Smith Theodore A G, Idamakanti Neeraja, Marshall-Neff Jennifer, Rollence Michele L, Wright Patrick, Kaloss Michele, King Laura, Mech Christine, Dinges Lisa, Iverson William O, Sherer Alfred D, Markovits Judit E, Lyons Russette M, Kaleko Michael, Stevenson Susan C
Genetic Therapy, Inc., A Novartis Company, Gaithersburg, MD 20878, USA.
Hum Gene Ther. 2003 Nov 20;14(17):1595-604. doi: 10.1089/104303403322542248.
Adenovirus serotype 5 (Ad5)-based vectors can bind at least three separate cell surface receptors for efficient cell entry: the coxsackie-adenovirus receptor (CAR), alpha nu integrins, and heparan sulfate glycosaminoglycans (HSG). To address the role of each receptor involved in adenoviral cell entry, we mutated critical amino acids in fiber or penton to inhibit receptor interaction. A series of five adenoviral vectors was prepared and the biodistribution of each was previously characterized in mice. To evaluate possible species differences in Ad vector tropism, we characterized the effects of each detargeting mutation in non-human primates after systemic delivery to confirm our conclusions made in mice. In non-human primates, CAR was found to have minimal effects on vector delivery to all organs examined including liver and spleen. Cell-surface alpha nu integrins played a significant role in delivery of vector to the spleen, lung and kidney. The fiber shaft mutation S*, which presumably inhibits HSG binding, was found to significantly decrease delivery to all organs examined. The ability to detarget the liver corresponded with decreased elevations in liver serum enzymes (aspartate transferase [AST] and alanine transferase [ALT]) 24 hr after vector administration and also in serum interleukin (IL)-6 levels 6 hr after vector administration. The biodistribution data generated in cynomolgus monkeys correspond with those data derived from mice, demonstrating that CAR binding is not the major determinant of viral tropism in vivo. Vectors containing the fiber shaft modification may provide for a detargeted adenoviral vector on which to introduce new tropisms for the development of targeted, systemically deliverable adenoviral vectors for human clinical application.
基于5型腺病毒（Ad5）的载体可结合至少三种不同的细胞表面受体以实现高效细胞进入：柯萨奇病毒-腺病毒受体（CAR）、αν整合素和硫酸乙酰肝素糖胺聚糖（HSG）。为了研究参与腺病毒细胞进入的每种受体的作用，我们对纤维或五邻体中的关键氨基酸进行突变以抑制受体相互作用。制备了一系列五种腺病毒载体，并且之前已在小鼠中对每种载体的生物分布进行了表征。为了评估腺病毒载体嗜性中可能存在的物种差异，我们在全身给药后对非人类灵长类动物中每种脱靶突变的作用进行了表征，以证实我们在小鼠中得出的结论。在非人类灵长类动物中，发现CAR对载体递送至所有检查的器官（包括肝脏和脾脏）的影响最小。细胞表面αν整合素在载体递送至脾脏、肺和肾脏中起重要作用。据推测抑制HSG结合的纤维杆突变S*被发现可显著降低载体递送至所有检查器官的效率。使肝脏脱靶的能力与载体给药后24小时肝脏血清酶（天冬氨酸转氨酶[AST]和丙氨酸转氨酶[ALT]）升高的降低以及载体给药后6小时血清白细胞介素（IL）-6水平的降低相对应。食蟹猴中产生的生物分布数据与从小鼠获得的数据一致，表明CAR结合不是体内病毒嗜性的主要决定因素。含有纤维杆修饰的载体可能提供一种脱靶腺病毒载体，在其上可引入新的嗜性，以开发用于人类临床应用的靶向、可全身递送的腺病毒载体。
