Yanan Zhang, Dingyue Tang, Li Wang, Jing Yang, Xia Wu, Xiao Xiao, Jian-She Wang
{"title":"用腺相关病毒载体递送Zfyve19预防Zfyve19-/-小鼠模型中门脉纤维化。","authors":"Yanan Zhang, Dingyue Tang, Li Wang, Jing Yang, Xia Wu, Xiao Xiao, Jian-She Wang","doi":"10.1089/hum.2023.041","DOIUrl":null,"url":null,"abstract":"<p><p>Zinc finger FYVE-type containing 19 (ZFYVE19) deficiency, caused by biallelic <i>ZFYVE19</i> complete loss-of-function variants, is a recently identified chronic hepatobiliary disorder characterized by obvious portal-tract fibrosis, increased numbers of bile ducts with malformations, and abnormal levels of serum markers of hepatobiliary injury. As liver-targeted adeno-associated virus (AAV) gene therapy has been used successfully in hepatobiliary diseases, liver-targeted gene therapy has been explored in a mouse model of this disorder. Three <i>ZFYVE19</i> AAV vectors (AAV-<i>hZFYVE19</i>, AAV-<i>hZFYVE19-m</i>, and AAV-<i>hZFYVE19-co</i>) were constructed and injected into <i>Zfyve19<sup>-/-</sup></i> mice, which were treated with alpha-naphthyl isothiocyanate, a hepatobiliary toxin. Hematoxylin/eosin, immunohistochemical staining, immunofluorescence staining, Sirius Red staining, real-time quantitative PCR, and Western blotting of liver tissue, along with serum hepatobiliary injury marker analyses, were performed to evaluate the effects of gene therapy. AAV-<i>hZFYVE19</i> decreased serum hepatobiliary injury markers, portal-tract inflammation, ductal hyperplasia, and portal-tract fibrosis in the <i>Zfyve19<sup>-/-</sup></i> model mice most substantially at a relatively low dose (1 × 10<sup>11</sup> vg/kg), whereas AAV-<i>hZFYVE19</i> at a higher dose gradually lost the abovementioned benefits and even caused deterioration at the highest dose of 5 × 10<sup>12</sup> vg/kg. These observations verified the pathogenicity of ZFYVE19 deficiency and suggested that the <i>ZFYVE19</i> gene needs to function well at an optimal level of expression; both too low and too high a <i>ZFYVE19</i> expression may be harmful.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":null,"pages":null},"PeriodicalIF":3.9000,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Prevention of Portal-Tract Fibrosis in <i>Zfyve19<sup>-/-</sup></i> Mouse Model with Adeno-Associated Virus Vector Delivering <i>ZFYVE19</i>.\",\"authors\":\"Yanan Zhang, Dingyue Tang, Li Wang, Jing Yang, Xia Wu, Xiao Xiao, Jian-She Wang\",\"doi\":\"10.1089/hum.2023.041\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Zinc finger FYVE-type containing 19 (ZFYVE19) deficiency, caused by biallelic <i>ZFYVE19</i> complete loss-of-function variants, is a recently identified chronic hepatobiliary disorder characterized by obvious portal-tract fibrosis, increased numbers of bile ducts with malformations, and abnormal levels of serum markers of hepatobiliary injury. As liver-targeted adeno-associated virus (AAV) gene therapy has been used successfully in hepatobiliary diseases, liver-targeted gene therapy has been explored in a mouse model of this disorder. Three <i>ZFYVE19</i> AAV vectors (AAV-<i>hZFYVE19</i>, AAV-<i>hZFYVE19-m</i>, and AAV-<i>hZFYVE19-co</i>) were constructed and injected into <i>Zfyve19<sup>-/-</sup></i> mice, which were treated with alpha-naphthyl isothiocyanate, a hepatobiliary toxin. Hematoxylin/eosin, immunohistochemical staining, immunofluorescence staining, Sirius Red staining, real-time quantitative PCR, and Western blotting of liver tissue, along with serum hepatobiliary injury marker analyses, were performed to evaluate the effects of gene therapy. AAV-<i>hZFYVE19</i> decreased serum hepatobiliary injury markers, portal-tract inflammation, ductal hyperplasia, and portal-tract fibrosis in the <i>Zfyve19<sup>-/-</sup></i> model mice most substantially at a relatively low dose (1 × 10<sup>11</sup> vg/kg), whereas AAV-<i>hZFYVE19</i> at a higher dose gradually lost the abovementioned benefits and even caused deterioration at the highest dose of 5 × 10<sup>12</sup> vg/kg. These observations verified the pathogenicity of ZFYVE19 deficiency and suggested that the <i>ZFYVE19</i> gene needs to function well at an optimal level of expression; both too low and too high a <i>ZFYVE19</i> expression may be harmful.</p>\",\"PeriodicalId\":13007,\"journal\":{\"name\":\"Human gene therapy\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.9000,\"publicationDate\":\"2023-12-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Human gene therapy\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1089/hum.2023.041\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2023/10/18 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Human gene therapy","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1089/hum.2023.041","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/10/18 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
Prevention of Portal-Tract Fibrosis in Zfyve19-/- Mouse Model with Adeno-Associated Virus Vector Delivering ZFYVE19.
Zinc finger FYVE-type containing 19 (ZFYVE19) deficiency, caused by biallelic ZFYVE19 complete loss-of-function variants, is a recently identified chronic hepatobiliary disorder characterized by obvious portal-tract fibrosis, increased numbers of bile ducts with malformations, and abnormal levels of serum markers of hepatobiliary injury. As liver-targeted adeno-associated virus (AAV) gene therapy has been used successfully in hepatobiliary diseases, liver-targeted gene therapy has been explored in a mouse model of this disorder. Three ZFYVE19 AAV vectors (AAV-hZFYVE19, AAV-hZFYVE19-m, and AAV-hZFYVE19-co) were constructed and injected into Zfyve19-/- mice, which were treated with alpha-naphthyl isothiocyanate, a hepatobiliary toxin. Hematoxylin/eosin, immunohistochemical staining, immunofluorescence staining, Sirius Red staining, real-time quantitative PCR, and Western blotting of liver tissue, along with serum hepatobiliary injury marker analyses, were performed to evaluate the effects of gene therapy. AAV-hZFYVE19 decreased serum hepatobiliary injury markers, portal-tract inflammation, ductal hyperplasia, and portal-tract fibrosis in the Zfyve19-/- model mice most substantially at a relatively low dose (1 × 1011 vg/kg), whereas AAV-hZFYVE19 at a higher dose gradually lost the abovementioned benefits and even caused deterioration at the highest dose of 5 × 1012 vg/kg. These observations verified the pathogenicity of ZFYVE19 deficiency and suggested that the ZFYVE19 gene needs to function well at an optimal level of expression; both too low and too high a ZFYVE19 expression may be harmful.
期刊介绍:
Human Gene Therapy is the premier, multidisciplinary journal covering all aspects of gene therapy. The Journal publishes in-depth coverage of DNA, RNA, and cell therapies by delivering the latest breakthroughs in research and technologies. Human Gene Therapy provides a central forum for scientific and clinical information, including ethical, legal, regulatory, social, and commercial issues, which enables the advancement and progress of therapeutic procedures leading to improved patient outcomes, and ultimately, to curing diseases.