ANTIVIRAL DRUG RESISTANCE IN HUMAN CYTOMEGALOVIRUS

描述：(改编自申请者摘要) 治疗艾滋病患者巨细胞病毒感染的方法是 它需要长期的治疗，通过这种治疗，有 耐药病毒的出现和疾病的进展。目前， CMV的耐药性监测受到经典细胞的限制。 养殖技术效率低，不能及时提供 反馈。这个项目的目的是研究基因的变化。 出现在耐药的临床巨细胞病毒分离株中，作为一种手段 快速诊断并作为对该病生物学特性的指导 抗药性病毒。人们的预期是，一组有限的突变 在CMV的两个关键基因中，UL97磷酸转移酶(UL97)和DNA中 聚合酶(Poll)，将一直与耐药有关 病毒，在治疗过程中耐药突变的进化 与治疗失败相关，并导致特定的突变 对多种药物产生交叉耐药性或酶活性改变 这可能会影响病毒的复制效率，并最终影响毒力。 该项目的具体目标是：1)确定 UL97和Poll在临床上对抗CMV药物的耐药性 使用，例如更昔洛韦、磷卡奈特、西多福韦、洛布卡韦和阿昔洛韦， 2)跟踪连续分离株中耐药突变的演变 来自接受治疗的患者与药物方案的关系，以及临床 和病毒学结果；以及3)检查病毒的生物学特性 抗药性CMV突变体的交叉耐药性、复制 能力、遗传稳定性和磷酸转移酶和 聚合酶活性。在这方面，单一和多重的影响 我们将检查突变情况。这项工作将在 孙雯在俄勒冈健康科学大学的实验室和 将利用W.博士实验室提供的临床分离株。 劳伦斯·德鲁(加州大学旧金山分校)和阿莱霍·埃赖斯博士(明尼苏达大学)。

DESCRIPTION: (Adapted from Applicant's Abstract) The problem with treatment of cytomegalovirus (CMV) infection in AIDS patients is that it requires long- term treatment, and, with this treatment, there is emergence of drug resistant virus and disease progression. At present, the monitoring of drug resistance in CMV is limited by classical cell culture techniques which are inefficient and do not provide timely feedback. The purpose of this project is to study the genetic changes that occur in drug-resistant clinical CMV isolates, both as a means of rapid diagnosis and as a quide to the biological characteristics of the resistant viruses. The expectation is that a limited set of mutations in two key CMV genes, the UL97 phosphotransferase (UL97) and in the DNA polymerase (pol), will consistently be associated with drug-resistant viruses, that the evolution of resistance mutations during therapy is associated with treatment failure, and that specific mutations result in cross-resistance to multiple drugs or in altered enzyme activity which may effect viral replication efficiency and, ultimately, virulence. The specific aims of this project are 1) to identify the mutations in UL97 and pol which can confer resistance to anti-CMV drugs in clinical use, e.g., ganciclovir, foscarnet, cidofovir, lobucavir, and acyclovir, 2) to track the evolution of resistance mutations in serial isolates from treated patients in relation to drug regimen, as well as clinical and virological outcome; and 3) to examine the biological properties of drug resistant CMV mutants with respect to cross- resistance, replication competence, genetic stability and alterations in phosphotransferase and polymerase activity. In this regard, the effects of single and multiple mutations will be examined. The work will be performed in the laboratory of Sun Wen Chu at the Oregon Health Sciences University and will utilize clinical isolates provided from the laboratory of Dr. W. Lawrence Drew (UCSF) and from Dr. Alejo Erice (University of Minnesota).