The kinetics and tissue distribution of protein transduction in mice
The kinetics and tissue distribution of protein transduction in mice
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DOI：
10.1016/j.ejps.2005.10.011
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发表时间：
2006-03-01
影响因子：
4.6
通讯作者：
McLeod, HL
中科院分区：
文献类型：
--
作者：
Cai, SR;Xu, G;McLeod, HL
蛋白质转导域（PTD）为许多疾病的治疗提供了令人兴奋的治疗机会。人类免疫缺陷 1 型 (HIV-1) TAT 蛋白的 11 个氨基酸片段可以将具有生物活性的大蛋白转导至哺乳动物细胞中；最近的证据表明 116 kDa β-半乳糖苷酶蛋白具有体内 PTD。然而，关于 TAT 融合蛋白体内分布的信息很少，无法确定 PTD 在人类研究中的可行性。在这项研究中，我们检查了 PTD 转导的 TAT 融合蛋白在小鼠体内的组织动力学和组织分布。通过四种途径（门静脉、静脉注射、腹腔注射和口服）向小鼠施用低剂量（100μg）或高剂量（500μg）TAT-β-半乳糖苷酶融合蛋白。处理后15分钟、1小时、6小时、10小时和24小时收获组织。通过原位染色、酶活性测定和蛋白质印迹分析来分析β-半乳糖苷酶在各种组织中的分布。在所有组织（肝、肾、脾、肺、肠和脑）中均观察到β-半乳糖苷酶活性。大多数组织中 β-半乳糖苷酶活性在门静脉、静脉注射和腹腔注射后 15 分钟达到峰值。给药后以及口服给药后 1 小时在所有组织中。门静脉注射后 15 分钟，肝脏中的 β-半乳糖苷酶活性 (67 毫单位 [mU]/mg) 高于静脉注射后的水平。 (9.8 mU/mg)，腹腔注射（4.4 mU/mg）和口服（0.3 mU/mg）给药。原位染色和蛋白质印迹结果与β-半乳糖苷酶活性测定密切相关。活性的中位初始半衰期为 2.2 小时，范围为 1.2 小时至 3.4 小时（变异系数 = 28.9%）。口服 PTD 后 β-半乳糖苷酶活性的生物利用度为 24%。这项研究详细介绍了通过 PTD 将 TAT 融合蛋白模型递送至小鼠体内的动力学和组织分布。这些数据允许合理选择治疗性 PTD 的递送途径和时间表，并将有助于在蛋白质疗法的开发中使用 TAT 融合蛋白转导。 (C) 2005 Elsevier B.V. 保留所有权利。
Protein transduction domains (PTDs) offer an exciting therapeutic opportunity for the treatment of many diseases. An 11-amino acid fragment of human immunodeficiency type 1 (HIV-1) TAT-protein can transduce large, biologically active proteins into mammalian cells; recent evidence has shown an in vivo PTD for the 116 kDa beta-galactosidase protein. However, there is little information on the in vivo distribution of the TAT fusion protein to define the viability of PTDs for human studies. In this study we examined the tissue kinetics and tissue distribution of the PTD-transduced TAT fusion protein in mice. Low (100 mu g) or high (500 mu g) doses of TAT-beta-galactosidase fusion protein were administrated to mice through four routes (portal vein, i.v., i.p., and oral). Tissues were harvested 15 min, 1 h, 6 h, 10 h, and 24 h after treatment. Distribution of beta-galactosidase in various tissues was analysed by in situ staining, enzymatic activity assay, and Western blot analysis. p-Galactosidase enzyme activity was observed in all tissues (liver, kidney, spleen, lung, bowel, and brain). beta-Galactosidase activity peaked at 15 min in most tissues after portal vein, i.v., and i.p. administration and at 1 h after oral dosing in all tissues. beta-Galactosidase activity in the liver at 15 min after portal vein injection (67 milliunits [mU]/mg) was higher than after i.v. (9.8 mU/mg), i.p. (4.4 mU/mg), and oral (0.3 mU/mg) dosing. In situ staining and Western blot results correlated closely with beta-galactosidase enzyme activity assay. The median initial half-life for activity was 2.2 h, ranging from 1.2 h to 3.4 h (coefficient of variation = 28.9%). The bioavailability of beta-galactosidase activity after an orally administered PTD was 24%. This study details the kinetics and tissue distribution of delivering of a model TAT fusion protein into the mouse via PTD. These data allow rational selection of delivery route and schedules for therapeutic PTD and will aid the use of TAT fusion protein transduction in the development of protein therapies. (C) 2005 Elsevier B.V. All rights reserved.