Transcriptional Blockades of HIV-1

描述（由申请人提供）：本提案旨在开发HIV-1生命周期的转录阻断。有选择性地操纵控制HIV-1生命周期的基因转录的能力预计将对我们对这种病毒的理解以及它引起的疾病的治疗产生重大影响。这里提出的研究利用了我们设计的转录因子的发展，这些转录因子能够激活或抑制内源性基因。目前还没有其他的基因治疗策略能够同时有效地敲除和上调内源性基因的表达。现在可以制备出具有高亲和力和特异性识别18bp DNA靶序列的多趾锌指蛋白，当融合到激活或抑制结构域时，这些蛋白成为靶基因转录活性的有效调节剂。这一建议的重点是使用我们的转录调节剂特异性地调节HIV-1和宿主基因的转录对其生命周期至关重要。我们的目标是探索靶向基因调节的潜力，作为一种基于基因的治疗策略来治疗HIV-1疾病，以及一种独特的基因发现工具，用于鉴定对病毒至关重要的宿主基因的转录修饰因子。一种全基因组转录调节策略将被开发并应用于寻找新的治疗干预靶点。发现对病毒生命周期至关重要的新型宿主蛋白可能会导致开发一类新的HIV-1药物，因为这些蛋白质可以作为开发传统小分子药物的靶标。有了选择性和有效的转录调节剂，我们将解决设计转录调节剂的潜力，以防止或以其他方式改变原代T细胞和源自CD34+细胞的单核细胞的HIV-1感染过程。这项研究的最终结果应该是增加对HIV-1基因调控的理解，发现新的分子靶点，并开发治疗HIV-1疾病的治疗策略。

DESCRIPTION (provided by applicant): This proposal seeks to develop transcriptional blockades of the HIV-1 lifecycle. The ability to selectively manipulate the transcription of genes controlling the HIV-1 lifecycle is anticipated to have a significant impact on both our understanding of this virus as well as the treatment of the disease it causes. The study proposed here capitalizes on our development of designed transcription factors that enable activation or repression of endogenous genes. Currently no other gene therapy strategy provides the means of both effectively knocking out and up-regulating the expression of an endogenous gene. Polydactyl zinc finger proteins can now be prepared that recognize 18 bp DNA target sequences with high affinity and specificity When fused to activation or repression domains, these proteins become potent regulators of the transcriptional activity of the target gene. This proposal focuses on the use of our transcriptional regulators to specifically modulate transcription of HIV-1 and host genes essential to its lifecycle. We aim to explore the potential of targeted gene modulation as a gene-based therapeutic strategy for the treatment of HIV-1 disease as well as a unique gene discovery tool for the identification of transcriptional modifiers of host genes essential to the virus. A genome-wide transcriptional modulation strategy will be developed and applied to the search for novel targets for therapeutic intervention. The discovery of novel host proteins key to the viral lifecycle could result in the development of a new class of HIV-1 drugs since these proteins could serve as targets for the development of traditional small molecule drugs. With selective and potent transcriptional regulators we will address the potential of designed transcriptional regulators to prevent or otherwise alter the course of HIV-1 infection in primary T cells and monocytes derived from CD34+ cells. The net result of this study should be an increased understanding of HIV-1 gene regulation, the discovery of novel molecular targets, and the development of a therapeutic strategy to treat HIV-1 disease.