Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice.
Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice.
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DOI：
10.1016/j.omtm.2022.10.006
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发表时间：
2022-12-08
期刊：
影响因子：
--
通讯作者：
Grisch-Chan, Hiu Man
中科院分区：
文献类型：
--
作者：
Deplazes, Sereina;Schlegel, Andrea;Song, Zhuolun;Allegri, Gabriella;Rimann, Nicole;Scherer, Tanja;Willimann, Melanie;Opitz, Lennart;Cunningham, Sharon C.;Alexander, Ian E.;Kipar, Anja;Haeberle, Johannes;Thoeny, Beat;Grisch-Chan, Hiu Man
流体动力学尾静脉注射（HTV）是用于将裸DNA载体递送至小鼠肝脏的“金标准”，从而主要去除静脉周肝细胞。虽然HTV可以纠正代谢性肝脏缺陷，如苯丙酮尿症或胱硫醚β-合酶缺乏症，但尽管在肝脏中过表达，但纠正具有鸟氨酸转氨甲酰酶（OTC）缺乏症的spfash小鼠是不可能的，因为OTC酶主要在门静脉周围肝细胞中表达。为了靶向门静脉周围肝细胞，我们在小鼠中建立了流体动力学逆行胆管内注射（HRII），并使用荧光素酶作为标记基因优化了微环（MC）载体递送。HRII导致转染效率低于1%，比HTV低100倍。虽然HRII诱导最小的肝毒性相比，HTV，荧光素酶的过表达，这两种方法，但不是一个天然的肝脏特异性酶，引起免疫反应，导致消除荧光素酶的表达。在spfash小鼠中通过HRII递送的MC载体的进一步测试没有产生足够的治疗功效，并且需要进一步优化和/或选择校正的细胞。这项研究表明，荧光素酶的表达对肝脏是有毒的。此外，通过胆管的MC载体的物理递送具有治疗局限于门静脉周围肝细胞的缺陷的潜力，这为非病毒肝脏定向基因治疗打开了新的大门。流体动力学尾静脉注射是一种通过主要切除静脉周围肝细胞将DNA递送到小鼠肝脏的工具。由于一些（代谢）途径位于门静脉周围组织中，我们使用荧光素酶作为标记物建立了通过胆管的（逆行）输注，并发现荧光素酶的过表达具有肝毒性。
Hydrodynamic tail vein injection (HTV) is the “gold standard” for delivering naked DNA vectors to mouse liver, thereby transfecting predominately perivenous hepatocytes. While HTV corrects metabolic liver defects such as phenylketonuria or cystathionine β-synthase deficiency, correction of spfash mice with ornithine transcarbamylase (OTC) deficiency was not possible despite overexpression in the liver, as the OTC enzyme is primarily expressed in periportal hepatocytes. To target periportal hepatocytes, we established hydrodynamic retrograde intrabiliary injection (HRII) in mice and optimized minicircle (MC) vector delivery using luciferase as a marker gene. HRII resulted in a transfection efficiency below 1%, 100-fold lower than HTV. While HRII induced minimal liver toxicity compared with HTV, overexpression of luciferase by both methods, but not of a natural liver-specific enzyme, elicited an immune response that led to the elimination of luciferase expression. Further testing of MC vectors delivered via HRII in spfash mice did not result in sufficient therapeutic efficacy and needs further optimization and/or selection of the corrected cells. This study reveals that luciferase expression is toxic for the liver. Furthermore, physical delivery of MC vectors via the bile duct has the potential to treat defects restricted to periportal hepatocytes, which opens new doors for non-viral liver-directed gene therapy. Hydrodynamic tail vein injection is a tool to deliver DNA to murine liver by transfecting primarily perivenous hepatocytes. As some (metabolic) pathways are located in the periportal tissue, we established (retrograde) infusion via the bile duct using luciferase as a marker and found that overexpression of luciferase is liver toxic.
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影响因子：
12.4
作者：
Cunningham, Sharon C.;Kok, Cindy Y.;Alexander, Ian E.
通讯作者：
Alexander, Ian E.
DOI：
10.1083/jcb.105.6.2631
发表时间：
1987-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Sztul ES;Hendrick JP;Kraus JP;Wall D;Kalousek F;Rosenberg LE
通讯作者：
Rosenberg LE
影响因子：
4.6
作者：
Forootan SS;Mutter FE;Kipar A;Iwawaki T;Francis B;Goldring CE;Park BK;Copple IM
通讯作者：
Copple IM
DOI：
10.1016/j.omtm.2022.01.006
发表时间：
2022-03-10
期刊：
Molecular therapy. Methods & clinical development
影响因子：
--
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
通讯作者：
Thöny B
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