Inhibitors of Kaposi???s Sarcoma Herpesvirus

描述（由申请方提供）：卡波西肉瘤相关疱疹病毒（KSHV）在卡波西肉瘤和原发性渗出性淋巴瘤中具有病原学作用。KSHV潜伏感染肿瘤细胞。在潜伏感染中，病毒基因组作为多拷贝的染色体外质粒（附加体）存在于肿瘤细胞中。为了持续存在，病毒基因组必须随着每次细胞分裂而复制，然后分离到子细胞核中。KSHV潜伏相关核抗原（拉娜）确保这些任务得以执行，并确保病毒DNA有效转移至子代肿瘤细胞。拉娜将KSHV DNA连接到有丝分裂染色体上，以使病毒附加体有效分离到子细胞核中。C-末端拉娜自缔合以结合KSHV末端重复（TR）元件中的特异性DNA序列。这种C-末端拉娜DNA结合对于KSHV DNA复制、与有丝分裂染色体的束缚和附加体持久性是必需的。目前，没有拉娜的小分子抑制剂。由于拉娜是KSHV潜伏感染的关键，阻断必需的拉娜与TR DNA结合的化学探针将作为研究拉娜和KSHV生物学的非常有用的试剂。这种抑制性小分子将具有潜在的治疗益处，因为肿瘤细胞的持久性取决于KSHV感染。这项工作将开发一个强大的，可重复的，和小型化的荧光偏振（FP）检测高通量筛选抑制KSHV拉娜结合其DNA识别序列的小分子。在高通量筛选之前，使用FP检测试剂盒进行中试筛选将验证该检测试剂盒。 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具有致病作用。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。这些化合物有可能用于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.