Modified adenoviral vectors ablated for coxsackievirus-adenovirus receptor, αv integrin, and heparan sulfate binding reduce in vivo tissue transduction and toxicity
Modified adenoviral vectors ablated for coxsackievirus-adenovirus receptor, αv integrin, and heparan sulfate binding reduce in vivo tissue transduction and toxicity
复制标题
DOI：
10.1089/hum.2006.17.264
复制
发表时间：
2006-03-01
影响因子：
4.2
通讯作者：
Mizuguchi, H
中科院分区：
文献类型：
--
作者：
Koizumi, N;Kawabata, K;Mizuguchi, H
柯萨奇病毒和腺病毒受体（CAR）、α (v)整合素和硫酸肝素糖胺聚糖（hsg）是体内腺病毒（Ad）载体的趋向性决定因素。为了开发有针对性的广告媒介，需要阻止（或减少）其广泛的倾向性。我们之前已经开发了Ad载体，通过纤维突起的FG环突变（删除纤维蛋白的T489， A490， Y491和T492），删除五基的RGD基序，并将纤维轴结构域替换为Ad 35型衍生的结构域，分别切除CAR， α (v)整合素和HSG结合。研究表明，与传统的Ad载体相比，这种三突变Ad载体[Ad/Delta F(FG)Delta P-S35-L2]在小鼠肝脏中的转导明显较低[Koizumi， N., Mizuguchi， H., Sakurai， F., Yamaguchi， T., Watanabe， Y., and Hayakawa， T.(2003)]。[j].中国生物医学工程学报，2016,32(2):444 - 444。在本研究中，我们优化了纤维旋钮突变以进一步减少体内转导，并检测了修饰的Ad载体的毒性。Ad/Delta F(AB)Delta P-S35-L2是一种三突变的Ad载体，含有纤维环中AB环的突变（R412S, A415G， E416G和K417G），通过静脉注射和腹腔注射介导的小鼠肝脏转导分别比传统Ad载体低约15,000和500倍，比Ad/Delta F(FG)Delta P-S35-L2介导的小鼠肝脏转导低10倍。与Ad/ δ F(FG) δ P-S35-L2和传统Ad载体相比，Ad/ δ F(AB) δ P-S35-L2对其他器官的转导也较低。静脉注射Ad/ δ F(AB) δ P-S35-L2后，小鼠血清中肝脏血清酶（天冬氨酸转移酶[AST]和丙氨酸转移酶（ALT））和白细胞介素(IL)-6的水平与未治疗小鼠血清相似，而传统Ad载体导致AST、ALT和IL-6水平升高。因此，我们成功地进一步改进了突变Ad载体，消除了病毒的自然趋向性和毒性。这种新的广告载体似乎是靶向基因传递的基本载体。
Coxsackievirus and adenovirus receptor (CAR), alpha(v) integrins, and heparan sulfate glycosaminoglycans (HSGs) are the tropism determinants of adenoviral (Ad) vectors in vivo. For the development of a targeted Ad vector, its broad tropism needs to be blocked (or reduced). We have previously developed Ad vectors with ablation of CAR, alpha(v) integrin, and HSG binding by mutation of the FG loop in the fiber knob (deletion of T489, A490, Y491, and T492 of the fiber protein), deletion of the RGD motif of the penton base, and substitution of the fiber shaft domain for that derived from Ad type 35, respectively, and have shown that this triple-mutant Ad vector [Ad/Delta F(FG)Delta P-S35-L2] exhibits significantly lower transduction in mouse liver compared with the conventional Ad vector [Koizumi, N., Mizuguchi, H., Sakurai, F., Yamaguchi, T., Watanabe, Y., and Hayakawa, T. ( 2003). J. Virol. 77, 13062-13072]. In the present study, we optimized the fiber knob mutation for further reduced in vivo transduction and examined toxicity of the modified Ad vectors. Ad/Delta F(AB)Delta P-S35-L2, a triple-mutant Ad vector containing a mutation of the AB loop in the fiber knob (R412S, A415G, E416G, and K417G), mediated approximately 15,000- and 500-fold lower mouse liver transduction by intravenous and intraperitoneal administration, respectively, than the conventional Ad vector, and mediated 10-fold lower mouse liver transduction than did Ad/Delta F(FG)Delta P-S35-L2. Ad/Delta F(AB)Delta P-S35-L2 also exhibited lower transduction of other organs compared with Ad/Delta F(FG)Delta P-S35-L2 and the conventional Ad vector. Levels of both liver serum enzymes (aspartate transferase [AST] and alanine transferase (ALT)] and interleukin (IL)-6 in mouse serum after intravenous administration of Ad/Delta F(AB)Delta P-S35-L2 were similar to those in the nontreatment mouse serum, whereas the conventional Ad vector led to high levels of AST, ALT, and IL-6. We therefore succeeded in further improving the mutant Ad vector, abolishing both viral natural tropism and toxicity. This new Ad vector appears to be a fundamental vector for targeted gene delivery.