Inhibition of papilloma progression by antisense oligonucleotides targeted to HPV11 E6/E7 RNA.
Inhibition of papilloma progression by antisense oligonucleotides targeted to HPV11 E6/E7 RNA.
复制标题
通过针对 HPV11 E6/E7 RNA 的反义寡核苷酸抑制乳头状瘤进展。
DOI：
10.1038/sj.gt.3302303
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发表时间：
2004
期刊：
影响因子：
5.1
通讯作者：
Christensen,ND
中科院分区：
文献类型：
--
作者：
Clawson,GA;Miranda,GQ;Sivarajah,A;Xin,P;Pan,W;Thiboutot,D;Christensen,ND
人乳头瘤病毒（HPV）是人类重要的病原体，可引起从良性疣到宫颈发育不良/癌的一系列过度增殖性病变。HPV相关病变需要持续产生致癌E6/E7蛋白，其由双顺反子或重叠mRNA编码。在这里，我们使用分子试剂靶向HPV 11的E6/E7 mRNA，HPV 11是一种与生殖器疣的病因有关的类型。使用文库选择方案鉴定HPV 11 E6/E7 RNA中的可竞争位点，构建靶向这些位点的核酸（DNA酶，反义寡核苷酸），并在细胞培养物和人包皮移植物上进行测试。虽然DNA酶在细胞培养中至少同样有效，但靶向文库选择位点之一（阿索407）周围区域的反义寡核苷酸在阻断免疫缺陷小鼠人包皮移植物中HPV 11诱导的乳头状瘤进展方面最有效。总共有11例乳头状瘤用阿索407治疗。其中，用阿索407治疗的7个小乳头状瘤中的4个通过原位杂交（ISH）显示可检测的病毒丢失，并且在所有这4个中，乳头状瘤在治疗后在大体上或组织学上不再明显。当治疗较大的乳头状瘤时，四分之一的人显示ISH病毒丢失，与乳头状瘤大小的轻微减少有关。考虑到用阿索407治疗的所有11个乳头状瘤，通过ISH的病毒染色的丧失与对照中观察到的显著不同（P< 0.016），对于7个小的经治疗的乳头状瘤也是如此（P< 0.012）。靶向相同位点（或其它文库选择位点）的DNA酶在ISH染色中没有产生统计学显著差异（P< 0.15）。我们对阿索407的研究结果似乎代表了针对真正HPV感染的第一种特异性分子疗法，并为开发靶向其他HPV相关病变的分子疗法提供了合理的原理验证策略。
Human papillomaviruses (HPVs) are recognized as important human pathogens, causing a spectrum of hyperproliferative lesions from benign warts to cervical dysplasias/carcinomas. HPV-associated lesions require continued production of the oncogenic E6/E7 proteins, which are encoded by either bicistronic or overlapping mRNAs. Here we targeted the E6/E7 mRNA of HPV11, a type implicated in causation of genital warts, using molecular reagents. Accessible sites in the HPV11 E6/E7 RNA were identified using library selection protocols, and nucleic acids (DNAzymes, antisense oligonucleotides) targeted to these sites were constructed, and tested in cell culture and on human foreskin grafts. While DNAzymes were at least equally effective in cell culture, antisense oligonucleotides targeted to the region surrounding one of the library-selected sites (ASO 407) proved most effective in blocking progression of HPV11-induced papillomas in human foreskin grafts on immunodeficient mice. In total, 11 papillomas were treated with ASO 407. Of these, four of seven small papillomas treated with ASO 407 showed loss of detectable virus by in situ hybridization (ISH), and in all four of these, papillomas were no longer evident grossly or histologically after treatment. When larger papillomas were treated, one of four showed loss of virus by ISH, associated with a minor decrease in papilloma size. Considering all 11 papillomas treated with ASO 407, loss of viral staining by ISH was significantly different from that observed in controls (P< 0.016), as was true for the seven small treated papillomas (P< 0.012). DNAzymes targeted to the same site (or other library selected sites) did not produce statistically significant differences in ISH staining (P< 0.15). Our results with ASO 407 appear to represent the first specific molecular therapy against a bona fide HPV infection, and provide a rational proof-of-principle strategy for development of molecular therapeutics targeting other HPV-associated lesions.