Delivery of non-viral naked DNA vectors to liver in small weaned pigs by hydrodynamic retrograde intrabiliary injection.
Delivery of non-viral naked DNA vectors to liver in small weaned pigs by hydrodynamic retrograde intrabiliary injection.
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DOI：
10.1016/j.omtm.2022.01.006
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发表时间：
2022-03-10
期刊：
影响因子：
--
通讯作者：
Thöny B
中科院分区：
文献类型：
--
作者：
Chan T;Grisch-Chan HM;Schmierer P;Subotic U;Rimann N;Scherer T;Hetzel U;Bozza M;Harbottle R;Williams JA;Steblaj B;Ringer SK;Häberle J;Sidler X;Thöny B
通过在新生儿中递送非整合治疗载体的肝脏基因治疗由于在生长的肝脏中稀释和丧失功效的风险而仍然具有挑战性。以前，我们报道了肝细胞转染仔猪门静脉注射裸DNA载体。在这里，我们建立了交付裸DNA载体，以靶向门静脉周围肝细胞在断奶仔猪的流体动力学逆行胆管内注射（HRII）。外科手术包括剖腹手术和短暂的肝脏隔离。对于载体递送，通过肠切开术将导管放置在胆总管内。在最佳条件下，加压注射后在肝组织中未观察到组织学异常。用从肝脏特异性启动子表达荧光素酶的载体观察到所有测试的肝脏样品中肝细胞的转染。然而，载体拷贝数和荧光素酶表达与流体动力学门静脉内注射相比较低。使用具有复制潜力的非整合裸DNA载体在猪中观察到10倍更高数量的载体基因组和荧光素酶表达。总之，HRII应用程序效率较低（即，较低的荧光素酶活性和载体拷贝数），但对仔猪的痛苦明显较少，并且具有通过内窥镜逆行胰胆管造影术注射（或再注射）载体DNA的潜力。使用病毒载体的肝脏基因治疗是目前最先进的，尽管有副作用的风险。裸DNA载体潜在地避免了这样的风险，但受到低转导效率的限制。在这里，我们表明，直接胆管内注射裸DNA载体的模型系统与新生仔猪是可行的。
Hepatic gene therapy by delivering non-integrating therapeutic vectors in newborns remains challenging due to the risk of dilution and loss of efficacy in the growing liver. Previously we reported on hepatocyte transfection in piglets by intraportal injection of naked DNA vectors. Here, we established delivery of naked DNA vectors to target periportal hepatocytes in weaned pigs by hydrodynamic retrograde intrabiliary injection (HRII). The surgical procedure involved laparotomy and transient isolation of the liver. For vector delivery, a catheter was placed within the common bile duct by enterotomy. Under optimal conditions, no histological abnormalities were observed in liver tissue upon pressurized injections. The transfection of hepatocytes in all tested liver samples was observed with vectors expressing luciferase from a liver-specific promoter. However, vector copy number and luciferase expression were low compared to hydrodynamic intraportal injection. A 10-fold higher number of vector genomes and luciferase expression was observed in pigs using a non-integrating naked DNA vector with the potential for replication. In summary, the HRII application was less efficient (i.e., lower luciferase activity and vector copy numbers) than the intraportal delivery method but was significantly less distressful for the piglets and has the potential for injection (or re-injection) of vector DNA by endoscopic retrograde cholangiopancreatography. Gene therapy for the liver using viral vectors is the current state-of-the-art, despite the risk for side effects. Naked DNA vectors potentially avoid such risks but are limited by low transduction efficacy. Here, we show that direct intrabiliary injection of naked DNA vectors in a model system with newborn piglets is feasible.
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影响因子：
3.7
作者：
Kamimura K;Kanefuji T;Yokoo T;Abe H;Suda T;Kobayashi Y;Zhang G;Aoyagi Y;Liu D
通讯作者：
Liu D
影响因子：
12.4
作者：
Kamimura, Kenya;Suda, Takeshi;Liu, Dexi
通讯作者：
Liu, Dexi
影响因子：
24.5
作者：
Hu, J;Zhang, X;Fabre, JW
通讯作者：
Fabre, JW
影响因子：
7.7
作者：
Adler, DG;Baron, TH;Faigel, DO
通讯作者：
Faigel, DO
DOI：
10.1007/bf02374373
发表时间：
1996-01-01
期刊：
SOMATIC CELL AND MOLECULAR GENETICS
影响因子：
--
作者：
Cabrera, JA;Wilson, JM;Raper, SE
通讯作者：
Raper, SE