Cancer Gene Therapy Group, Molecular Cancer Biology Program and Transplantation Laboratory, Haartman Institute, University of Helsinki, Finland.
J Gene Med. 2009 Nov;11(11):966-77. doi: 10.1002/jgm.1373.
Rapid clearance of adenoviruses from blood by macrophage lineage cells of the liver and spleen, and binding to platelets, hinder their successful systemic use for cancer gene therapy. Vitamin K dependent coagulation factors are important mediators for the adenovirus liver tropism. Here we aim to determine the effects of coagulation factor, thrombocyte and liver macrophage (Kupffer cell) ablation on biodistribution of serotype 5 adenoviruses in mice with orthotopic breast tumors.
Prior to intravenous injection of adenoviruses, mice bearing orthotopic breast tumors were pretreated with warfarin to inhibit vitamin K dependent coagulation factor synthesis, an anti-platelet antibody causing thrombocytopenia or an inhibitor of the Kupffer cell scavenger receptor or their combination. Virus availability in blood after injection was determined from blood samples and transgene expression in tissues analyzed 72 hours afterwards with In Vivo Imaging and luciferase assays.
Warfarin pretreatment reduced gene delivery to liver, spleen and lung. Kupffer cell ablation increased persistence of adenoviruses in blood but didn't affect biodistribution significantly. Depletion of Kupffer cells combined with thrombocytopenia doubled the systemic gene delivery of 5/3 chimeric adenovirus to tumors (p < 0.05). Triple ablation of platelets, Kupffer cells and coagulation factors increased the tumor to liver ratio of systemic adenovirus gene delivery by 81% (p < 0.05).
Depletion of coagulation factors can reduce transduction of liver, spleen and lung. Kupffer cell depletion is the most feasible method of increasing amount adenovirus in systemic blood flow and in combination with ablation of thrombocytes can increase the transduction of adenovirus to tumors.
肝和脾中的巨噬细胞谱系细胞以及血小板对腺病毒的快速清除和结合，阻碍了其成功用于癌症基因治疗的全身性应用。维生素 K 依赖性凝血因子是腺病毒肝脏趋向性的重要介质。在这里，我们旨在确定凝血因子、血小板和肝巨噬细胞（枯否细胞）耗竭对原位乳腺癌小鼠中血清型 5 腺病毒生物分布的影响。
在静脉注射腺病毒之前，用华法林预处理携带原位乳腺癌的小鼠，以抑制维生素 K 依赖性凝血因子合成、导致血小板减少的抗血小板抗体或枯否细胞清除受体抑制剂或它们的组合。注射后从血液样本中确定病毒在血液中的可用性，并在 72 小时后通过体内成像和荧光素酶测定分析组织中转基因表达。
华法林预处理降低了肝脏、脾脏和肺部的基因传递。枯否细胞耗竭增加了腺病毒在血液中的持续时间，但对生物分布没有显著影响。枯否细胞耗竭与血小板耗竭相结合使 5/3 嵌合腺病毒的全身基因传递增加了一倍（p<0.05）。血小板、枯否细胞和凝血因子的三重耗竭使全身腺病毒基因传递到肿瘤的肿瘤与肝脏比值增加了 81%（p<0.05）。
凝血因子的耗竭可以减少肝脏、脾脏和肺部的转导。枯否细胞耗竭是增加全身血流中腺病毒数量的最可行方法，与血小板耗竭相结合可以增加腺病毒对肿瘤的转导。
