Nanoparticles containing siRNA to silence CD4 and CCR5 reduce expression of these receptors and inhibit HIV-1 infection in human female reproductive tract tissue explants.
Nanoparticles containing siRNA to silence CD4 and CCR5 reduce expression of these receptors and inhibit HIV-1 infection in human female reproductive tract tissue explants.
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DOI：
10.4081/idr.2011.e11
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发表时间：
2011-09-07
影响因子：
3.2
通讯作者：
Howell AL
中科院分区：
文献类型：
--
作者：
Eszterhas SK;Ilonzo NO;Crozier JE;Celaj S;Howell AL
人类免疫缺陷病毒1型（HIV-1）与人类女性生殖道靶细胞上的CD 4和CCR 5受体结合。我们试图确定降低女性生殖道细胞中编码这些受体的信使RNA（mRNA）转录物的水平是否可以保护粘膜组织外植体免受HIV-1感染。将从HIV-1血清阴性妇女子宫切除术组织的子宫内膜、子宫颈内膜和子宫颈外膜制备的外植体暴露于含有CD 4和CCR 5特异性短干扰RNA（siRNA）序列的纳米颗粒。两天后将外植体暴露于HIV-1，并在感染后5天测量HIV-1逆转录物。与用不相关的siRNA处理的那些相比，用含有CD 4和CCR 5特异性siRNA的纳米颗粒处理的外植体显示CD 4和CCR 5转录物水平降低，并且HIV-1逆转录物水平显著降低。在雌性生殖道外植体和外周血细胞培养物中，siRNA转染诱导IFN-α（IFN-α）（一种有效的抗病毒细胞因子）的分泌。在雌性小鼠中，子宫内滴注的鼠特异性Cd 4-siRNA纳米颗粒在第3天显著降低了鼠Cd 4转录物。我们的研究结果表明，siRNA纳米颗粒减少人类女性生殖道组织中HIV-1感染性受体的表达，并抑制HIV-1感染。小鼠研究表明，纳米颗粒可以渗透体内生殖道组织并沉默基因表达。siRNA转染后IFN-α的诱导可能有助于抗病毒作用。这些发现支持纳米颗粒的治疗开发，以递送siRNA分子来沉默女性生殖道中的宿主细胞受体，作为抑制粘膜HIV-1传播的新型杀微生物剂。
Human Immunodeficiency Virus-type 1 (HIV-1) binds to CD4 and CCR5 receptors on target cells in the human female reproductive tract. We sought to determine whether reducing levels of messenger RNA (mRNA) transcripts that encode these receptors in female reproductive tract cells could protect mucosal tissue explants from HIV-1 infection. Explants prepared from the endometrium, endocervix, and ectocervix of hysterectomy tissues from HIV-1 sero-negative women were exposed to nanoparticles containing CD4- and CCR5-specific short-interfering RNA (siRNA) sequences. Explants were then exposed two days later to HIV-1, and HIV-1 reverse transcripts were measured five days post-infection. Explants treated with nanoparticles containing CD4- and CCR5-specific siRNA showed reduced levels of CD4 and CCR5 transcripts, and significantly lower levels of HIV-1 reverse transcripts compared to those treated with an irrelevant siRNA. In female reproductive tract explants and in peripheral blood cell cultures, siRNA transfection induced the secretion of IFN-alpha (IFN-α), a potent antiviral cytokine. In female mice, murine-specific Cd4-siRNA nanoparticles instilled within the uterus significantly reduced murine Cd4 transcripts by day 3. Our findings demonstrate that siRNA nanoparticles reduce expression of HIV-1 infectivity receptors in human female reproductive tract tissues and also inhibit HIV-1 infection. Murine studies demonstrate that nanoparticles can penetrate the reproductive tract tissues in vivo and silence gene expression. The induction of IFN-α after siRNA transfection can potentially contribute to the antiviral effect. These findings support the therapeutic development of nanoparticles to deliver siRNA molecules to silence host cell receptors in the female reproductive tract as a novel microbicide to inhibit mucosal HIV-1 transmission.