Efficient Generation of A9 Midbrain Dopaminergic Neurons by Lentiviral Delivery of LMX1A in Human Embryonic Stem Cells and Induced Pluripotent Stem Cells
Identifieur interne : 000A87 ( PascalFrancis/Curation ); précédent : 000A86; suivant : 000A88Efficient Generation of A9 Midbrain Dopaminergic Neurons by Lentiviral Delivery of LMX1A in Human Embryonic Stem Cells and Induced Pluripotent Stem Cells
Auteurs : A. Sanchez-Danes [Espagne] ; A. Consiglio [Espagne, Italie] ; Y. Richaud [Espagne] ; I. Rodriguez-Piza [Espagne] ; B. Dehay [Espagne, France] ; M. Edel [Espagne, Australie] ; J. Bove [Espagne] ; M. Memo [Italie] ; M. Vila [Espagne] ; A. Raya [Espagne] ; J. C. Izpisua Belmonte [Espagne, Canada]Source :
- Human gene therapy [ 1043-0342 ] ; 2012.
Descripteurs français
- Pascal (Inist)
- Wicri :
- topic : Homme, Cellule souche.
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Abstract
Human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) offer great hope for in vitro modeling of Parkinson's disease (PD), as well as for designing cell-replacement therapies. To realize these opportunities, there is an urgent need to develop efficient protocols for the directed differentiation of hESC/iPSC into dopamine (DA) neurons with the specific characteristics of the cell population lost to PD, i.e., A9-subtype ventral midbrain DA neurons. Here we use lentiviral vectors to drive the expression of LMX1A, which encodes a transcription factor critical for ventral midbrain identity, specifically in neural progenitor cells. We show that clonal lines of hESC engineered to contain one or two copies of this lentiviral vector retain long-term self-renewing ability and pluripotent differentiation capacity. Greater than 60% of all neurons generated from LMX1A-engineered hESC were ventral midbrain DA neurons of the A9 subtype, compared with ∼10% in green fluorescent protein-engineered controls, as judged by specific marker expression and functional analyses. Moreover, DA neuron precursors differentiated from LMX1A-engineered hESC were able to survive and differentiate when grafted into the brain of adult mice. Finally, we provide evidence that LMX1A overexpression similarly increases the yield of DA neuron differentiation from human iPSC. Taken together, our data show that stable genetic engineering of hESC/iPSC with lentiviral vectors driving controlled expression of LMX1A is an efficient way to generate enriched populations of human A9-subtype ventral midbrain DA neurons, which should prove useful for modeling PD and may be helpful for designing future cell-replacement strategies.
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en" level="a">Efficient Generation of A9 Midbrain Dopaminergic Neurons by Lentiviral Delivery of LMX1A in Human Embryonic Stem Cells and Induced Pluripotent Stem Cells</title><author><name sortKey="Sanchez Danes, A" sort="Sanchez Danes, A" uniqKey="Sanchez Danes A" first="A." last="Sanchez-Danes">A. Sanchez-Danes</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Biomedicine of the University of Barcelona (IBUB)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Center for Regenerative Medicine in Barcelona</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Consiglio, A" sort="Consiglio, A" uniqKey="Consiglio A" first="A." last="Consiglio">A. Consiglio</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Biomedicine of the University of Barcelona (IBUB)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Center for Regenerative Medicine in Barcelona</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Department of Biomedical Science and Biotechnology, University of Brescia</s1><s2>Brescia</s2><s3>ITA</s3><sZ>2 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Italie</country></affiliation></author><author><name sortKey="Richaud, Y" sort="Richaud, Y" uniqKey="Richaud Y" first="Y." last="Richaud">Y. Richaud</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Control of Stem Cell Potency Group, Institute for Bioengineering of Catalonia (IBEC)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>7 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Networking Center of Biomedical Research in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Rodriguez Piza, I" sort="Rodriguez Piza, I" uniqKey="Rodriguez Piza I" first="I." last="Rodriguez-Piza">I. Rodriguez-Piza</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Center for Regenerative Medicine in Barcelona</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Dehay, B" sort="Dehay, B" uniqKey="Dehay B" first="B." last="Dehay">B. Dehay</name><affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Neurodegenerative Diseases Research Group, Vall d'Hebron Research Institute</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="07"><s1>Networking Center of Biomedical Research on Neurodegenerative Diseases (CIBERNED)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="08"><s1>Institut des Maladies Neurodégénératives, Université de Bordeaux and CNRS, UMR 5293</s1><s2>33000 Bordeaux</s2><s3>FRA</s3><sZ>5 aut.</sZ></inist:fA14><country>France</country></affiliation></author><author><name sortKey="Edel, M" sort="Edel, M" uniqKey="Edel M" first="M." last="Edel">M. Edel</name><affiliation wicri:level="1"><inist:fA14 i1="09"><s1>Research Institute of Hospital Vall d'Hebron and Banc de Sang i Teixits, Advanced Cell Therapies and Immunology</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>6 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="12"><s1>Visiting Research Fellow at the Victor Chang Cardiac Research Institute</s1><s2>Sydney</s2><s3>AUS</s3><sZ>6 aut.</sZ></inist:fA14><country>Australie</country></affiliation></author><author><name sortKey="Bove, J" sort="Bove, J" uniqKey="Bove J" first="J." last="Bove">J. Bove</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Control of Stem Cell Potency Group, Institute for Bioengineering of Catalonia (IBEC)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>7 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Neurodegenerative Diseases Research Group, Vall d'Hebron Research Institute</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Memo, M" sort="Memo, M" uniqKey="Memo M" first="M." last="Memo">M. Memo</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Department of Biomedical Science and Biotechnology, University of Brescia</s1><s2>Brescia</s2><s3>ITA</s3><sZ>2 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Italie</country></affiliation></author><author><name sortKey="Vila, M" sort="Vila, M" uniqKey="Vila M" first="M." last="Vila">M. Vila</name><affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Neurodegenerative Diseases Research Group, Vall d'Hebron Research Institute</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="07"><s1>Networking Center of Biomedical Research on Neurodegenerative Diseases (CIBERNED)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="10"><s1>Catalan Institution for Research and Advanced Studies (ICREA)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Raya, A" sort="Raya, A" uniqKey="Raya A" first="A." last="Raya">A. Raya</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Control of Stem Cell Potency Group, Institute for Bioengineering of Catalonia (IBEC)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>7 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Networking Center of Biomedical Research in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="10"><s1>Catalan Institution for Research and Advanced Studies (ICREA)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Izpisua Belmonte, J C" sort="Izpisua Belmonte, J C" uniqKey="Izpisua Belmonte J" first="J. C." last="Izpisua Belmonte">J. C. Izpisua Belmonte</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Center for Regenerative Medicine in Barcelona</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="11"><s1>Gene Expression Laboratory, Salk Institute for Biological Studies</s1><s2>La Jolla</s2><s3>CAN</s3><sZ>11 aut.</sZ></inist:fA14><country>Canada</country></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">12-0391927</idno><date when="2012">2012</date><idno type="stanalyst">PASCAL 12-0391927 INIST</idno><idno type="RBID">Pascal:12-0391927</idno><idno type="wicri:Area/PascalFrancis/Corpus">000165</idno><idno type="wicri:Area/PascalFrancis/Curation">000A87</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Efficient Generation of A9 Midbrain Dopaminergic Neurons by Lentiviral Delivery of LMX1A in Human Embryonic Stem Cells and Induced Pluripotent Stem Cells</title><author><name sortKey="Sanchez Danes, A" sort="Sanchez Danes, A" uniqKey="Sanchez Danes A" first="A." last="Sanchez-Danes">A. Sanchez-Danes</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Biomedicine of the University of Barcelona (IBUB)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Center for Regenerative Medicine in Barcelona</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Consiglio, A" sort="Consiglio, A" uniqKey="Consiglio A" first="A." last="Consiglio">A. Consiglio</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Biomedicine of the University of Barcelona (IBUB)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Center for Regenerative Medicine in Barcelona</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Department of Biomedical Science and Biotechnology, University of Brescia</s1><s2>Brescia</s2><s3>ITA</s3><sZ>2 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Italie</country></affiliation></author><author><name sortKey="Richaud, Y" sort="Richaud, Y" uniqKey="Richaud Y" first="Y." last="Richaud">Y. Richaud</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Control of Stem Cell Potency Group, Institute for Bioengineering of Catalonia (IBEC)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>7 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Networking Center of Biomedical Research in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Rodriguez Piza, I" sort="Rodriguez Piza, I" uniqKey="Rodriguez Piza I" first="I." last="Rodriguez-Piza">I. Rodriguez-Piza</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Center for Regenerative Medicine in Barcelona</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Dehay, B" sort="Dehay, B" uniqKey="Dehay B" first="B." last="Dehay">B. Dehay</name><affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Neurodegenerative Diseases Research Group, Vall d'Hebron Research Institute</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="07"><s1>Networking Center of Biomedical Research on Neurodegenerative Diseases (CIBERNED)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="08"><s1>Institut des Maladies Neurodégénératives, Université de Bordeaux and CNRS, UMR 5293</s1><s2>33000 Bordeaux</s2><s3>FRA</s3><sZ>5 aut.</sZ></inist:fA14><country>France</country></affiliation></author><author><name sortKey="Edel, M" sort="Edel, M" uniqKey="Edel M" first="M." last="Edel">M. Edel</name><affiliation wicri:level="1"><inist:fA14 i1="09"><s1>Research Institute of Hospital Vall d'Hebron and Banc de Sang i Teixits, Advanced Cell Therapies and Immunology</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>6 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="12"><s1>Visiting Research Fellow at the Victor Chang Cardiac Research Institute</s1><s2>Sydney</s2><s3>AUS</s3><sZ>6 aut.</sZ></inist:fA14><country>Australie</country></affiliation></author><author><name sortKey="Bove, J" sort="Bove, J" uniqKey="Bove J" first="J." last="Bove">J. Bove</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Control of Stem Cell Potency Group, Institute for Bioengineering of Catalonia (IBEC)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>7 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Neurodegenerative Diseases Research Group, Vall d'Hebron Research Institute</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Memo, M" sort="Memo, M" uniqKey="Memo M" first="M." last="Memo">M. Memo</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Department of Biomedical Science and Biotechnology, University of Brescia</s1><s2>Brescia</s2><s3>ITA</s3><sZ>2 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Italie</country></affiliation></author><author><name sortKey="Vila, M" sort="Vila, M" uniqKey="Vila M" first="M." last="Vila">M. Vila</name><affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Neurodegenerative Diseases Research Group, Vall d'Hebron Research Institute</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="07"><s1>Networking Center of Biomedical Research on Neurodegenerative Diseases (CIBERNED)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="10"><s1>Catalan Institution for Research and Advanced Studies (ICREA)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Raya, A" sort="Raya, A" uniqKey="Raya A" first="A." last="Raya">A. Raya</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Control of Stem Cell Potency Group, Institute for Bioengineering of Catalonia (IBEC)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>7 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Networking Center of Biomedical Research in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="10"><s1>Catalan Institution for Research and Advanced Studies (ICREA)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country></affiliation></author><author><name sortKey="Izpisua Belmonte, J C" sort="Izpisua Belmonte, J C" uniqKey="Izpisua Belmonte J" first="J. C." last="Izpisua Belmonte">J. C. Izpisua Belmonte</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Center for Regenerative Medicine in Barcelona</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>11 aut.</sZ></inist:fA14><country>Espagne</country></affiliation><affiliation wicri:level="1"><inist:fA14 i1="11"><s1>Gene Expression Laboratory, Salk Institute for Biological Studies</s1><s2>La Jolla</s2><s3>CAN</s3><sZ>11 aut.</sZ></inist:fA14><country>Canada</country></affiliation></author></analytic><series><title level="j" type="main">Human gene therapy</title><title level="j" type="abbreviated">Hum. gene ther.</title><idno type="ISSN">1043-0342</idno><imprint><date when="2012">2012</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Human gene therapy</title><title level="j" type="abbreviated">Hum. gene ther.</title><idno type="ISSN">1043-0342</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Embryonic cell</term><term>Gene therapy</term><term>Human</term><term>Induced pluripotent stem cell</term><term>Lentivirus</term><term>Midbrain</term><term>Neuron</term><term>Stem cell</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Mésencéphale</term><term>Neurone</term><term>Homme</term><term>Cellule embryonnaire</term><term>Cellule souche</term><term>Thérapie génique</term><term>Lentivirus</term><term>Cellule souche pluripotente induite</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Homme</term><term>Cellule souche</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) offer great hope for in vitro modeling of Parkinson's disease (PD), as well as for designing cell-replacement therapies. To realize these opportunities, there is an urgent need to develop efficient protocols for the directed differentiation of hESC/iPSC into dopamine (DA) neurons with the specific characteristics of the cell population lost to PD, i.e., A9-subtype ventral midbrain DA neurons. Here we use lentiviral vectors to drive the expression of LMX1A, which encodes a transcription factor critical for ventral midbrain identity, specifically in neural progenitor cells. We show that clonal lines of hESC engineered to contain one or two copies of this lentiviral vector retain long-term self-renewing ability and pluripotent differentiation capacity. Greater than 60% of all neurons generated from LMX1A-engineered hESC were ventral midbrain DA neurons of the A9 subtype, compared with ∼10% in green fluorescent protein-engineered controls, as judged by specific marker expression and functional analyses. Moreover, DA neuron precursors differentiated from LMX1A-engineered hESC were able to survive and differentiate when grafted into the brain of adult mice. Finally, we provide evidence that LMX1A overexpression similarly increases the yield of DA neuron differentiation from human iPSC. Taken together, our data show that stable genetic engineering of hESC/iPSC with lentiviral vectors driving controlled expression of LMX1A is an efficient way to generate enriched populations of human A9-subtype ventral midbrain DA neurons, which should prove useful for modeling PD and may be helpful for designing future cell-replacement strategies.</div></front></TEI><inist><standard h6="B"><pA><fA01 i1="01" i2="1"><s0>1043-0342</s0></fA01><fA02 i1="01"><s0>HGTHE3</s0></fA02><fA03 i2="1"><s0>Hum. gene ther.</s0></fA03><fA05><s2>23</s2></fA05><fA06><s2>1</s2></fA06><fA08 i1="01" i2="1" l="ENG"><s1>Efficient Generation of A9 Midbrain Dopaminergic Neurons by Lentiviral Delivery of LMX1A in Human Embryonic Stem Cells and Induced Pluripotent Stem Cells</s1></fA08><fA11 i1="01" i2="1"><s1>SANCHEZ-DANES (A.)</s1></fA11><fA11 i1="02" i2="1"><s1>CONSIGLIO (A.)</s1></fA11><fA11 i1="03" i2="1"><s1>RICHAUD (Y.)</s1></fA11><fA11 i1="04" i2="1"><s1>RODRIGUEZ-PIZA (I.)</s1></fA11><fA11 i1="05" i2="1"><s1>DEHAY (B.)</s1></fA11><fA11 i1="06" i2="1"><s1>EDEL (M.)</s1></fA11><fA11 i1="07" i2="1"><s1>BOVE (J.)</s1></fA11><fA11 i1="08" i2="1"><s1>MEMO (M.)</s1></fA11><fA11 i1="09" i2="1"><s1>VILA (M.)</s1></fA11><fA11 i1="10" i2="1"><s1>RAYA (A.)</s1></fA11><fA11 i1="11" i2="1"><s1>IZPISUA BELMONTE (J. C.)</s1></fA11><fA14 i1="01"><s1>Institute of Biomedicine of the University of Barcelona (IBUB)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ></fA14><fA14 i1="02"><s1>Center for Regenerative Medicine in Barcelona</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>11 aut.</sZ></fA14><fA14 i1="03"><s1>Department of Biomedical Science and Biotechnology, University of Brescia</s1><s2>Brescia</s2><s3>ITA</s3><sZ>2 aut.</sZ><sZ>8 aut.</sZ></fA14><fA14 i1="04"><s1>Control of Stem Cell Potency Group, Institute for Bioengineering of Catalonia (IBEC)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>7 aut.</sZ><sZ>10 aut.</sZ></fA14><fA14 i1="05"><s1>Networking Center of Biomedical Research in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>3 aut.</sZ><sZ>10 aut.</sZ></fA14><fA14 i1="06"><s1>Neurodegenerative Diseases Research Group, Vall d'Hebron Research Institute</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></fA14><fA14 i1="07"><s1>Networking Center of Biomedical Research on Neurodegenerative Diseases (CIBERNED)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>5 aut.</sZ><sZ>9 aut.</sZ></fA14><fA14 i1="08"><s1>Institut des Maladies Neurodégénératives, Université de Bordeaux and CNRS, UMR 5293</s1><s2>33000 Bordeaux</s2><s3>FRA</s3><sZ>5 aut.</sZ></fA14><fA14 i1="09"><s1>Research Institute of Hospital Vall d'Hebron and Banc de Sang i Teixits, Advanced Cell Therapies and Immunology</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>6 aut.</sZ></fA14><fA14 i1="10"><s1>Catalan Institution for Research and Advanced Studies (ICREA)</s1><s2>Barcelona</s2><s3>ESP</s3><sZ>9 aut.</sZ><sZ>10 aut.</sZ></fA14><fA14 i1="11"><s1>Gene Expression Laboratory, Salk Institute for Biological Studies</s1><s2>La Jolla</s2><s3>CAN</s3><sZ>11 aut.</sZ></fA14><fA14 i1="12"><s1>Visiting Research Fellow at the Victor Chang Cardiac Research Institute</s1><s2>Sydney</s2><s3>AUS</s3><sZ>6 aut.</sZ></fA14><fA20><s1>56-69</s1></fA20><fA21><s1>2012</s1></fA21><fA23 i1="01"><s0>ENG</s0></fA23><fA43 i1="01"><s1>INIST</s1><s2>22641</s2><s5>354000505105110070</s5></fA43><fA44><s0>0000</s0><s1>© 2012 INIST-CNRS. All rights reserved.</s1></fA44><fA45><s0>1 p.</s0></fA45><fA47 i1="01" i2="1"><s0>12-0391927</s0></fA47><fA60><s1>P</s1></fA60><fA61><s0>A</s0></fA61><fA64 i1="01" i2="1"><s0>Human gene therapy</s0></fA64><fA66 i1="01"><s0>USA</s0></fA66><fC01 i1="01" l="ENG"><s0>Human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) offer great hope for in vitro modeling of Parkinson's disease (PD), as well as for designing cell-replacement therapies. To realize these opportunities, there is an urgent need to develop efficient protocols for the directed differentiation of hESC/iPSC into dopamine (DA) neurons with the specific characteristics of the cell population lost to PD, i.e., A9-subtype ventral midbrain DA neurons. Here we use lentiviral vectors to drive the expression of LMX1A, which encodes a transcription factor critical for ventral midbrain identity, specifically in neural progenitor cells. We show that clonal lines of hESC engineered to contain one or two copies of this lentiviral vector retain long-term self-renewing ability and pluripotent differentiation capacity. Greater than 60% of all neurons generated from LMX1A-engineered hESC were ventral midbrain DA neurons of the A9 subtype, compared with ∼10% in green fluorescent protein-engineered controls, as judged by specific marker expression and functional analyses. Moreover, DA neuron precursors differentiated from LMX1A-engineered hESC were able to survive and differentiate when grafted into the brain of adult mice. Finally, we provide evidence that LMX1A overexpression similarly increases the yield of DA neuron differentiation from human iPSC. Taken together, our data show that stable genetic engineering of hESC/iPSC with lentiviral vectors driving controlled expression of LMX1A is an efficient way to generate enriched populations of human A9-subtype ventral midbrain DA neurons, which should prove useful for modeling PD and may be helpful for designing future cell-replacement strategies.</s0></fC01><fC02 i1="01" i2="X"><s0>002A31D01D</s0></fC02><fC02 i1="02" i2="X"><s0>002B27D03</s0></fC02><fC02 i1="03" i2="X"><s0>215</s0></fC02><fC03 i1="01" i2="X" l="FRE"><s0>Mésencéphale</s0><s5>01</s5></fC03><fC03 i1="01" i2="X" l="ENG"><s0>Midbrain</s0><s5>01</s5></fC03><fC03 i1="01" i2="X" l="SPA"><s0>Mesencéfalo</s0><s5>01</s5></fC03><fC03 i1="02" i2="X" l="FRE"><s0>Neurone</s0><s5>02</s5></fC03><fC03 i1="02" i2="X" l="ENG"><s0>Neuron</s0><s5>02</s5></fC03><fC03 i1="02" i2="X" l="SPA"><s0>Neurona</s0><s5>02</s5></fC03><fC03 i1="03" i2="X" l="FRE"><s0>Homme</s0><s5>03</s5></fC03><fC03 i1="03" i2="X" l="ENG"><s0>Human</s0><s5>03</s5></fC03><fC03 i1="03" i2="X" l="SPA"><s0>Hombre</s0><s5>03</s5></fC03><fC03 i1="04" i2="X" l="FRE"><s0>Cellule embryonnaire</s0><s5>04</s5></fC03><fC03 i1="04" i2="X" l="ENG"><s0>Embryonic cell</s0><s5>04</s5></fC03><fC03 i1="04" i2="X" l="SPA"><s0>Célula embrionaria</s0><s5>04</s5></fC03><fC03 i1="05" i2="X" l="FRE"><s0>Cellule souche</s0><s5>05</s5></fC03><fC03 i1="05" i2="X" l="ENG"><s0>Stem cell</s0><s5>05</s5></fC03><fC03 i1="05" i2="X" l="SPA"><s0>Célula primitiva</s0><s5>05</s5></fC03><fC03 i1="06" i2="X" l="FRE"><s0>Thérapie génique</s0><s5>06</s5></fC03><fC03 i1="06" i2="X" l="ENG"><s0>Gene therapy</s0><s5>06</s5></fC03><fC03 i1="06" i2="X" l="SPA"><s0>Terapia génica</s0><s5>06</s5></fC03><fC03 i1="07" i2="X" l="FRE"><s0>Lentivirus</s0><s2>NW</s2><s5>16</s5></fC03><fC03 i1="07" i2="X" l="ENG"><s0>Lentivirus</s0><s2>NW</s2><s5>16</s5></fC03><fC03 i1="07" i2="X" l="SPA"><s0>Lentivirus</s0><s2>NW</s2><s5>16</s5></fC03><fC03 i1="08" i2="X" l="FRE"><s0>Cellule souche pluripotente induite</s0><s4>CD</s4><s5>96</s5></fC03><fC03 i1="08" i2="X" l="ENG"><s0>Induced pluripotent stem cell</s0><s4>CD</s4><s5>96</s5></fC03><fC07 i1="01" i2="X" l="FRE"><s0>Retroviridae</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="ENG"><s0>Retroviridae</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="SPA"><s0>Retroviridae</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="FRE"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="ENG"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="SPA"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="FRE"><s0>Système nerveux central</s0><s5>19</s5></fC07><fC07 i1="03" i2="X" l="ENG"><s0>Central nervous system</s0><s5>19</s5></fC07><fC07 i1="03" i2="X" l="SPA"><s0>Sistema nervioso central</s0><s5>19</s5></fC07><fN21><s1>303</s1></fN21><fN44 i1="01"><s1>OTO</s1></fN44><fN82><s1>OTO</s1></fN82></pA></standard></inist></record>Pour manipuler ce document sous Unix (Dilib)
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