Novel Anti-HCMV Strategies

项目总结 人类巨细胞病毒(HCMV)是导致移植失败的最重要的单一感染和 仍然是导致大量发病和死亡的一个原因。事实上，巨细胞病毒病发生在20%-30%的 移植有感染的风险，对于肺或心肺移植受者来说是一个特别的问题，据报道 发病率为50-80%。巨细胞病毒的临床表现具有广泛性、炎症性和依赖性。 关于终末器官功能障碍。与巨细胞病毒感染相关的炎症性器官疾病是一种直接的 全身性病毒传播到多个器官部位并感染的后果 无症状或有症状的感染。单核细胞负责将病毒传递到组织和 在感染器官的炎症状态中起着核心作用。由于针对巨细胞病毒的治疗旨在 阻断病毒复制周期中的特定步骤，在感染的血液单核细胞中缺乏HCMV复制 表明HCMV抗病毒药物在预防病毒的初始传播方面并不有效。一致同意， 预防措施只是将人巨细胞病毒病的动力学转移到移植后的较晚时间，因为无法 抗病毒药物消除浸润性感染的炎性髓系细胞，这是主要的细胞类型 在移植患者被感染的器官中发现。因此，我们主张抑制人巨细胞病毒复制 预防性抗病毒药物必须与新药联合使用，特别是 杀死受感染的单核细胞。我们发现巨细胞病毒明确地利用细胞内的Mcl-1，一种抗细胞凋亡剂 Bcl2蛋白家族的成员，刺激受感染的单核细胞存活。使用高- 通过吞吐量筛选方法，Nikolovska-Coleska博士的实验室合成并表征了两种 新型高选择性Mcl-1拮抗剂。我们测试了每一类化合物中的一种先导化合物 两者对HCMV感染的单核细胞均具有较高的杀伤活性。此外， 我们能够显著提高每种化合物的选择性和杀灭活性 纳米粒子是在罗博士的实验室里开发出来的。因此，我们假设Mcl-1抑制剂将针对HCMV-1。 感染单核细胞以防止血源性传播，这将在3个不同的目标中进行测试。目标1 将进一步评估Mcl-1小分子抑制剂的两个库类，以确定显示出 对巨细胞病毒感染的单核细胞的最大细胞毒作用。药物化学也将预习到 提高化合物的效力。目标2将是进一步开发和表征纳米颗粒技术 以增强Mcl-1抑制剂对感染单核细胞的靶向性。目标3将确定FREE的疗效 与微囊化Mcl-1小分子抑制剂中和/限制人巨细胞病毒人源化传播的比较 老鼠模型。这些研究将评估Mcl-1抑制剂作为针对感染者的抗病毒策略的使用。 细胞而不是直接感染病毒，这可能会对移植患者的预后产生重大影响 人巨细胞病毒感染的高危人群。

PROJECT SUMMARY Human cytomegalovirus (HCMV) is the single most important infection leading to transplant failure and continues to be a cause of substantial morbidity and death. Indeed, HCMV disease occurs in 20-30% of transplants at risk for infection and is a particular problem to lung or heart-lung recipients with a reported incidence of 50-80%. Clinical manifestations of HCMV are widespread, inflammatory in nature, and dependent on end-organ dysfunction. Inflammatory organ diseases associated with a HCMV infection is a direct consequence of the systemic viral spread to and infection of multiple organ sites that occur during either asymptomatic or symptomatic infections. Monocytes are responsible for delivering the virus into tissue and play a central role in the inflammatory state of infected organs. Since therapies against HCMV are designed to block specific steps along the virus replication cycle, the lack of HCMV replication in infected blood monocytes indicates that HCMV antiviral drugs are not effective in preventing the initial spread of the virus. In accord, prophylaxis has simply shifted the kinetics of HCMV disease to later after transplantation due to the inability of antiviral drugs to eliminate infiltrating infected inflammatory myeloid cells, which are the principle cell type found in infected organs of transplant patients. Thus, we advocate that the suppression of HCMV replication with prophylactic antiviral drugs must be administered in combination with novel drugs specifically capable of killing infected monocytes. We have found that HCMV explicitly utilizes cellular Mcl-1, an antiapoptotic member of the Bcl-2 family of proteins, to stimulate the survival of infected monocytes. Using a high- throughput screening approach, Dr. Nikolovska-Coleska’s laboratory has synthesized and characterized two novel classes of highly selective Mcl-1 antagonists. We have tested one lead compound from each class of inhibitor and demonstrated both to have high killing activity towards HCMV-infected monocytes. Furthermore, we were able to significantly enhance the selectivity and killing activity of each compound by encapsulation into nanoparticles developed in Dr. Luo’s laboratory. Thus, we hypothesize that Mcl-1 inhibitors will target HCMV- infected monocytes to prevent hematogenous dissemination, which will be tested in 3 separate aims. Aim 1 will further evaluate the two library classes of Mcl-1 small-molecule inhibitors to identify compounds exhibiting maximum cytotoxicity against HCMV-infected monocytes. Medicinal chemistry will also be preformed to increase the potency of compounds. Aim 2 will be to further develop and characterize nanoparticle technology to enhance targeting of Mcl-1 inhibitors towards infected monocytes. Aim 3 will determine the efficacy of free versus encapsulated Mcl-1 small-molecule inhibitors at neutralizing/limiting HCMV spread in a humanized mouse model. These studies will evaluate the use of Mcl-1 inhibitors as an antiviral strategy to target infected cells rather than the virus directly, which could have major impact on the prognosis of transplant patients at high-risk for HCMV infection.