Preclinical Development of Ingenol and HDACi Toward HIV Eradication

 描述（由申请人提供）：在接受高效抗逆转录病毒疗法（ART）治疗的个体中存在潜伏性HIV已得到充分证实。在没有额外治疗的情况下，个体必须无限期地保持抗逆转录病毒治疗，以避免严重免疫缺陷的发展和病毒的传播。由于存在这种储存库，治疗中断必然导致病毒反弹。因此，人们普遍认为，根除病毒需要消除潜伏的储存库。 正在研究的一种药物类型是蛋白激酶C激动剂家族，尽管目前还没有进行人体试验。我们和其他人证明，一般来说，PKC激动剂在潜伏期的细胞模型和患者细胞中显示出强烈的活性。Ingenol是一种蛋白激酶C（PKC）激动剂，发现于Euphorbiacea，一个大家庭的肉质植物从半沙漠地区的巴西，已显示出巨大的效力，在离体患者细胞测定和体外系统。 在R21阶段，我们将研究巨大戟醇衍生物如何介导潜伏的HIV的再激活以及PKC亚型的亚群。在目标1中，我们计划研究各种巨大戟醇衍生物的作用机制。在目的2中，我们将研究HDAC抑制剂基于其诱导松弛染色质状态的能力来增强巨大戟醇的活性的作用。这些研究将为目标3提供信息，我们将在目标3中测试 在来自病毒血症患者的细胞中上述药物类别的最佳浓度和组合。这些研究将为在赠款的R33阶段测试这些化合物奠定基础。 R33阶段将包括目标3和4。目标3将检查我们优化的刺激方案对人免疫效应细胞的功能性的潜在影响，无论这些是增强还是可能对其活性有害。被检查的细胞类型将包括自然杀伤细胞、细胞毒性T淋巴细胞、树突状细胞和调节性T细胞。Aim 4将在HIV潜伏期的人源化小鼠模型中测试最有效的巨大戟二萜醇衍生物与HDAC抑制剂的最佳浓度和组合。

 DESCRIPTION (provided by applicant): The presence of latent HIV in individuals treated with highly active antiretroviral therapy (ART) has been well established. Without additional therapies, individuals must remain on ART indefinitely in order to avoid development of severe immunodeficiency and spread of the virus. Because of the existence of this reservoir, treatment interruption invariably leads to viral rebound. Thus, it is generally accepted that eradication of the virus will require elimination of the latent reservoir. A drug type under investigation, although not in human trials at this time, is the family of protein kinase C agonists. We and others demonstrated that PKC agonists, in general, display strong activity across cell models of latency and in patient cells. Ingenol, a protein kinase C (PKC) agonist found in Euphorbiacea, a large family of succulent plants from semi-desertic areas of Brazil, has shown great potency in ex vivo patient cell assays and in in vitro systems. In the R21 phase, we will examine how ingenol derivatives mediate reactivation of latent HIV and which subset(s) of PKC isoforms. In Aim 1, we plan to investigate the mechanism of action of various ingenol derivatives. In Aim 2 we will investigate a role for HDAC inhibitors to enhance the activity of ingenols based on their ability to induce relaxed chromatin states. These studies will inform Aim 3, where we will test the best concentrations and combinations of the above drug categories in cells from aviremic patients. These studies will set the stage for testing of these compounds in the R33 phase of the grant. The R33 phase will consist of Aims 3 and 4. Aim 3 will examine the potential influence that our optimized stimulation regimen(s) will have on the functionality of human immune effector cells, whether these are enhancing or perhaps deleterious to their activities. The cell types being examined will include natural killer cells, cytotoxic T lymphocytes, dendritic cells an regulatory T cells.Aim 4 will test the best concentrations and combinations of the most potent ingenol derivative with and without an HDAC inhibitor in a humanized mouse model of HIV latency.