{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T18:12:20Z","timestamp":1790791940465,"version":"4.1.0"},"reference-count":40,"publisher":"Wiley","license":[{"start":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T00:00:00Z","timestamp":1760054400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["FOR2060"],"award-info":[{"award-number":["FOR2060"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["NI 1301\/8\u20101"],"award-info":[{"award-number":["NI 1301\/8\u20101"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bpspubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["British J Pharmacology"],"abstract":"<jats:sec><jats:title>Background and Purpose<\/jats:title><jats:p>Cyclic guanosine monophosphate (cGMP) is a ubiquitous second messenger involved in human (patho\u2010)physiology. Phosphodiesterase 5 (PDE5) is a major cGMP hydrolyzing enzyme in many cell types including vascular smooth muscle cells (VSMCs). Several highly selective PDE5 inhibitors are in clinical use. However, there are currently no tissue\/cell\u2010specific PDE5 inhibitors with precise action lacking systemic side effects. We sought to develop a new non\u2010chemical PDE5 inhibitor which could be used in specific tissues.<\/jats:p><\/jats:sec><jats:sec><jats:title>Experimental Approach<\/jats:title><jats:p>The <jats:bold>s<\/jats:bold>ingle\u2010chain variable fragment (scFv) construct was generated by sequencing heavy and light chains of the inhibitory antibody against GAF\u2010A domain of PDE5. The scFv was purified from bacteria and tested by competitive binding assays, and its sequence was expressed in aortic VSMCs using adenoviral gene transfer. Functional effects on cGMP levels were tested using live cell imaging of cGMP, phosphorylation of cGMP\u2010dependent protein kinase (PKG) substrate and by measuring vasorelaxation in mouse aortic rings following svFv expression.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Results<\/jats:title><jats:p>scFv expressed as a 29\u2010kDa protein and could specifically bind to the GAF\u2010A domain of PDE5. Live cell imaging demonstrated that scFv increased NO\u2010donor stimulated intracellular cGMP, whereas subsequent effects of the PDE5 inhibitor sildenafil were diminished, as was the PKG\u2010dependent VASP phosphorylation in response to sildenafil. Lastly, adenoviral expression of scFv in mouse aortic rings potentiated NO\u2010donor\u2010induced vasorelaxation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion and Implication<\/jats:title><jats:p>Our data suggest that recombinant scFv could represent a new class of intracellular PDE5 inhibitors. Treatment of chronic conditions with scFv could provide tissue\u2010\/cell\u2010specific therapy, minimizing potential side effects.<\/jats:p><\/jats:sec>","DOI":"10.1111\/bph.70221","type":"journal-article","created":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T12:55:38Z","timestamp":1760100938000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["An intracellular recombinant single\u2010chain variable antibody fragment as a new class of phosphodiesterase type 5 inhibitors"],"prefix":"10.1111","author":[{"given":"K\u00fcrsat","family":"Kirkg\u00f6z","sequence":"first","affiliation":[{"name":"Institute of Experimental Cardiovascular Research University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"},{"name":"German Center of Cardiovascular Research (DZHK) Partner Site Hamburg\/Kiel\/L\u00fcbeck  Hamburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophie","family":"Sprenger","sequence":"additional","affiliation":[{"name":"Institute of Experimental Cardiovascular Research University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"},{"name":"German Center of Cardiovascular Research (DZHK) Partner Site Hamburg\/Kiel\/L\u00fcbeck  Hamburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olga","family":"Schweigert","sequence":"additional","affiliation":[{"name":"German Center of Cardiovascular Research (DZHK) Partner Site Hamburg\/Kiel\/L\u00fcbeck  Hamburg Germany"},{"name":"Institute of Cardiogenetics University Hospital Schlewig\u2010Holstein, University of L\u00fcbeck  L\u00fcbeck Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sanika","family":"Mohagaonkar","sequence":"additional","affiliation":[{"name":"Institute of Experimental Cardiovascular Research University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"},{"name":"German Center of Cardiovascular Research (DZHK) Partner Site Hamburg\/Kiel\/L\u00fcbeck  Hamburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matti","family":"Walsdorff","sequence":"additional","affiliation":[{"name":"German Center of Cardiovascular Research (DZHK) Partner Site Hamburg\/Kiel\/L\u00fcbeck  Hamburg Germany"},{"name":"Department of Cardiology University Heart and Vascular Center Hamburg, University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanja","family":"Zeller","sequence":"additional","affiliation":[{"name":"German Center of Cardiovascular Research (DZHK) Partner Site Hamburg\/Kiel\/L\u00fcbeck  Hamburg Germany"},{"name":"Institute of Cardiogenetics University Hospital Schlewig\u2010Holstein, University of L\u00fcbeck  L\u00fcbeck Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Filip","family":"Berisha","sequence":"additional","affiliation":[{"name":"German Center of Cardiovascular Research (DZHK) Partner Site Hamburg\/Kiel\/L\u00fcbeck  Hamburg Germany"},{"name":"Department of Cardiology University Heart and Vascular Center Hamburg, University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Viacheslav O.","family":"Nikolaev","sequence":"additional","affiliation":[{"name":"Institute of Experimental Cardiovascular Research University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"},{"name":"German Center of Cardiovascular Research (DZHK) Partner Site Hamburg\/Kiel\/L\u00fcbeck  Hamburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sergei D.","family":"Rybalkin","sequence":"additional","affiliation":[{"name":"Institute of Experimental Cardiovascular Research University Medical Center Hamburg\u2010Eppendorf  Hamburg Germany"},{"name":"German Center of Cardiovascular Research (DZHK) Partner Site Hamburg\/Kiel\/L\u00fcbeck  Hamburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,10,10]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF03401832"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/bph.16181"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/bph.14205"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/14651858.CD012621.pub2"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvy221"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrm911"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.044537"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.3140379"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms19030801"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tips.2021.11.002"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/bph.17441"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2023.112433"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-050410-100030"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00210-019-01779-z"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvq410"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2025.1545044"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.109.890962"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2011.08.036"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.1515\/hsz-2019-0386"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.1126\/stke.19pe12"},{"key":"e_1_2_12_22_1","doi-asserted-by":"publisher","DOI":"10.1161\/hc4401.098432"},{"key":"e_1_2_12_23_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.2009.172189"},{"key":"e_1_2_12_24_1","doi-asserted-by":"publisher","DOI":"10.1161\/circresaha.117.303776"},{"key":"e_1_2_12_25_1","doi-asserted-by":"publisher","DOI":"10.1002\/1526-968x(200009)28:1<>3.0.co;2-c"},{"key":"e_1_2_12_26_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0303758"},{"key":"e_1_2_12_27_1","doi-asserted-by":"publisher","DOI":"10.3390\/jcdd5020020"},{"key":"e_1_2_12_28_1","doi-asserted-by":"publisher","DOI":"10.1111\/bph.15178"},{"key":"e_1_2_12_29_1","doi-asserted-by":"publisher","DOI":"10.1152\/jappl.2001.91.3.1421"},{"key":"e_1_2_12_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)52413-3"},{"key":"e_1_2_12_31_1","doi-asserted-by":"publisher","DOI":"10.1161\/01.CIR.0000094403.78467.C3"},{"key":"e_1_2_12_32_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.C400505200"},{"key":"e_1_2_12_33_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3000410"},{"key":"e_1_2_12_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jconrel.2017.02.003"},{"key":"e_1_2_12_35_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M106562200"},{"key":"e_1_2_12_36_1","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/cdg051"},{"key":"e_1_2_12_37_1","doi-asserted-by":"publisher","DOI":"10.1515\/hsz-2018-0155"},{"key":"e_1_2_12_38_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1813446"},{"key":"e_1_2_12_39_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.bioconjchem.9b00025"},{"key":"e_1_2_12_40_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-62703-459-3_6"},{"key":"e_1_2_12_41_1","doi-asserted-by":"publisher","DOI":"10.1002\/biot.201900430"}],"container-title":["British Journal of Pharmacology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/bpspubs.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/bph.70221","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T00:09:23Z","timestamp":1760486963000},"score":1,"resource":{"primary":{"URL":"https:\/\/bpspubs.onlinelibrary.wiley.com\/doi\/10.1111\/bph.70221"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,10]]},"references-count":40,"alternative-id":["10.1111\/bph.70221"],"URL":"https:\/\/doi.org\/10.1111\/bph.70221","archive":["Portico"],"relation":{},"ISSN":["0007-1188","1476-5381"],"issn-type":[{"value":"0007-1188","type":"print"},{"value":"1476-5381","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,10]]},"assertion":[{"value":"2025-02-27","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-09-10","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-10","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"bph.70221"}}