Identification of new antiviral agents against Kaposi's sarcoma-associated herpesvirus (KSHV) by high-throughput drug screening reveals the role of histamine-related signaling in promoting viral lytic reactivation
Identification of new antiviral agents against Kaposi's sarcoma-associated herpesvirus (KSHV) by high-throughput drug screening reveals the role of histamine-related signaling in promoting viral lytic reactivation
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DOI：
10.1371/journal.ppat.1008156
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发表时间：
2019-12-01
期刊：
影响因子：
6.7
通讯作者：
Qin, Zhiqiang
中科院分区：
文献类型：
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作者：
Chen, Jungang;Dai, Lu;Qin, Zhiqiang
作为一种主要的致癌人类疱疹病毒，KSHV感染导致几种癌症，主要见于免疫功能低下的患者。目前，仍然缺乏有效的抗病毒治疗。“老药新技”的方法可能作为一个可行的策略，高通量筛选新的药物对裂解复制的KSHV。在这里，我们筛选了FDA批准的药物库，并确定了15种新的抗KSHV药物。有趣的是，这些候选人中有几个靶向组胺受体，这意味着组胺相关的信号转导参与了KSHV的裂解复制和再激活。这种参与是直接证明使用拮抗剂和激动剂的具体个别组胺受体以及RNAi。还鉴定了组胺介导的促进KSHV裂解复制所需的下游信号通路。来自HIV+患者队列的临床数据证实了HIV+/KSHV+患者微环境中组胺的相关性和升高。因此，我们的研究结果为开发抗KSHV治疗提供了新的线索，并确定了新的机制，通过组胺和相关的信号通路作为重要的宿主因子，促进KSHV裂解再激活和pathogenicity.Kaposi肉瘤相关疱疹病毒（KSHV）导致几种人类癌症，如Kaposi肉瘤（KS）和原发性渗出性淋巴瘤（PEL）。目前KSHV感染和病毒相关疾病的治疗选择有时是无效的，因此，迫切需要更有效的抗病毒药物。作为一种疱疹病毒，裂解性复制对于KSHV的发病机制和肿瘤发生至关重要。在这项研究中，我们已经建立了一个高通量的筛选试验，通过使用诱导KSHV+细胞系，iSLK.219。在筛选了由1280种食品和药物管理局（FDA）批准的药物组成的化合物库后，鉴定了15种有效抑制KSHV病毒粒子产生的命中化合物，其中大多数从未报道过具有抗KSHV活性。有趣的是，这些药物中有3种靶向组胺受体或信号传导。我们的数据进一步证实了靶向不同组胺受体（HxR）的拮抗剂对诱导的iSLK. 219或BCBL-1细胞的KSHV裂解性复制显示出优异的抑制作用。相比之下，组胺和HxR的特异性激动剂促进诱导的iSLK.219或KSHV感染的原代细胞的病毒裂解复制。机制研究表明，下游MAPK和PI 3 K/Akt信号通路是组胺/受体介导的促进KSHV裂解复制所必需的。iSLK.219细胞中HxR的直接敲低有效地阻断了诱导期间病毒裂解基因的表达。使用来自一组HIV+患者的样本，我们发现KSHV+组的血浆和唾液中的组胺水平比KSHV-组高得多。总之，我们的数据已经确定了新的抗KSHV药物，并提供了新的见解宿主因子的分子基础，有助于裂解复制和再激活这种致癌疱疹病毒。
Author summary As a major oncogenic human herpesviruses, KSHV infection causes several cancers mostly seen in immunocompromised patients. Currently, effective antiviral treatments are still lacking. The "old drugs, new tricks" approach may serve as a feasible strategy for high-throughput screening of novel agents against lytic replication of KSHV. Here we screened an FDA-approved drug library and identified 15 new anti-KSHV agents. Interestingly, several of these candidates target histamine receptors, implying the involvement of histamine-related signaling in lytic replication and reactivation of KSHV. This involvement was directly demonstrated using antagonists and agonists specific for individual histamine receptors as well as RNAi. The downstream signaling pathways required for histamine-mediated promotion of KSHV lytic replication was also identified. Clinical data from a cohort of HIV+ patients confirm the relevance and elevation of histamine in the microenvironment of HIV+/KSHV+ patients. Thus, our findings provide new clues for developing anti-KSHV treatments, and identify novel mechanisms through which histamine and related signaling pathways function as important host factors facilitating KSHV lytic reactivation and pathogenesis.Kaposi's sarcoma-associated herpesvirus (KSHV) causes several human cancers, such as Kaposi's sarcoma (KS) and primary effusion lymphoma (PEL). Current treatment options for KSHV infection and virus associated diseases are sometimes ineffective, therefore, more effectively antiviral agents are urgently needed. As a herpesvirus, lytic replication is critical for KSHV pathogenesis and oncogenesis. In this study, we have established a high-throughput screening assay by using an inducible KSHV+ cell-line, iSLK.219. After screening a compound library that consisted of 1280 Food and Drug Administration (FDA)-approved drugs, 15 hit compounds that effectively inhibited KSHV virion production were identified, most of which have never been reported with anti-KSHV activities. Interestingly, 3 of these drugs target histamine receptors or signaling. Our data further confirmed that antagonists targeting different histamine receptors (HxRs) displayed excellent inhibitory effects on KSHV lytic replication from induced iSLK.219 or BCBL-1 cells. In contrast, histamine and specific agonists of HxRs promoted viral lytic replication from induced iSLK.219 or KSHV-infected primary cells. Mechanistic studies indicated that downstream MAPK and PI3K/Akt signaling pathways were required for histamine/receptors mediated promotion of KSHV lytic replication. Direct knockdown of HxRs in iSLK.219 cells effectively blocked viral lytic gene expression during induction. Using samples from a cohort of HIV+ patients, we found that the KSHV+ group has much higher levels of histamine in their plasma and saliva than the KSHV- group. Taken together, our data have identified new anti-KSHV agents and provided novel insights into the molecular bases of host factors that contribute to lytic replication and reactivation of this oncogenic herpesvirus.