact deleterious to FVIII-BDD secretion and procoagulant activity. Inhibition of furin increases the secretion and decreases the intracellular retention of FVIII-BDD protein in mammalian cells. Our new variant (FVIII-ΔF), in which this recognition motif is removed, efficiently circumvents furin. FVIII-ΔF demonstrates increased recombinant protein yields, enhanced clotting activity, and higher circulating FVIII levels after adeno-associated viral vector-based liver gene therapy in a murine model of severe hemophilia A (HA) compared with FVIII-BDD. Moreover, we observed an amelioration of the bleeding phenotype in severe HA dogs with sustained therapeutic FVIII levels after FVIII-ΔF gene therapy at a lower vector dose than previously employed in this model. The immunogenicity of FVIII-ΔF did not differ from that of FVIII-BDD as a protein or a gene therapeutic. Thus, contrary to previous suppositions, FVIII variants that can avoid furin processing are likely to have enhanced translational potential for HA therapy.</description><subject>Animals</subject><subject>Cell Line</subject><subject>Dogs</subject><subject>Factor VIII - genetics</subject><subject>Factor VIII - therapeutic use</subject><subject>Furin - metabolism</subject><subject>Genetic Therapy</subject><subject>Hemophilia A - genetics</subject><subject>Hemophilia A - therapy</subject><subject>Humans</subject><subject>Male</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Mice, Knockout</subject><subject>Phenotype</subject><subject>Recombinant Proteins - therapeutic use</subject><issn>2379-3708</issn><issn>2379-3708</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkcFu3CAQhlHVqom2eYBeKo657AaMMfYlUrRq2pUi9dL2ijAeryfC4IC90r592GYbJSfQMPP9wEfIV842nKvi5tHiBn3C_TBv6kYo_oFcFkI1a6FY_fHN_oJcpfTIGOOqLJisP5OLQilRMlFekmWL0S7jAfyMfk_7JaKn4AfjLSTaGzuHSP_udjvaYnBhj9Y4mqt4wPlIje-oGcFhiGbO_a0D6E6caQAf5uOUa5k3wBimAR0aOoYOXPpCPvXGJbg6ryvy5_777-3P9cOvH7vt3cPallUzr5u2VGXbtEXFQBlTiqaqKstUX0slOCtkY5VVvDSyb4sOatV2VceU5HXNa2GkWJHbF-60tCN0Nr8yGqeniKOJRx0M6vcnHge9DwctmRRcigy4PgNieFogzXrEZME54yEsSecYmT-y4Kcs_tJqY0gpQv8aw5k-GdPZmD4b0_-M5Zlvb-_3OvHfj3gG6_WXmQ</recordid><startdate>20161006</startdate><enddate>20161006</enddate><creator>Siner, Joshua I</creator><creator>Samelson-Jones, Benjamin J</creator><creator>Crudele, Julie M</creator><creator>French, Robert A</creator><creator>Lee, Benjamin J</creator><creator>Zhou, Shanzhen</creator><creator>Merricks, Elizabeth</creator><creator>Raymer, Robin</creator><creator>Nichols, Timothy C</creator><creator>Camire, Rodney M</creator><creator>Arruda, Valder R</creator><general>American Society for Clinical Investigation</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>7X8</scope><scope>5PM</scope></search><sort><creationdate>20161006</creationdate><title>Circumventing furin enhances factor VIII biological activity and ameliorates bleeding phenotypes in hemophilia models</title><author>Siner, Joshua I ; Samelson-Jones, Benjamin J ; Crudele, Julie M ; French, Robert A ; Lee, Benjamin J ; Zhou, Shanzhen ; Merricks, Elizabeth ; Raymer, Robin ; Nichols, Timothy C ; Camire, Rodney M ; Arruda, Valder R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c469t-9b474b9b260e7aa439666c07f857310259c7c714a5fb2de87bd6d075188183a53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Animals</topic><topic>Cell Line</topic><topic>Dogs</topic><topic>Factor VIII - genetics</topic><topic>Factor VIII - therapeutic use</topic><topic>Furin - metabolism</topic><topic>Genetic Therapy</topic><topic>Hemophilia A - genetics</topic><topic>Hemophilia A - therapy</topic><topic>Humans</topic><topic>Male</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Mice, Knockout</topic><topic>Phenotype</topic><topic>Recombinant Proteins - therapeutic use</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Siner, Joshua I</creatorcontrib><creatorcontrib>Samelson-Jones, Benjamin J</creatorcontrib><creatorcontrib>Crudele, Julie M</creatorcontrib><creatorcontrib>French, Robert A</creatorcontrib><creatorcontrib>Lee, Benjamin J</creatorcontrib><creatorcontrib>Zhou, Shanzhen</creatorcontrib><creatorcontrib>Merricks, Elizabeth</creatorcontrib><creatorcontrib>Raymer, Robin</creatorcontrib><creatorcontrib>Nichols, Timothy C</creatorcontrib><creatorcontrib>Camire, Rodney M</creatorcontrib><creatorcontrib>Arruda, Valder R</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>JCI insight</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Siner, Joshua I</au><au>Samelson-Jones, Benjamin J</au><au>Crudele, Julie M</au><au>French, Robert A</au><au>Lee, Benjamin J</au><au>Zhou, Shanzhen</au><au>Merricks, Elizabeth</au><au>Raymer, Robin</au><au>Nichols, Timothy C</au><au>Camire, Rodney M</au><au>Arruda, Valder R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Circumventing furin enhances factor VIII biological activity and ameliorates bleeding phenotypes in hemophilia models</atitle><jtitle>JCI insight</jtitle><addtitle>JCI Insight</addtitle><date>2016-10-06</date><risdate>2016</risdate><volume>1</volume><issue>16</issue><spage>e89371</spage><epage>e89371</epage><pages>e89371-e89371</pages><issn>2379-3708</issn><eissn>2379-3708</eissn><abstract>Processing by the proprotein convertase furin is believed to be critical for the biological activity of multiple proteins involved in hemostasis, including coagulation factor VIII (FVIII). This belief prompted the retention of the furin recognition motif (amino acids 1645-1648) in the design of B-domain-deleted FVIII (FVIII-BDD) products in current clinical use and in the drug development pipeline, as well as in experimental FVIII gene therapy strategies. Here, we report that processing by furin is in fact deleterious to FVIII-BDD secretion and procoagulant activity. Inhibition of furin increases the secretion and decreases the intracellular retention of FVIII-BDD protein in mammalian cells. Our new variant (FVIII-ΔF), in which this recognition motif is removed, efficiently circumvents furin. FVIII-ΔF demonstrates increased recombinant protein yields, enhanced clotting activity, and higher circulating FVIII levels after adeno-associated viral vector-based liver gene therapy in a murine model of severe hemophilia A (HA) compared with FVIII-BDD. Moreover, we observed an amelioration of the bleeding phenotype in severe HA dogs with sustained therapeutic FVIII levels after FVIII-ΔF gene therapy at a lower vector dose than previously employed in this model. The immunogenicity of FVIII-ΔF did not differ from that of FVIII-BDD as a protein or a gene therapeutic. Thus, contrary to previous suppositions, FVIII variants that can avoid furin processing are likely to have enhanced translational potential for HA therapy.</abstract><cop>United States</cop><pub>American Society for Clinical Investigation</pub><pmid>27734034</pmid><doi>10.1172/jci.insight.89371</doi><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 2379-3708
ispartof JCI insight, 2016-10, Vol.1 (16), p.e89371-e89371
issn 2379-3708
2379-3708
language eng
recordid cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5053153
source PubMed (Medline); MEDLINE; EZB Electronic Journals Library
subjects Animals
Cell Line
Dogs
Factor VIII - genetics
Factor VIII - therapeutic use
Furin - metabolism
Genetic Therapy
Hemophilia A - genetics
Hemophilia A - therapy
Humans
Male
Mice
Mice, Inbred C57BL
Mice, Knockout
Phenotype
Recombinant Proteins - therapeutic use
title Circumventing furin enhances factor VIII biological activity and ameliorates bleeding phenotypes in hemophilia models
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T10%3A13%3A44IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Circumventing%20furin%20enhances%20factor%20VIII%20biological%20activity%20and%20ameliorates%20bleeding%20phenotypes%20in%20hemophilia%20models&rft.jtitle=JCI%20insight&rft.au=Siner,%20Joshua%20I&rft.date=2016-10-06&rft.volume=1&rft.issue=16&rft.spage=e89371&rft.epage=e89371&rft.pages=e89371-e89371&rft.issn=2379-3708&rft.eissn=2379-3708&rft_id=info:doi/10.1172/jci.insight.89371&rft_dat=%3Cproquest_pubme%3E1835403215%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1835403215&rft_id=info:pmid/27734034&rfr_iscdi=true