{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,22]],"date-time":"2026-01-22T23:43:33Z","timestamp":1769125413829,"version":"3.49.0"},"reference-count":54,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"8","content-domain":{"domain":["www.ahajournals.org"],"crossmark-restriction":true},"short-container-title":["ATVB"],"published-print":{"date-parts":[[2023,8]]},"abstract":"<jats:sec>\n                    <jats:title>BACKGROUND:<\/jats:title>\n                    <jats:p>Specialized valve endothelial cell (VEC) populations are localized oriented to blood flow in developing aortic and mitral valves, but their roles in valve development and disease are unknown. In the aortic valve (AoV), a population of VECs on the fibrosa side expresses the transcription factor Prox1 together with genes found in lymphatic ECs. In this study, we examine Prox1\u2019s role in regulating a lymphatic-like gene network and promoting VEC diversity required for the development of the stratified trilaminar extracellular matrix (ECM) of murine AoV leaflets.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>METHODS:<\/jats:title>\n                    <jats:p>\n                      To determine whether disruption of Prox1 localization affects heart valve development, we generated mice (\n                      <jats:italic toggle=\"yes\">\n                        NFATc1\n                        <jats:sup>enCre<\/jats:sup>\n                      <\/jats:italic>\n                      <jats:italic toggle=\"yes\">Prox1<\/jats:italic>\n                      gain-of-function) in which Prox1 is overexpressed on the ventricularis side of the AoV beginning in embryonic development. To identify potential targets of Prox1, we performed cleavage under targets and release using nuclease on wild-type and\n                      <jats:italic toggle=\"yes\">\n                        NFATc1\n                        <jats:sup>enCre<\/jats:sup>\n                      <\/jats:italic>\n                      <jats:italic toggle=\"yes\">Prox1<\/jats:italic>\n                      gain-of-function AoVs with validation by colocalization in vivo using RNA in situ hybridization in\n                      <jats:italic toggle=\"yes\">\n                        NFATc1\n                        <jats:sup>enCre<\/jats:sup>\n                      <\/jats:italic>\n                      <jats:italic toggle=\"yes\">Prox1<\/jats:italic>\n                      gain-of-function AoVs. Natural induction of Prox1 and target gene expression was evaluated in myxomatous AoVs in a mouse model of Marfan syndrome (\n                      <jats:italic toggle=\"yes\">\n                        Fbn1\n                        <jats:sup>C1039G\/+<\/jats:sup>\n                      <\/jats:italic>\n                      ).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>RESULTS:<\/jats:title>\n                    <jats:p>\n                      The overexpression of Prox1 is sufficient to cause enlargement of AoVs by postnatal day (P)0, as well as a decrease in ventricularis-specific gene expression and disorganized interstitial ECM layers at P7. We identified potential targets of Prox1 known to play roles in lymphatic ECs including\n                      <jats:italic toggle=\"yes\">Flt1<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">Efnb2<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">Egfl7<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">Cx37<\/jats:italic>\n                      . Ectopic Prox1 colocalized with induced\n                      <jats:italic toggle=\"yes\">Flt1<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">Efnb2<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">Cx37<\/jats:italic>\n                      expression in\n                      <jats:italic toggle=\"yes\">\n                        NFATc1\n                        <jats:sup>enCre<\/jats:sup>\n                      <\/jats:italic>\n                      <jats:italic toggle=\"yes\">Prox1<\/jats:italic>\n                      gain-of-function AoVs. Moreover, in Marfan syndrome myxomatous AoVs, endogenous Prox1, and its identified targets, were ectopically induced in ventricularis side VECs.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>CONCLUSIONS:<\/jats:title>\n                    <jats:p>Our results support a role for Prox1 in localized lymphatic-like gene expression on the fibrosa side of the AoV. Furthermore, localized VEC specialization is required for development of the stratified trilaminar ECM critical for AoV function and is dysregulated in congenitally malformed valves.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1161\/atvbaha.123.319424","type":"journal-article","created":{"date-parts":[[2023,6,29]],"date-time":"2023-06-29T05:00:24Z","timestamp":1688014824000},"page":"1478-1493","update-policy":"https:\/\/doi.org\/10.1161\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":11,"title":["Localized Prox1 Regulates Aortic Valve Endothelial Cell Diversity and Extracellular Matrix Stratification in Mice"],"prefix":"10.1161","volume":"43","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4880-8394","authenticated-orcid":true,"given":"Anna","family":"O\u2019Donnell","sequence":"first","affiliation":[{"name":"Division of Molecular Cardiovascular Biology (A.O., B.A.G., R.W., C.M.A., C.O.S., D.P.M., K.E.Y.), Cincinnati Children\u2019s Hospital Medical Center, OH."},{"name":"Molecular and Developmental Biology Graduate Program (A.O., S.M.), University of Cincinnati, OH."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brittany A.","family":"Gonzalez","sequence":"additional","affiliation":[{"name":"Division of Molecular Cardiovascular Biology (A.O., B.A.G., R.W., C.M.A., C.O.S., D.P.M., K.E.Y.), Cincinnati Children\u2019s Hospital Medical Center, OH."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0694-024X","authenticated-orcid":true,"given":"Shreyasi","family":"Mukherjee","sequence":"additional","affiliation":[{"name":"Division of Developmental Biology (S.M., A.M.Z.), Cincinnati Children\u2019s Hospital Medical Center, OH."},{"name":"Molecular and Developmental Biology Graduate Program (A.O., S.M.), University of Cincinnati, OH."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruby","family":"Wilson","sequence":"additional","affiliation":[{"name":"Division of Molecular Cardiovascular Biology (A.O., B.A.G., R.W., C.M.A., C.O.S., D.P.M., K.E.Y.), Cincinnati Children\u2019s Hospital Medical Center, OH."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christina M.","family":"Alfieri","sequence":"additional","affiliation":[{"name":"Division of Molecular Cardiovascular Biology (A.O., B.A.G., R.W., C.M.A., C.O.S., D.P.M., K.E.Y.), Cincinnati Children\u2019s Hospital Medical Center, OH."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Casey O.","family":"Swoboda","sequence":"additional","affiliation":[{"name":"Division of Molecular Cardiovascular Biology (A.O., B.A.G., R.W., C.M.A., C.O.S., D.P.M., K.E.Y.), Cincinnati Children\u2019s Hospital Medical Center, OH."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Douglas P.","family":"Millay","sequence":"additional","affiliation":[{"name":"Division of Molecular Cardiovascular Biology (A.O., B.A.G., R.W., C.M.A., C.O.S., D.P.M., K.E.Y.), Cincinnati Children\u2019s Hospital Medical Center, OH."},{"name":"Department of Pediatrics, College of Medicine (D.P.M., A.M.Z., K.E.Y.), University of Cincinnati, OH."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3217-3590","authenticated-orcid":true,"given":"Aaron M.","family":"Zorn","sequence":"additional","affiliation":[{"name":"Division of Developmental Biology (S.M., A.M.Z.), Cincinnati Children\u2019s Hospital Medical Center, OH."},{"name":"Department of Pediatrics, College of Medicine (D.P.M., A.M.Z., K.E.Y.), University of Cincinnati, OH."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1444-7124","authenticated-orcid":true,"given":"Katherine E.","family":"Yutzey","sequence":"additional","affiliation":[{"name":"Division of Molecular Cardiovascular Biology (A.O., B.A.G., R.W., C.M.A., C.O.S., D.P.M., K.E.Y.), Cincinnati Children\u2019s Hospital Medical Center, OH."},{"name":"Department of 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