Cytotoxicological Analysis of a gp120 Binding Aptamer with Cross-Clade Human Immunodeficiency Virus Type 1 Entry Inhibition Properties: Comparison to Conventional Antiretrovirals
Cytotoxicological Analysis of a gp120 Binding Aptamer with Cross-Clade Human Immunodeficiency Virus Type 1 Entry Inhibition Properties: Comparison to Conventional Antiretrovirals
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DOI：
10.1128/aac.01502-08
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发表时间：
2009-07-01
影响因子：
4.9
通讯作者：
Khati, Makobetsa
中科院分区：
文献类型：
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作者：
de Campos, Walter Rangel Lopes;Coopusamy, Dayaneethie;Khati, Makobetsa
抗逆转录病毒药物（ARV）的长期累积细胞毒性是人类免疫缺陷病毒（HIV）感染者和艾滋病患者治疗失败的主要原因之一。这就需要开发细胞毒性较小或无细胞毒性的新型抗逆转录病毒药物。在本研究中，我们比较了一种称为 B40 的跨进化枝 HIV 1 型中和适体与一组非核苷逆转录酶抑制剂 (NNRTI)、核苷逆转录酶抑制剂 (NRTI)、蛋白酶抑制剂 (PI) 和进入抑制剂 (EI) T20 对人心肌细胞和外周血单核细胞的细胞毒性作用。使用细胞死亡作为终点测量的初始筛选显示，B40 适体和 T20 是唯一在最大使用浓度下对两种细胞类型的活力没有显着影响 (0.61 < P < 0.92) 的测试分子。 PI 类毒性最强 (0.001 < P < 0.00001)，其次是 NNRTI 和 NRTI (0.1 < P < 0.00001)。进一线粒体引发的细胞毒性与抗逆转录病毒药物的使用密切相关。因此，我们使用实时 PCR 来量化线粒体 DNA 与核 DNA 的相对比率，作为毒性标记。与未处理的对照细胞中的水平相比，暴露于 B40 适体的细胞中的线粒体 DNA 水平保持不变 (0.5 > P > 0.06)。这些数据支持将 B40 和相关 EI 适体开发为新的抗逆转录病毒药物，在估计的潜在治疗剂量下没有细胞毒性。
The long-term cumulative cytotoxicity of antiretrovirals (ARVs) is among the major causes of treatment failure in patients infected with human immunodeficiency virus (HIV) and patients with AIDS. This calls for the development of novel ARVs with less or no cytotoxicity. In the present study, we compared the cytotoxic effects of a cross-clade HIV type 1-neutralizing aptamer called B40 with those of a panel of nonnucleoside reverse transcriptase inhibitors (NNRTIs), nucleoside reverse transcriptase inhibitors (NRTIs), protease inhibitors (PIs), and the entry inhibitor (EI) T20 in human cardiomyocytes and peripheral blood mononuclear cells. An initial screen in which cell death was used as the end-point measurement revealed that the B40 aptamer and T20 were the only test molecules that had insignificant (0.61 < P < 0.92) effects on the viability of both cell types at the maximum concentration used. PIs were the most toxic class (0.001 < P < 0.00001), followed by NNRTIs and NRTIs (0.1 < P < 0.00001). Further studies revealed that B40 and T20 did not interfere with the cellular activity of the cytochrome P450 3A4 enzyme (0.78 < P < 0.24) or monoamine oxidases A and B (0.83 < P < 0.56) when the activities of the enzymes were compared to those in untreated controls of both cell types. Mitochondrion-initiated cellular toxicity is closely associated with the use of ARVs. Therefore, we used real-time PCR to quantify the relative ratio of mitochondrial DNA to nuclear DNA as a marker of toxicity. The levels of mitochondrial DNA remained unchanged in cells exposed to the B40 aptamer compared to the levels in untreated control cells (0.5 > P > 0.06). These data support the development of B40 and related EI aptamers as new ARVs with no cytotoxicity at the estimated potential therapeutic dose.