Inhibitors of Kaposi???s Sarcoma Herpesvirus

描述(申请人提供)：卡波西肉瘤相关疱疹病毒(KSHV)在卡波西肉瘤和原发渗出性淋巴瘤中起病因学作用。KSHV潜伏感染肿瘤细胞。在潜伏感染中，病毒基因组以多拷贝、染色体外质粒(Episome)的形式存在于肿瘤细胞中。为了维持下去，病毒基因组必须在每一次细胞分裂时复制，然后分离到子细胞核。KSHV潜伏期相关核抗原(LANA)确保了这些任务的执行，并确保了病毒DNA有效地转移到后代肿瘤细胞。LANA将KSHV DNA拴在有丝分裂染色体上，使病毒附体有效地分离到子细胞核。C-末端LANA自结合结合KSHV末端重复(TR)元件中的特定DNA序列。这种C-末端LANA DNA结合对于KSHV DNA复制、与有丝分裂染色体的连接和Episome持久性是必不可少的。目前，还没有LANA的小分子抑制剂。由于LANA是KSHV潜伏感染的关键，阻断LANA与TRDNA结合的化学探针将成为研究LANA和KSHV生物学的非常有用的试剂。由于肿瘤细胞的持久性依赖于KSHV感染，这种抑制性小分子将具有潜在的治疗益处。这项工作将开发一种强大的、可重复的和小型化的荧光偏振(FP)分析方法，用于高通量筛选抑制KSHV LANA与其DNA识别序列结合的小分子。使用FP检测的中试筛查将在高通量筛查之前验证该检测。 公共卫生相关性：卡波西肉瘤相关疱疹病毒在某些人类癌症中起致病作用。这项工作将开发测试，以确定可用于研究这种病毒的化合物。这些化合物可能被用来预防和治疗这种病毒引起的癌症。

DESCRIPTION (provided by applicant): Kaposi's sarcoma-associated herpesvirus (KSHV) has an etiologic role in Kaposi's sarcoma and primary effusion lymphoma. KSHV latently infects tumor cells. In latent infection, the viral genome persists in tumor cells as a multiple copy, extrachromosomal plasmid (episome). To persist, the viral genome must replicate with each cell division and then segregate to daughter cell nuclei. KSHV latency-associated nuclear antigen (LANA) ensures these tasks are performed and that efficient transfer of the viral DNA to progeny tumor cells occurs. LANA tethers KSHV DNA to mitotic chromosomes to allow efficient segregation of viral episomes to daughter cell nuclei. C-terminal LANA self-associates to bind specific DNA sequence in the KSHV terminal repeat (TR) elements. This C-terminal LANA DNA binding is essential for KSHV DNA replication, tethering to mitotic chromosomes, and episome persistence. Currently, there are no small molecule inhibitors for LANA. Since LANA is critical for KSHV latent infection, chemical probes which block the essential LANA binding to the TR DNA would serve as extremely useful reagents to investigate LANA and KSHV biology. Such inhibitory small molecules would be of potential therapeutic benefit since tumor cell persistence is dependent on KSHV infection. This work will develop a robust, reproducible, and miniaturized fluorescence polarization (FP) assay for a high throughput screen for small molecules that inhibit KSHV LANA binding to its DNA recognition sequence. Pilot screens using the FP assay will validate the assay prior to high throughput screening. PUBLIC HEALTH RELEVANCE: Kaposi's sarcoma-associated herpesvirus has a causative role in certain human cancers. This work will develop tests to identify compounds that can be used to study this virus. These compounds may potentially be used to prevent and treat the cancers this virus causes.