Lentiviral RNAi-induced downregulation of adenosine kinase in human mesenchymal stem cell grafts: A novel perspective for seizure control
Cell therapies based on focal delivery of the inhibitory neuromodulator adenosine were previously shown to provide potent seizure suppression in animal models of epilepsy. However, hitherto used therapeutic cells were derived from rodents and thus not suitable for clinical applications. Autologous p...
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Veröffentlicht in: | Experimental neurology 2007-11, Vol.208 (1), p.26-37 |
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description | Cell therapies based on focal delivery of the inhibitory neuromodulator adenosine were previously shown to provide potent seizure suppression in animal models of epilepsy. However, hitherto used therapeutic cells were derived from rodents and thus not suitable for clinical applications. Autologous patient-derived adenosine-releasing cell implants would constitute a major therapeutic advance to avoid both xenotransplantation and immunosuppression. Here we describe a novel approach based on lentiviral RNAi mediated downregulation of adenosine kinase (ADK), the major adenosine-removing enzyme, in human mesenchymal stem cells (hMSCs), which would be compatible with autologous cell grafting in patients. Following lentiviral transduction of hMSCs with anti-ADK miRNA expression cassettes we demonstrate up to 80% downregulation of ADK and a concentration of 8.5 ng adenosine per ml of medium after incubating 10
5 cells for 8 h. hMSCs with a knockdown of ADK or cells expressing a scrambled control sequence were transplanted into hippocampi of mice 1 week prior to the intraamygdaloid injection of kainic acid (KA). While mice with control implants expressing a scrambled miRNA sequence or sham treated control animals were characterized by KA-induced status epilepticus and subsequent CA3 neuronal cell loss, animals with therapeutic ADK knockdown implants displayed a 35% reduction in seizure duration and 65% reduction in CA3 neuronal cell loss, when analyzed 24 h after KA-injection. We conclude that lentiviral expression of anti-ADK miRNA constitutes a versatile tool to generate therapeutically effective adenosine releasing hMSCs, thus representing a model system to generate patient identical autologous adult stem cell grafts. |
doi_str_mv | 10.1016/j.expneurol.2007.07.016 |
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5 cells for 8 h. hMSCs with a knockdown of ADK or cells expressing a scrambled control sequence were transplanted into hippocampi of mice 1 week prior to the intraamygdaloid injection of kainic acid (KA). While mice with control implants expressing a scrambled miRNA sequence or sham treated control animals were characterized by KA-induced status epilepticus and subsequent CA3 neuronal cell loss, animals with therapeutic ADK knockdown implants displayed a 35% reduction in seizure duration and 65% reduction in CA3 neuronal cell loss, when analyzed 24 h after KA-injection. We conclude that lentiviral expression of anti-ADK miRNA constitutes a versatile tool to generate therapeutically effective adenosine releasing hMSCs, thus representing a model system to generate patient identical autologous adult stem cell grafts.</description><identifier>ISSN: 0014-4886</identifier><identifier>EISSN: 1090-2430</identifier><identifier>DOI: 10.1016/j.expneurol.2007.07.016</identifier><identifier>PMID: 17716659</identifier><identifier>CODEN: EXNEAC</identifier><language>eng</language><publisher>Amsterdam: Elsevier Inc</publisher><subject>Adenosine ; Adenosine - metabolism ; Adenosine kinase ; Adenosine Kinase - genetics ; Adenosine Kinase - metabolism ; Animals ; Biological and medical sciences ; Cell therapy ; Down-Regulation ; Epilepsy ; Excitatory Amino Acid Agonists ; Genetic Vectors ; Headache. Facial pains. Syncopes. Epilepsia. Intracranial hypertension. Brain oedema. Cerebral palsy ; Hippocampus - pathology ; Hippocampus - physiopathology ; Human mesenchymal stem cells ; Humans ; Kainic Acid ; Lentivirus ; Lentivirus - genetics ; Male ; Medical sciences ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells - enzymology ; Mesenchymal Stromal Cells - metabolism ; Mice ; Mice, Inbred C57BL ; Nervous system (semeiology, syndromes) ; Neurology ; Neuroprotection ; RNA Interference ; RNAi ; Status epilepticus ; Status Epilepticus - chemically induced ; Status Epilepticus - pathology ; Status Epilepticus - physiopathology ; Status Epilepticus - surgery ; Transduction, Genetic</subject><ispartof>Experimental neurology, 2007-11, Vol.208 (1), p.26-37</ispartof><rights>2007 Elsevier Inc.</rights><rights>2008 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c534t-b3ce02f92d59694bca175f55713785315345dfb836417b93b289d569d8749f213</citedby><cites>FETCH-LOGICAL-c534t-b3ce02f92d59694bca175f55713785315345dfb836417b93b289d569d8749f213</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.expneurol.2007.07.016$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,780,784,885,3548,27923,27924,45994</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19873362$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17716659$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ren, Gaoying</creatorcontrib><creatorcontrib>Li, Tianfu</creatorcontrib><creatorcontrib>Lan, Jiang Quan</creatorcontrib><creatorcontrib>Wilz, Andrew</creatorcontrib><creatorcontrib>Simon, Roger P.</creatorcontrib><creatorcontrib>Boison, Detlev</creatorcontrib><title>Lentiviral RNAi-induced downregulation of adenosine kinase in human mesenchymal stem cell grafts: A novel perspective for seizure control</title><title>Experimental neurology</title><addtitle>Exp Neurol</addtitle><description>Cell therapies based on focal delivery of the inhibitory neuromodulator adenosine were previously shown to provide potent seizure suppression in animal models of epilepsy. However, hitherto used therapeutic cells were derived from rodents and thus not suitable for clinical applications. Autologous patient-derived adenosine-releasing cell implants would constitute a major therapeutic advance to avoid both xenotransplantation and immunosuppression. Here we describe a novel approach based on lentiviral RNAi mediated downregulation of adenosine kinase (ADK), the major adenosine-removing enzyme, in human mesenchymal stem cells (hMSCs), which would be compatible with autologous cell grafting in patients. Following lentiviral transduction of hMSCs with anti-ADK miRNA expression cassettes we demonstrate up to 80% downregulation of ADK and a concentration of 8.5 ng adenosine per ml of medium after incubating 10
5 cells for 8 h. hMSCs with a knockdown of ADK or cells expressing a scrambled control sequence were transplanted into hippocampi of mice 1 week prior to the intraamygdaloid injection of kainic acid (KA). While mice with control implants expressing a scrambled miRNA sequence or sham treated control animals were characterized by KA-induced status epilepticus and subsequent CA3 neuronal cell loss, animals with therapeutic ADK knockdown implants displayed a 35% reduction in seizure duration and 65% reduction in CA3 neuronal cell loss, when analyzed 24 h after KA-injection. We conclude that lentiviral expression of anti-ADK miRNA constitutes a versatile tool to generate therapeutically effective adenosine releasing hMSCs, thus representing a model system to generate patient identical autologous adult stem cell grafts.</description><subject>Adenosine</subject><subject>Adenosine - metabolism</subject><subject>Adenosine kinase</subject><subject>Adenosine Kinase - genetics</subject><subject>Adenosine Kinase - metabolism</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cell therapy</subject><subject>Down-Regulation</subject><subject>Epilepsy</subject><subject>Excitatory Amino Acid Agonists</subject><subject>Genetic Vectors</subject><subject>Headache. Facial pains. Syncopes. Epilepsia. Intracranial hypertension. Brain oedema. Cerebral palsy</subject><subject>Hippocampus - pathology</subject><subject>Hippocampus - physiopathology</subject><subject>Human mesenchymal stem cells</subject><subject>Humans</subject><subject>Kainic Acid</subject><subject>Lentivirus</subject><subject>Lentivirus - genetics</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Mesenchymal Stem Cell Transplantation</subject><subject>Mesenchymal Stromal Cells - enzymology</subject><subject>Mesenchymal Stromal Cells - metabolism</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Nervous system (semeiology, syndromes)</subject><subject>Neurology</subject><subject>Neuroprotection</subject><subject>RNA Interference</subject><subject>RNAi</subject><subject>Status epilepticus</subject><subject>Status Epilepticus - chemically induced</subject><subject>Status Epilepticus - pathology</subject><subject>Status Epilepticus - physiopathology</subject><subject>Status Epilepticus - surgery</subject><subject>Transduction, Genetic</subject><issn>0014-4886</issn><issn>1090-2430</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcGO0zAQhiMEYsvCK4AvcEuxndiOOSBVKxaQKpAQnC3HnrQuiR3spLD7Brw1jlrtwglpJB_8zT_zz18ULwheE0z468Mafo0e5hj6NcVYrJci_EGxIljiktYVflisMCZ1WTcNvyiepHTAGMuaisfFBRGCcM7kqvi9BT-5o4u6R18-bVzpvJ0NWGTDTx9hN_d6csGj0CFtwYfkPKDvzusEyHm0nwft0QAJvNnfDFkkTTAgA32PdlF3U3qDNsiHI_RohJhGMHkaoC5ElMDdzhGQCX7KPp4WjzrdJ3h2fi-Lb9fvvl59KLef33-82mxLw6p6KtvKAKadpJZJLuvWaCJYx5gglWhYRTLEbNc2Fa-JaGXV0kZaxqVtRC07SqrL4u1Jd5zbAazJ_rN5NUY36Hijgnbq3x_v9moXjopSzBhtssCrs0AMP2ZIkxpcWhxrD2FOimLKG0lZBsUJNDGkFKG7G0KwWmJUB3UXo1piVEsRnjuf_73jfd85twy8PAM6Gd13UXvj0j0nG1FVnGZuc-IgX_ToIKpkXM4KrIs5CmWD--8yfwDiqMOn</recordid><startdate>20071101</startdate><enddate>20071101</enddate><creator>Ren, Gaoying</creator><creator>Li, Tianfu</creator><creator>Lan, Jiang Quan</creator><creator>Wilz, Andrew</creator><creator>Simon, Roger P.</creator><creator>Boison, Detlev</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>IQODW</scope><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>7QO</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>5PM</scope></search><sort><creationdate>20071101</creationdate><title>Lentiviral RNAi-induced downregulation of adenosine kinase in human mesenchymal stem cell grafts: A novel perspective for seizure control</title><author>Ren, Gaoying ; Li, Tianfu ; Lan, Jiang Quan ; Wilz, Andrew ; Simon, Roger P. ; Boison, Detlev</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c534t-b3ce02f92d59694bca175f55713785315345dfb836417b93b289d569d8749f213</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Adenosine</topic><topic>Adenosine - metabolism</topic><topic>Adenosine kinase</topic><topic>Adenosine Kinase - genetics</topic><topic>Adenosine Kinase - metabolism</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Cell therapy</topic><topic>Down-Regulation</topic><topic>Epilepsy</topic><topic>Excitatory Amino Acid Agonists</topic><topic>Genetic Vectors</topic><topic>Headache. Facial pains. Syncopes. Epilepsia. Intracranial hypertension. Brain oedema. Cerebral palsy</topic><topic>Hippocampus - pathology</topic><topic>Hippocampus - physiopathology</topic><topic>Human mesenchymal stem cells</topic><topic>Humans</topic><topic>Kainic Acid</topic><topic>Lentivirus</topic><topic>Lentivirus - genetics</topic><topic>Male</topic><topic>Medical sciences</topic><topic>Mesenchymal Stem Cell Transplantation</topic><topic>Mesenchymal Stromal Cells - enzymology</topic><topic>Mesenchymal Stromal Cells - metabolism</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Nervous system (semeiology, syndromes)</topic><topic>Neurology</topic><topic>Neuroprotection</topic><topic>RNA Interference</topic><topic>RNAi</topic><topic>Status epilepticus</topic><topic>Status Epilepticus - chemically induced</topic><topic>Status Epilepticus - pathology</topic><topic>Status Epilepticus - physiopathology</topic><topic>Status Epilepticus - surgery</topic><topic>Transduction, Genetic</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ren, Gaoying</creatorcontrib><creatorcontrib>Li, Tianfu</creatorcontrib><creatorcontrib>Lan, Jiang Quan</creatorcontrib><creatorcontrib>Wilz, Andrew</creatorcontrib><creatorcontrib>Simon, Roger P.</creatorcontrib><creatorcontrib>Boison, Detlev</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Experimental neurology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ren, Gaoying</au><au>Li, Tianfu</au><au>Lan, Jiang Quan</au><au>Wilz, Andrew</au><au>Simon, Roger P.</au><au>Boison, Detlev</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Lentiviral RNAi-induced downregulation of adenosine kinase in human mesenchymal stem cell grafts: A novel perspective for seizure control</atitle><jtitle>Experimental neurology</jtitle><addtitle>Exp Neurol</addtitle><date>2007-11-01</date><risdate>2007</risdate><volume>208</volume><issue>1</issue><spage>26</spage><epage>37</epage><pages>26-37</pages><issn>0014-4886</issn><eissn>1090-2430</eissn><coden>EXNEAC</coden><abstract>Cell therapies based on focal delivery of the inhibitory neuromodulator adenosine were previously shown to provide potent seizure suppression in animal models of epilepsy. However, hitherto used therapeutic cells were derived from rodents and thus not suitable for clinical applications. Autologous patient-derived adenosine-releasing cell implants would constitute a major therapeutic advance to avoid both xenotransplantation and immunosuppression. Here we describe a novel approach based on lentiviral RNAi mediated downregulation of adenosine kinase (ADK), the major adenosine-removing enzyme, in human mesenchymal stem cells (hMSCs), which would be compatible with autologous cell grafting in patients. Following lentiviral transduction of hMSCs with anti-ADK miRNA expression cassettes we demonstrate up to 80% downregulation of ADK and a concentration of 8.5 ng adenosine per ml of medium after incubating 10
5 cells for 8 h. hMSCs with a knockdown of ADK or cells expressing a scrambled control sequence were transplanted into hippocampi of mice 1 week prior to the intraamygdaloid injection of kainic acid (KA). While mice with control implants expressing a scrambled miRNA sequence or sham treated control animals were characterized by KA-induced status epilepticus and subsequent CA3 neuronal cell loss, animals with therapeutic ADK knockdown implants displayed a 35% reduction in seizure duration and 65% reduction in CA3 neuronal cell loss, when analyzed 24 h after KA-injection. We conclude that lentiviral expression of anti-ADK miRNA constitutes a versatile tool to generate therapeutically effective adenosine releasing hMSCs, thus representing a model system to generate patient identical autologous adult stem cell grafts.</abstract><cop>Amsterdam</cop><pub>Elsevier Inc</pub><pmid>17716659</pmid><doi>10.1016/j.expneurol.2007.07.016</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Adenosine Adenosine - metabolism Adenosine kinase Adenosine Kinase - genetics Adenosine Kinase - metabolism Animals Biological and medical sciences Cell therapy Down-Regulation Epilepsy Excitatory Amino Acid Agonists Genetic Vectors Headache. Facial pains. Syncopes. Epilepsia. Intracranial hypertension. Brain oedema. Cerebral palsy Hippocampus - pathology Hippocampus - physiopathology Human mesenchymal stem cells Humans Kainic Acid Lentivirus Lentivirus - genetics Male Medical sciences Mesenchymal Stem Cell Transplantation Mesenchymal Stromal Cells - enzymology Mesenchymal Stromal Cells - metabolism Mice Mice, Inbred C57BL Nervous system (semeiology, syndromes) Neurology Neuroprotection RNA Interference RNAi Status epilepticus Status Epilepticus - chemically induced Status Epilepticus - pathology Status Epilepticus - physiopathology Status Epilepticus - surgery Transduction, Genetic |
title | Lentiviral RNAi-induced downregulation of adenosine kinase in human mesenchymal stem cell grafts: A novel perspective for seizure control |
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