Endoscopic-mediated, biliary hydrodynamic injection mediating clinically relevant levels of gene delivery in pig liver.
Endoscopic-mediated, biliary hydrodynamic injection mediating clinically relevant levels of gene delivery in pig liver.
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DOI：
10.1016/j.gie.2021.06.016
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发表时间：
2021-12
影响因子：
7.7
通讯作者：
Kumbhari V
中科院分区：
文献类型：
--
作者：
Kruse RL;Huang Y;Shum T;Bai L;Ding H;Wang ZZ;Selaru FM;Kumbhari V
基因治疗可以为许多遗传性单基因肝病提供治疗方法。临床试验主要集中在用于肝脏基因传递的腺相关病毒(AAV)。然而，这些载体受到包装尺寸小、衣壳免疫反应和不能重新给药的限制。作为一种替代方案，经血管途径的非病毒流体动力学注射可以成功地将质粒DNA(PDNA)输送到小鼠肝脏，但在大型动物模型中取得的成功有限。我们使用ERCP和动力注射器来研究通过胆道系统将PDNA流体动力输送到猪的肝脏中。血友病B缺乏的人凝血因子IX(HFIX)被用作模型基因治疗。胆道流体动力注射耐受性良好，生命体征、肝酶、血液学或组织学均无明显改变。聚合酶链式反应未检测到非靶向其他器官的PDNA传递。免疫组织化学结果显示，注射5.5mgPDNA后，50.19%的大鼠肝脏hFIX染色阳性，所有肝小叶均有hFIX阳性表达。HFIX阳性肝细胞集中在中央静脉周围，向外辐射至所有3个代谢区。猪胆管注射的转染率显著高于小鼠血管注射(32.7-51.9%vs18.9%，p<；0.0001)。在临床相关的人类大小的大型动物中，通过ERCP进行胆道流体动力注射可以获得比AAV更高的肝细胞转染率，而且成本更低。这项技术可能为人类肝病的基因治疗提供一个平台。
Gene therapy could provide curative therapies to many inherited monogenic liver diseases. Clinical trials have largely focused on adeno-associated viruses (AAV) for liver gene delivery. These vectors, however, are limited by small packaging size, capsid immune responses, and inability to re-dose. As an alternative, nonviral, hydrodynamic injection through vascular routes can successfully deliver plasmid DNA (pDNA) into mouse liver but has achieved limited success in large animal models. We explored hydrodynamic delivery of pDNA through the biliary system into the liver of pigs using ERCP, and a power injector to supply hydrodynamic force. Human Factor IX (hFIX), deficient in Hemophilia B, was used as a model gene therapy. Biliary hydrodynamic injection was well tolerated without significant changes in vital signs, liver enzymes, hematology, or histology. No off-target pDNA delivery to other organs was detected by PCR. Immunohistochemistry revealed that 50.19% of the liver stained positive for hFIX after hydrodynamic injection at 5.5 mg pDNA, with every hepatic lobule in all liver lobes demonstrating hFIX-expression. hFIX-positive hepatocytes were concentrated around the central vein, radiating outward across all 3 metabolic zones. Biliary hydrodynamic injection in pigs resulted in significantly higher transfection efficiency than mouse vascular hydrodynamic injection at matched pDNA per liver weight dose (32.7-51.9% vs 18.9%, p<0.0001). Biliary hydrodynamic injection via ERCP can achieve higher transfection efficiency into hepatocytes compared with AAV at magnitudes less cost in a clinically relevant human-sized large animal. This technology may serve as a platform for gene therapy of human liver diseases.
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DOI：
10.1038/mtna.2016.63
发表时间：
2016-08-30
期刊：
Molecular therapy. Nucleic acids
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.7
作者：
KAWABATA, K;TAKAKURA, Y;HASHIDA, M
通讯作者：
HASHIDA, M
影响因子：
12.4
作者：
Kamimura, Kenya;Suda, Takeshi;Liu, Dexi
通讯作者：
Liu, Dexi
影响因子：
--
作者：
Calcedo, Roberto;Morizono, Hiroki;Wilson, James M.
通讯作者：
Wilson, James M.
影响因子：
24.5
作者：
Hu, J;Zhang, X;Fabre, JW
通讯作者：
Fabre, JW