{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T01:11:25Z","timestamp":1777943485776,"version":"3.51.4"},"reference-count":62,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"6","content-domain":{"domain":["www.ahajournals.org"],"crossmark-restriction":true},"short-container-title":["JAHA"],"published-print":{"date-parts":[[2016,6,13]]},"abstract":"\n Background<\/jats:title>\n \n Critical limb ischemia (\n CLI<\/jats:styled-content>\n ) is the extreme manifestation of peripheral artery disease, a major unmet clinical need for which lower limb amputation is the only option for many patients. After 2 decades in development, therapeutic angiogenesis has been tested clinically via intramuscular delivery of proangiogenic proteins, genes, and stem cells. Efficacy has been modest to absent, and the largest phase 3 trial of gene therapy for\n CLI<\/jats:styled-content>\n reported a worsening trend of plasmid fibroblast growth factor. In all clinical trials to date, gene therapy has used unregulated vectors with limited duration of expression. Only unregulated extended expression vectors such as adeno\u2010associated virus (\n AAV<\/jats:styled-content>\n ) and lentivirus have been tested in preclinical models.\n <\/jats:p>\n <\/jats:sec>\n \n Methods and Results<\/jats:title>\n \n We present preclinical results of ischemia (hypoxia)\u2010regulated conditionally silenced (\n CS<\/jats:styled-content>\n )\n AAV<\/jats:styled-content>\n \u2013human vascular endothelial growth factor (\n hVEGF<\/jats:styled-content>\n ) gene delivery that shows efficacy and safety in a setting where other strategies fail. In a BALB\/c mouse model of\n CLI<\/jats:styled-content>\n , we show that gene therapy with\n AAV<\/jats:styled-content>\n \u2010\n CS<\/jats:styled-content>\n \u2010\n hVEGF<\/jats:styled-content>\n , but not unregulated\n AAV<\/jats:styled-content>\n or plasmid, vectors conferred limb salvage, protection from necrosis, and vascular regeneration when delivered via intramuscular or intra\u2010arterial routes. All vector treatments conferred increased capillary density, but organized longitudinal arteries were selectively generated by\n AAV<\/jats:styled-content>\n \u2010\n CS<\/jats:styled-content>\n \u2010\n hVEGF<\/jats:styled-content>\n .\n AAV<\/jats:styled-content>\n \u2010\n CS<\/jats:styled-content>\n \u2010\n hVEGF<\/jats:styled-content>\n therapy reversibly activated angiogenic and vasculogenic genes, including\n Notch<\/jats:italic>\n ,\n \n SDF<\/jats:styled-content>\n 1\n <\/jats:italic>\n ,\n Angiopoietin<\/jats:italic>\n , and\n Ephrin\u2010B2<\/jats:italic>\n . Reoxygenation extinguished\n VEGF<\/jats:styled-content>\n expression and inactivated the program with no apparent adverse side effects.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusions<\/jats:title>\n \n Restriction of angiogenic growth factor expression to regions of ischemia supports the safe and stable reperfusion of hindlimbs in a clinically relevant murine model of\n CLI<\/jats:styled-content>\n .\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1161\/jaha.115.001815","type":"journal-article","created":{"date-parts":[[2016,5,27]],"date-time":"2016-05-27T02:58:37Z","timestamp":1464317917000},"update-policy":"https:\/\/doi.org\/10.1161\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":20,"title":["Vascular Regeneration in Ischemic Hindlimb by Adeno\u2010Associated Virus Expressing Conditionally Silenced Vascular Endothelial Growth Factor"],"prefix":"10.1161","volume":"5","author":[{"given":"Jeffrey","family":"Boden","sequence":"first","affiliation":[{"name":"Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL"},{"name":"Vascular Biology Institute, University of Miami Miller School of Medicine, Miami, FL"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roberta Marques","family":"Lassance\u2010Soares","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL"},{"name":"Vascular Biology Institute, University of Miami Miller School of Medicine, Miami, FL"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huilan","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL"},{"name":"Vascular Biology Institute, University of Miami Miller School of Medicine, Miami, FL"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuntao","family":"Wei","sequence":"additional","affiliation":[{"name":"Department of Surgery, University of Miami Miller School of Medicine, Miami, FL"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria\u2010Grazia","family":"Spiga","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jennipher","family":"Adi","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hans","family":"Layman","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, University of Miami Miller School of Medicine, Miami, FL"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hong","family":"Yu","sequence":"additional","affiliation":[{"name":"Vascular Biology Institute, University of Miami Miller School of Medicine, Miami, FL"},{"name":"Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roberto I.","family":"Vazquez\u2010Padron","sequence":"additional","affiliation":[{"name":"Department of Surgery, University of Miami Miller School of Medicine, Miami, FL"},{"name":"Vascular Biology Institute, University of Miami Miller School of Medicine, Miami, FL"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fotios","family":"Andreopoulos","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, University of Miami Miller School of Medicine, Miami, FL"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keith A.","family":"Webster","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL"},{"name":"Vascular Biology Institute, University of Miami Miller School of Medicine, Miami, FL"},{"name":"Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"276","reference":[{"key":"e_1_3_5_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jvs.2012.03.255"},{"key":"e_1_3_5_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s11886-012-0251-4"},{"key":"e_1_3_5_4_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.109.200386"},{"key":"e_1_3_5_5_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.22315"},{"key":"e_1_3_5_6_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.CIR.0000093398.16124.29"},{"key":"e_1_3_5_7_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-1241.2008.01842.x"},{"key":"e_1_3_5_8_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.107.727347"},{"key":"e_1_3_5_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(11)60394-2"},{"key":"e_1_3_5_10_2","doi-asserted-by":"publisher","DOI":"10.1253\/circj.71.196"},{"key":"e_1_3_5_11_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.110.009407"},{"key":"e_1_3_5_12_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.106.607994"},{"key":"e_1_3_5_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ahj.2007.01.038"},{"key":"e_1_3_5_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm1075"},{"key":"e_1_3_5_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2005.10.036"},{"key":"e_1_3_5_16_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.108.181115"},{"key":"e_1_3_5_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrm3176"},{"key":"e_1_3_5_18_2","doi-asserted-by":"publisher","DOI":"10.1387\/ijdb.103213sp"},{"key":"e_1_3_5_19_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI5871"},{"key":"e_1_3_5_20_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.202474099"},{"key":"e_1_3_5_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.96.24.13691"},{"key":"e_1_3_5_22_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3301514"},{"key":"e_1_3_5_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3302133"},{"key":"e_1_3_5_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2007.01.044"},{"key":"e_1_3_5_25_2","doi-asserted-by":"publisher","DOI":"10.1002\/jgm.716"},{"key":"e_1_3_5_26_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.mt.6300022"},{"key":"e_1_3_5_27_2","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2013.244"},{"key":"e_1_3_5_28_2","doi-asserted-by":"publisher","DOI":"10.2144\/000113907"},{"key":"e_1_3_5_29_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1105002108"},{"key":"e_1_3_5_30_2","doi-asserted-by":"publisher","DOI":"10.1155\/2011\/371832"},{"key":"e_1_3_5_31_2","doi-asserted-by":"publisher","DOI":"10.1152\/physiolgenomics.00047.2007"},{"key":"e_1_3_5_32_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.CIR.100.5.547"},{"key":"e_1_3_5_33_2","doi-asserted-by":"publisher","DOI":"10.2337\/diabetes.53.9.2383"},{"key":"e_1_3_5_34_2","first-page":"1315","article-title":"Activation of metallothionine gene expression by hypoxia involves metal response elements and metal transcription factor\u20101","volume":"59","author":"Murphy BJ","year":"1999","unstructured":"Murphy BJ, Andrews GK, Bittel D, Discher DJ, McCue J, Green CJ, Yanovsky M, Giaccia A, Sutherland RM, Laderoute KR, Webster KA. Activation of metallothionine gene expression by hypoxia involves metal response elements and metal transcription factor\u20101. Cancer Res. 1999;59:1315\u20131322.","journal-title":"Cancer Res"},{"key":"e_1_3_5_35_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.9.5375"},{"key":"e_1_3_5_36_2","doi-asserted-by":"publisher","DOI":"10.2174\/138161209787315783"},{"key":"e_1_3_5_37_2","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-07-6705"},{"key":"e_1_3_5_38_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.mt.6300246"},{"key":"e_1_3_5_39_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3302481"},{"key":"e_1_3_5_40_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2008.172"},{"key":"e_1_3_5_41_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jss.2009.06.021"},{"key":"e_1_3_5_42_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.291004"},{"key":"e_1_3_5_43_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature05571"},{"key":"e_1_3_5_44_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature05355"},{"key":"e_1_3_5_45_2","doi-asserted-by":"publisher","DOI":"10.2174\/138161210794455003"},{"key":"e_1_3_5_46_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2007.10.019"},{"key":"e_1_3_5_47_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature08995"},{"key":"e_1_3_5_48_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.atherosclerosis.2012.10.060"},{"key":"e_1_3_5_49_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.115.303565"},{"key":"e_1_3_5_50_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.ATV.0000202677.55012.a0"},{"key":"e_1_3_5_51_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.107.736447"},{"key":"e_1_3_5_52_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.RES.0000043281.66969.32"},{"key":"e_1_3_5_53_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2012.01.032"},{"key":"e_1_3_5_54_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.112.000860"},{"key":"e_1_3_5_55_2","doi-asserted-by":"publisher","DOI":"10.1053\/j.semvascsurg.2007.02.009"},{"key":"e_1_3_5_56_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.avsg.2013.01.010"},{"key":"e_1_3_5_57_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1749-6632.2009.05032.x"},{"key":"e_1_3_5_58_2","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/21.8.1939"},{"key":"e_1_3_5_59_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI18420"},{"key":"e_1_3_5_60_2","doi-asserted-by":"publisher","DOI":"10.1096\/fj.06-6568fje"},{"key":"e_1_3_5_61_2","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvp152"},{"key":"e_1_3_5_62_2","doi-asserted-by":"publisher","DOI":"10.1159\/000189797"},{"key":"e_1_3_5_63_2","doi-asserted-by":"publisher","DOI":"10.2174\/138920107779941408"}],"container-title":["Journal of the American Heart Association"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.ahajournals.org\/doi\/full\/10.1161\/JAHA.115.001815","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,13]],"date-time":"2024-05-13T10:53:04Z","timestamp":1715597584000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ahajournals.org\/doi\/10.1161\/JAHA.115.001815"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,6,13]]},"references-count":62,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2016,6,13]]}},"alternative-id":["10.1161\/JAHA.115.001815"],"URL":"https:\/\/doi.org\/10.1161\/jaha.115.001815","relation":{},"ISSN":["2047-9980"],"issn-type":[{"value":"2047-9980","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,6,13]]},"assertion":[{"value":"2016-03-24","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2016-04-19","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2016-05-26","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e001815"}}