Safety evaluation of hemagglutinating virus of Japan-artificial viral envelope liposomes in nonhuman primates
Safety evaluation of hemagglutinating virus of Japan-artificial viral envelope liposomes in nonhuman primates
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DOI：
10.1089/104303401300042366
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发表时间：
2001-03-20
影响因子：
4.2
通讯作者：
Kaneda, Y
中科院分区：
文献类型：
--
作者：
Tsuboniwa, N;Morishita, R;Kaneda, Y
通过一系列实验，我们在食蟹猴身上测试了一种混合脂质体载体--日本血凝病毒(HVJ)-人工病毒被膜(AVE)脂质体用于人类治疗性基因转移的安全性和有效性。在一项重复的肌肉给药研究中，使用HVJ-AVE脂质体混悬液治疗的赋形剂对照猕猴(n=2)，接受了重复肌肉注射2毫升的试验物质。人肝细胞生长因子(HGF)c DNA插入表达载体(PUC-SRα/HGF)注射动物(n=2)，注射含PUC-SRα/HGF的HVJ-AVE脂质体混悬液，重复肌肉注射受试物2mI。分别于治疗前、治疗后7、21、28、29天进行一般身体状况、血液学、血液化学、血清HGF检测。注射PUC-SRα/HGF的猕猴HGF水平升高。观察结束后，处死猕猴进行尸检和组织学检查，用聚合酶链式反应(PCR)分析肝、脾和注射部位的PUC-SRα/HGF。在单次静脉给药研究中，对照猕猴(n=4)接受单次静脉注射10ml生理盐水。赋形剂对照组动物(n=5)单次静脉注射10mlHVJ-AVE脂质体混悬液。DNA处理组(n=7)一次静脉注射含有DNA[pcDNA3.1(+)]的HVJ-AVE脂质体混悬液10ml。分别于治疗前、治疗后第1、14、21、28天检测一般情况、体重、血液学、血生化、尿液成分。观察结束后，处死猕猴进行尸检和组织学检查。第1天所有猕猴的肺、肝、脾、肾和心脏组织中均检测到pcDNA3.1(+)。然而，在第14天、第21天和第28天检查的任何组织中都没有检测到DNA。反转录(RT)-聚合酶链式反应(RT-PCR)未检测到病毒基因组RNA。在这一系列的安全评估中，动物在没有改变体重或一般情况的情况下耐受了安全研究。未检测到血液学改变或血液化学或尿液成分的改变。此外，未观察到组织学变化。这项安全性评价研究证明了新型转基因载体HVJ-AVE脂质体在人体内的安全性、可行性和治疗潜力。
We tested, in cynomolgus monkeys, the safety and effectiveness of a hybrid liposome vector, hemagglutinating virus of Japan (HVJ)-artificial viral envelope (AVE) liposomes, for human therapeutic gene transfer in a series of experiments. In a repetitive intramuscular administration study, vehicle control macaques (n = 2), which were treated with HVJ-AVE liposome suspension, received repetitive intramuscular injections of 2 ml of test substance. Human hepatocyte growth factor (HGF) cDNA-inserted expression vector (PUC-SR alpha /HGF) injection animals (n = 2), which were treated with HVJ-AVE liposome suspension containing pUC-SR alpha /HGF, received repetitive intramuscular injection of 2 mi of test substance. General body condition, hematology, blood chemistry, and serum HGF were determined sequentially before treatment and 7, 21, 28, and 29 days after treatment. Elevations in HGF were detected in monkeys injected with pUC-SR alpha /HGF. After this observation period, macaques were killed for autopsy and histological examination, pUC-SR alpha /HGF was detected by polymerase chain reaction (PCR) analysis in the liver, spleen, and at the injection site. In single intravenous administration study, control macaques (n = 4) received a single intravenous injection of 10 ml of physiological saline. Vehicle control animals (n = 5) received a single intravenous injection of 10 ml of HVJ-AVE liposome suspension. DNA-treated animals (n = 7) received a single intravenous injection of 10 ml of HVJ-AVE liposome suspension containing plasmid DNA [pcDNA 3.1(+)]. General body condition, body weight, hematology, blood chemistry, and urine composition were determined sequentially before treatment and 1, 14, 21, and 28 days after treatment. After this observation period, macaques were killed for autopsy and histological examination. pcDNA 3.1(+) was detected by PCR analysis on day 1 in lung, liver, and spleen of all monkeys, in kidney of one of two monkeys, and in heart of one of two monkeys. However, no DNA was detected in any of the tissues examined on days 14, 21, and 28. No virus genomic RNA was detected by reverse transcription (RT)-PCR analysis with HVJ-specific primers. In this series of safety evaluations, the animal tolerated the safety study with no change in body weight or general condition. No hematological changes or alterations in blood chemistry or urine composition was detected. Moreover, no histological changes were observed. This safety evaluation study demonstrates the safety, feasibility, and therapeutic potential of the novel transfection vehicle, HVJ-AVE liposomes, in humans.