{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T06:58:32Z","timestamp":1792047512154,"version":"4.4.0"},"reference-count":46,"publisher":"American Society of Hematology","issue":"17","content-domain":{"domain":["ashpublications.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2021,10,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Thalassemia or sickle cell patients with hereditary persistence of fetal hemoglobin (HbF) have an ameliorated clinical phenotype and, in some cases, can achieve transfusion independence. Inactivation via genome editing of \u03b3-globin developmental suppressors, such as BCL11A or LRF\/ZBTB7A, or of their binding sites, have been shown to significantly increase expression of endogenous HbF. To broaden the therapeutic window beyond a single-editing approach, we have explored combinations of cis- and trans-editing targets to enhance HbF reactivation. Multiplex mutagenesis in adult CD34+ cells was well tolerated and did not lead to any detectable defect in the cells\u2019 proliferation and differentiation, either in vitro or in vivo. The combination of 1 trans and 1 cis mutation resulted in high editing retention in vivo, coupled with almost pancellular HbF expression in NBSGW mice. The greater in vivo performance of this combination was also recapitulated using a novel helper-dependent adenoviral-CRISPR vector (HD-Ad-dualCRISPR) in CD34+ cells from \u03b2-thalassemia patients transplanted to NBSGW mice. A pronounced increase in HbF expression was observed in human red blood cells in mice with established predominant \u03b20\/\u03b20-thalassemic hemopoiesis after in vivo injection of the HD-Ad-dualCRISPR vector. Collectively, our data suggest that the combination of cis and trans fetal globin reactivation mutations has the potential to significantly increase HbF both totally and on a per cell basis over single editing and could thus provide significant clinical benefit to patients with severe \u03b2-globin phenotype.<\/jats:p>","DOI":"10.1182\/blood.2020010020","type":"journal-article","created":{"date-parts":[[2021,6,4]],"date-time":"2021-06-04T20:12:38Z","timestamp":1622837558000},"page":"1540-1553","update-policy":"https:\/\/doi.org\/10.1182\/blood.2019cm0000","source":"Crossref","is-referenced-by-count":48,"title":["Enhanced HbF reactivation by multiplex mutagenesis of thalassemic CD34+ cells in vitro and in vivo"],"prefix":"10.1182","volume":"138","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3139-057X","authenticated-orcid":false,"given":"Nikoletta","family":"Psatha","sequence":"first","affiliation":[{"name":"Altius Institute for Biomedical Sciences, Seattle, WA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9906-4797","authenticated-orcid":false,"given":"Aphrodite","family":"Georgakopoulou","sequence":"additional","affiliation":[{"name":"School of Biology, Aristotle University of Thessaloniki, Thessaloniki, Greece;"},{"name":"Gene and Cell Therapy Center, Hematology Department-Hematopoietic Cell Transplantation Unit, George Papanikolaou Hospital, Thessaloniki, Greece; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chang","family":"Li","sequence":"additional","affiliation":[{"name":"Division of Medical Genetics and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vivek","family":"Nandakumar","sequence":"additional","affiliation":[{"name":"Altius Institute for Biomedical Sciences, Seattle, WA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grigorios","family":"Georgolopoulos","sequence":"additional","affiliation":[{"name":"Altius Institute for Biomedical Sciences, Seattle, WA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reyes","family":"Acosta","sequence":"additional","affiliation":[{"name":"Altius Institute for Biomedical Sciences, Seattle, WA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kiriaki","family":"Paschoudi","sequence":"additional","affiliation":[{"name":"School of Biology, Aristotle University of Thessaloniki, Thessaloniki, Greece;"},{"name":"Gene and Cell Therapy Center, Hematology Department-Hematopoietic Cell Transplantation Unit, George Papanikolaou Hospital, Thessaloniki, Greece; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2846-2007","authenticated-orcid":false,"given":"Jemma","family":"Nelson","sequence":"additional","affiliation":[{"name":"Altius Institute for Biomedical Sciences, Seattle, WA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Chee","sequence":"additional","affiliation":[{"name":"Altius Institute for Biomedical Sciences, Seattle, WA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anastasia","family":"Athanasiadou","sequence":"additional","affiliation":[{"name":"Gene and Cell Therapy Center, Hematology Department-Hematopoietic Cell Transplantation Unit, George Papanikolaou Hospital, Thessaloniki, Greece; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anastasia","family":"Kouvatsi","sequence":"additional","affiliation":[{"name":"School of Biology, Aristotle University of Thessaloniki, Thessaloniki, Greece;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1062-5296","authenticated-orcid":false,"given":"Alister Peter William","family":"Funnell","sequence":"additional","affiliation":[{"name":"Altius Institute for Biomedical Sciences, Seattle, WA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6742-6266","authenticated-orcid":false,"given":"Andr\u00e9","family":"Lieber","sequence":"additional","affiliation":[{"name":"Division of Medical Genetics and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evangelia","family":"Yannaki","sequence":"additional","affiliation":[{"name":"Gene and Cell Therapy Center, Hematology Department-Hematopoietic Cell Transplantation Unit, George Papanikolaou Hospital, Thessaloniki, Greece; and"},{"name":"Division of Hematology, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thalia","family":"Papayannopoulou","sequence":"additional","affiliation":[{"name":"Division of Hematology, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"234","reference":[{"issue":"3","key":"2022030115110070500_B1","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1182\/blood.V24.3.223.223","article-title":"Hereditary persistence of fetal hemoglobin in Greece. A study and a comparison","volume":"24","author":"Fessas","year":"1964","journal-title":"Blood."},{"issue":"10","key":"2022030115110070500_B2","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1038\/nmeth.3554","article-title":"Functional footprinting of regulatory DNA","volume":"12","author":"Vierstra","year":"2015","journal-title":"Nat Methods."},{"issue":"7577","key":"2022030115110070500_B3","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1038\/nature15521","article-title":"BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis","volume":"527","author":"Canver","year":"2015","journal-title":"Nature."},{"key":"2022030115110070500_B4","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/j.omtm.2018.08.003","article-title":"Disruption of the BCL11A erythroid enhancer reactivates fetal hemoglobin in erythroid cells of patients with \u03b2-thalassemia major","volume":"10","author":"Psatha","year":"2018","journal-title":"Mol Ther Methods Clin Dev."},{"issue":"5","key":"2022030115110070500_B5","doi-asserted-by":"crossref","first-page":"776","DOI":"10.1038\/s41591-019-0401-y","article-title":"Highly efficient therapeutic gene editing of human hematopoietic stem cells","volume":"25","author":"Wu","year":"2019","journal-title":"Nat Med."},{"issue":"6270","key":"2022030115110070500_B6","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1126\/science.aad3312","article-title":"Transcription factors LRF and BCL11A independently repress expression of fetal hemoglobin","volume":"351","author":"Masuda","year":"2016","journal-title":"Science."},{"issue":"4","key":"2022030115110070500_B7","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1038\/s41588-018-0085-0","article-title":"Natural regulatory mutations elevate the fetal globin gene via disruption of BCL11A or ZBTB7A binding","volume":"50","author":"Martyn","year":"2018","journal-title":"Nat Genet."},{"issue":"2","key":"2022030115110070500_B8","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1016\/j.cell.2018.03.016","article-title":"Direct promoter repression by BCL11A controls the fetal to adult hemoglobin switch","volume":"173","author":"Liu","year":"2018","journal-title":"Cell."},{"issue":"9","key":"2022030115110070500_B9","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1038\/nm.4170","article-title":"A genome-editing strategy to treat \u03b2-hemoglobinopathies that recapitulates a mutation associated with a benign genetic condition","volume":"22","author":"Traxler","year":"2016","journal-title":"Nat Med."},{"issue":"26","key":"2022030115110070500_B10","doi-asserted-by":"crossref","first-page":"2915","DOI":"10.1182\/blood-2018-03-838540","article-title":"Reactivation of \u03b3-globin in adult \u03b2-YAC mice after ex vivo and in vivo hematopoietic stem cell genome editing","volume":"131","author":"Li","year":"2018","journal-title":"Blood."},{"key":"2022030115110070500_B11","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/j.omtm.2018.12.008","article-title":"TALEN-mediated gene editing of HBG in human hematopoietic stem cells leads to therapeutic fetal hemoglobin induction","volume":"12","author":"Lux","year":"2018","journal-title":"Mol Ther Methods Clin Dev."},{"issue":"10","key":"2022030115110070500_B12","doi-asserted-by":"crossref","first-page":"1852","DOI":"10.1002\/sctm.17-0048","article-title":"Brief report: a differential transcriptomic profile of ex vivo expanded adult human hematopoietic stem cells empowers them for engraftment better than their surface phenotype","volume":"6","author":"Psatha","year":"2017","journal-title":"Stem Cells Transl Med."},{"key":"2022030115110070500_B13","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1007\/978-1-59745-060-7_8","article-title":"Ex vivo generation of human red blood cells: a new advance in stem cell engineering","volume":"482","author":"Douay","year":"2009","journal-title":"Methods Mol Biol."},{"issue":"2","key":"2022030115110070500_B14","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.stemcr.2014.12.005","article-title":"Nonirradiated NOD,B6.SCID Il2r\u03b3-\/- Kit(W41\/W41) (NBSGW) mice support multilineage engraftment of human hematopoietic cells","volume":"4","author":"McIntosh","year":"2015","journal-title":"Stem Cell Reports."},{"issue":"1","key":"2022030115110070500_B15","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1038\/mt.2011.195","article-title":"Hematopoietic stem cell mobilization for gene therapy of adult patients with severe \u03b2-thalassemia: results of clinical trials using G-CSF or plerixafor in splenectomized and nonsplenectomized subjects","volume":"20","author":"Yannaki","year":"2012","journal-title":"Mol Ther."},{"issue":"10","key":"2022030115110070500_B16","doi-asserted-by":"crossref","first-page":"852","DOI":"10.1089\/hum.2013.163","article-title":"Hematopoietic stem cell mobilization for gene therapy: superior mobilization by the combination of granulocyte-colony stimulating factor plus plerixafor in patients with \u03b2-thalassemia major","volume":"24","author":"Yannaki","year":"2013","journal-title":"Hum Gene Ther."},{"issue":"6","key":"2022030115110070500_B17","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1089\/hgtb.2014.005","article-title":"Superior long-term repopulating capacity of G-CSF+plerixafor-mobilized blood: implications for stem cell gene therapy by studies in the Hbb(th-3) mouse model","volume":"25","author":"Psatha","year":"2014","journal-title":"Hum Gene Ther Methods."},{"issue":"22","key":"2022030115110070500_B18","doi-asserted-by":"crossref","first-page":"10635","DOI":"10.1093\/nar\/16.22.10635","article-title":"Distal CCAAT box deletion in the A \u03b3 globin gene of two black adolescents with elevated fetal A \u03b3 globin","volume":"16","author":"Gilman","year":"1988","journal-title":"Nucleic Acids Res."},{"issue":"21","key":"2022030115110070500_B19","doi-asserted-by":"crossref","first-page":"3379","DOI":"10.1182\/bloodadvances.2019000820","article-title":"Genome editing of HBG1 and HBG2 to induce fetal hemoglobin","volume":"3","author":"M\u00e9tais","year":"2019","journal-title":"Blood Adv."},{"issue":"2","key":"2022030115110070500_B20","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1111\/j.1365-2141.2011.08710.x","article-title":"Kr\u00fcppel-like factor 4 activates HBG gene expression in primary erythroid cells","volume":"154","author":"Kalra","year":"2011","journal-title":"Br J Haematol."},{"issue":"4","key":"2022030115110070500_B21","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1016\/j.stem.2019.02.019","article-title":"Precise gene editing preserves hematopoietic stem cell function following transient p53-mediated DNA damage response","volume":"24","author":"Schiroli","year":"2019","journal-title":"Cell Stem Cell."},{"issue":"18","key":"2022030115110070500_B22","doi-asserted-by":"crossref","first-page":"2206","DOI":"10.1182\/blood-2016-04-711580","article-title":"In vivo transduction of primitive mobilized hematopoietic stem cells after intravenous injection of integrating adenovirus vectors","volume":"128","author":"Richter","year":"2016","journal-title":"Blood."},{"key":"2022030115110070500_B23","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1016\/j.omtm.2018.02.004","article-title":"Integrating HDAd5\/35++ vectors as a new platform for HSC gene therapy of hemoglobinopathies","volume":"9","author":"Li","year":"2018","journal-title":"Mol Ther Methods Clin Dev."},{"issue":"2","key":"2022030115110070500_B24","doi-asserted-by":"crossref","first-page":"598","DOI":"10.1172\/JCI122836","article-title":"In vivo hematopoietic stem cell gene therapy ameliorates murine thalassemia intermedia","volume":"129","author":"Wang","year":"2019","journal-title":"J Clin Invest."},{"key":"2022030115110070500_B25","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1016\/j.omtm.2018.04.008","article-title":"HDAd5\/35++ adenovirus vector expressing anti-CRISPR peptides decreases CRISPR\/Cas9 toxicity in human hematopoietic stem cells","volume":"9","author":"Li","year":"2018","journal-title":"Mol Ther Methods Clin Dev."},{"issue":"3","key":"2022030115110070500_B26","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1089\/hum.2009.077","article-title":"Mobilization of hematopoietic stem cells in a thalassemic mouse model: implications for human gene therapy of thalassemia","volume":"21","author":"Yannaki","year":"2010","journal-title":"Hum Gene Ther."},{"issue":"5826","key":"2022030115110070500_B27","doi-asserted-by":"crossref","first-page":"860","DOI":"10.1126\/science.1140881","article-title":"Immunology: Regulation of B versus T lymphoid lineage fate decision by the proto-oncogene LRF","volume":"316","author":"Maeda","year":"2007","journal-title":"Science."},{"issue":"15","key":"2022030115110070500_B28","doi-asserted-by":"crossref","first-page":"2845","DOI":"10.1182\/blood-2012-11-292037","article-title":"Role of LRF\/Pokemon in lineage fate decisions","volume":"121","author":"Lunardi","year":"2013","journal-title":"Blood."},{"issue":"13","key":"2022030115110070500_B29","doi-asserted-by":"crossref","first-page":"2467","DOI":"10.1084\/jem.20121846","article-title":"Bcl11a is essential for lymphoid development and negatively regulates p53","volume":"209","author":"Yu","year":"2012","journal-title":"J Exp Med."},{"issue":"6","key":"2022030115110070500_B30","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1038\/ni925","article-title":"Bcl11a is essential for normal lymphoid development","volume":"4","author":"Liu","year":"2003","journal-title":"Nat Immunol."},{"issue":"6","key":"2022030115110070500_B31","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1182\/blood-2012-03-418103","article-title":"LRF-mediated Dll4 repression in erythroblasts is necessary for hematopoietic stem cell maintenance","volume":"121","author":"Lee","year":"2013","journal-title":"Blood."},{"issue":"7","key":"2022030115110070500_B32","doi-asserted-by":"crossref","first-page":"eaay9392","DOI":"10.1126\/sciadv.aay9392","article-title":"Editing a \u03b3-globin repressor binding site restores fetal hemoglobin synthesis and corrects the sickle cell disease phenotype","volume":"6","author":"Weber","year":"2020","journal-title":"Sci Adv."},{"issue":"4","key":"2022030115110070500_B33","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1182\/blood-2013-09-528067","article-title":"Fetal hemoglobin in sickle cell anemia: a glass half full?","volume":"123","author":"Steinberg","year":"2014","journal-title":"Blood."},{"issue":"18","key":"2022030115110070500_B34","doi-asserted-by":"crossref","first-page":"4560","DOI":"10.1182\/bloodadvances.2020002475","article-title":"HRI depletion cooperates with pharmacologic inducers to elevate fetal hemoglobin and reduce sickle cell formation","volume":"4","author":"Peslak","year":"2020","journal-title":"Blood Adv."},{"issue":"10","key":"2022030115110070500_B35","doi-asserted-by":"crossref","first-page":"2817","DOI":"10.1182\/blood-2010-08-300723","article-title":"Therapeutic levels of fetal hemoglobin in erythroid progeny of \u03b2-thalassemic CD34+ cells after lentiviral vector-mediated gene transfer","volume":"117","author":"Wilber","year":"2011","journal-title":"Blood."},{"issue":"1","key":"2022030115110070500_B36","doi-asserted-by":"crossref","first-page":"6732","DOI":"10.1038\/s41598-017-06757-0","article-title":"Targeted DNA methylation in human cells using engineered dCas9-methyltransferases","volume":"7","author":"Xiong","year":"2017","journal-title":"Sci Rep."},{"issue":"9","key":"2022030115110070500_B37","doi-asserted-by":"crossref","first-page":"2255","DOI":"10.1158\/1078-0432.CCR-16-1300","article-title":"Multiplex genome editing to generate universal CAR T cells resistant to PD1 inhibition","volume":"23","author":"Ren","year":"2017","journal-title":"Clin Cancer Res."},{"issue":"1","key":"2022030115110070500_B38","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1038\/cr.2016.142","article-title":"CRISPR-Cas9-mediated multiplex gene editing in CAR-T cells","volume":"27","author":"Liu","year":"2017","journal-title":"Cell Res."},{"issue":"10","key":"2022030115110070500_B39","doi-asserted-by":"crossref","first-page":"17002","DOI":"10.18632\/oncotarget.15218","article-title":"A versatile system for rapid multiplex genome-edited CAR T cell generation","volume":"8","author":"Ren","year":"2017","journal-title":"Oncotarget."},{"issue":"12","key":"2022030115110070500_B40","first-page":"7636","article-title":"Developmental regulation of human gamma-globin genes in transgenic mice","volume":"13","author":"Stamatoyannopoulos","year":"1993","journal-title":"Mol Cell Biol."},{"issue":"12","key":"2022030115110070500_B41","doi-asserted-by":"crossref","first-page":"6677","DOI":"10.1172\/JCI140189","article-title":"BCL11A enhancer-edited hematopoietic stem cells persist in rhesus monkeys without toxicity","volume":"130","author":"Demirci","year":"2020","journal-title":"J\u00a0Clin Invest."},{"key":"2022030115110070500_B42","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.omtm.2017.11.004","article-title":"A combined in vivo HSC transduction\/selection approach results in efficient and stable gene\u00a0expression in peripheral blood cells in mice","volume":"8","author":"Wang","year":"2017","journal-title":"Mol Ther Methods Clin Dev."},{"issue":"2","key":"2022030115110070500_B43","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1016\/j.ymthe.2020.09.001","article-title":"In vivo HSC gene therapy using a bi-modular HDAd5\/35++ vector cures sickle cell disease in a mouse model","volume":"29","author":"Li","year":"2021","journal-title":"Mol Ther."},{"issue":"12","key":"2022030115110070500_B44","doi-asserted-by":"crossref","first-page":"2195","DOI":"10.1016\/j.ymthe.2019.08.006","article-title":"Targeted integration and high-level transgene expression in AAVS1 transgenic mice after in vivo HSC transduction with HDAd5\/35++ vectors","volume":"27","author":"Li","year":"2019","journal-title":"Mol Ther."},{"issue":"15","key":"2022030115110070500_B45","doi-asserted-by":"crossref","first-page":"8263","DOI":"10.1128\/JVI.77.15.8263-8271.2003","article-title":"Replication-deficient human adenovirus type 35 vectors for gene transfer and vaccination: efficient human cell infection and bypass of preexisting adenovirus immunity","volume":"77","author":"Vogels","year":"2003","journal-title":"J Virol."},{"issue":"4","key":"2022030115110070500_B46","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1038\/mt.2008.19","article-title":"Transduction properties of adenovirus serotype 35 vectors after intravenous administration into nonhuman primates","volume":"16","author":"Sakurai","year":"2008","journal-title":"Mol Ther."}],"updated-by":[{"DOI":"10.1182\/blood.2022015968","type":"erratum","label":"Erratum","source":"publisher","updated":{"date-parts":[[2022,3,31]],"date-time":"2022-03-31T00:00:00Z","timestamp":1648684800000}}],"container-title":["Blood"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/ashpublications.org\/blood\/article-pdf\/138\/17\/1540\/1876977\/bloodbld2020010020.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/ashpublications.org\/blood\/article-pdf\/138\/17\/1540\/1876977\/bloodbld2020010020.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,3,1]],"date-time":"2022-03-01T15:11:45Z","timestamp":1646147505000},"score":1,"resource":{"primary":{"URL":"https:\/\/ashpublications.org\/blood\/article\/138\/17\/1540\/476068\/Enhanced-HbF-reactivation-by-multiplex-mutagenesis"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,28]]},"references-count":46,"journal-issue":{"issue":"17","published-print":{"date-parts":[[2021,10,28]]}},"URL":"https:\/\/doi.org\/10.1182\/blood.2020010020","relation":{"erratum":[{"id-type":"doi","id":"10.1182\/blood.2022015968","asserted-by":"object"}]},"ISSN":["0006-4971","1528-0020"],"issn-type":[{"value":"0006-4971","type":"print"},{"value":"1528-0020","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,28]]}}}