用腺相关病毒载体递送Zfyve19预防Zfyve19-/-小鼠模型中门脉纤维化。

IF 3.9 3区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Human gene therapy Pub Date : 2023-12-01 Epub Date: 2023-10-18 DOI:10.1089/hum.2023.041
Yanan Zhang, Dingyue Tang, Li Wang, Jing Yang, Xia Wu, Xiao Xiao, Jian-She Wang
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引用次数: 0

摘要

含锌指FYVE 19型(ZFYVE19)缺乏症是由双等位基因ZFYVE-19功能完全丧失变体引起的,是一种最近发现的慢性肝胆疾病,其特征是明显的门脉纤维化、伴有畸形的胆管数量增加以及肝胆损伤的血清标志物水平异常。由于肝靶向腺相关病毒(AAV)基因治疗已成功用于肝胆疾病,因此已在该疾病的小鼠模型中探索肝靶向基因治疗。构建了三种ZFYVE19-AAV载体(AAV-hZFYVE19、AAV-hZFYVE19-m和AAV-hz FYVE19-co),并将其注射到用肝胆毒素α-萘异硫氰酸酯处理的ZFYVE19-/-小鼠中。对肝组织进行苏木精/伊红、免疫组织化学染色、免疫荧光染色、天狼星红染色、实时定量PCR和Western印迹,以及血清肝胆损伤标志物分析,以评估基因治疗的效果。AAV-hZFYVE19在相对较低的剂量下显著降低了Zfyve19-/-模型小鼠的血清肝胆损伤标志物、门脉炎症、导管增生和门脉纤维化(1 × 1011vg/kg),而较高剂量的AAV-hZFYVE19逐渐失去上述益处,甚至在最高剂量5 × 1012伏/公斤。这些观察结果证实了ZFYVE19缺乏症的致病性,并表明ZFYVE-19基因需要在最佳表达水平下发挥良好的功能;ZFYVE19的表达过低和过高都可能是有害的。
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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.

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来源期刊
Human gene therapy
Human gene therapy 医学-生物工程与应用微生物
CiteScore
6.50
自引率
4.80%
发文量
131
审稿时长
4-8 weeks
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.
期刊最新文献
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