Molecular Analysis of HIV-1C Transmission Cofactors

描述（由申请人提供）：尽管现在许多努力都集中在预防HIV-1在非洲的传播上，但关于决定传播结果的潜在分子机制的信息仍然很少。虽然众所周知，母亲艾滋病毒血浆RNA越高，母婴传播的风险就越高，但没有确定一个病毒阈值，超过这个阈值就绝对传播，低于这个阈值就不会发生传播。因此，我们假设HIV-1的传播受到一系列复杂的宿主细胞辅助因子的影响，这些辅助因子通过影响局部病毒的进入和复制来促进（或保护）病毒传播。在这个新的研究者修订中，我们建议评估HIV-1传播的分子相关因素，我们称之为“co, actors”，以更好地定义围产期传播的决定因素。我们将利用一个优秀的标本库，该样本库已建立，用于从博茨瓦纳未使用药物的HIV-1C阳性母亲及其婴儿中横断面获得的传播液，通过母体标本的RNA表达谱来识别与围产期传播相关的传播辅助因子。然后，我们将使用实时PCR验证辅因子水平和病毒表达水平。最后，我们将利用HIV-1C和HIV-1B的全基因组分子克隆，在体外评估已鉴定的传播辅助因子对HIV-1复制的影响。这些目标旨在确定和功能表征分子辅助因子对HIV-1进入和复制的影响，以便更好地确定传播的生物学决定因素。该提案的一个关键特征是旨在促进波士顿大学医学院（Boston， MA）、波士顿大学公共卫生学院和博茨瓦纳-哈佛伙伴关系之间的合作。这种合作努力对于阐明HIV-1C传播的分子机制至关重要，应被视为研究重点。

DESCRIPTION (provided by applicant): Although many efforts are now focused on preventing HIV-1 transmission within Africa, there remains very little information regarding the underlying molecular mechanisms that determine transmission outcome. While it is well established that the risk of mother-to-child-transmission increases with higher maternal HIV plasma RNA, a threshold of virus has not been identified above which transmission is absolute, nor has a threshold been identified below which transmission does not occur. We therefore hypothesize that HIV-1 transmission is influenced by a complex array of host cellular cofactors that differentially promote (or protect from) viral transmission by influencing local viral entry and replication. In this new investigator revision, we propose to evaluate molecular correlates of HIV-1 transmission, which we term, "co, actors", to better define determinants for perinatal transmission. We will utilize an excellent specimen bank that has been established for transmission fluids obtained cross-sectionally from drug naive, HIV-1C positive mothers and their infants in Botswana to identify transmission cofactors through RNA expression profiling of maternal specimens in association with perinatal transmission. We will then validate cofactor levels and viral expression levels using real-time PCR. Finally, we will evaluate the influence of identified transmission cofactors on HIV-1 replication, in vitro, using full genome molecular clones for HIV-1C and HIV-1B. These aims are designed to identify and functionally characterize the influence of molecular cofactors on HIV-1 entry and replication, to better define biological determinants for transmission. A key feature of this proposal is the intent to foster collaboration between the Boston University School of Medicine (Boston, MA), the Boston University School of Public Health and the Botswana-Harvard Partnership This collaborative effort is essential to elucidate the molecular mechanisms involved in HIV-1C transmission and should be considered a research priority.