Kaposi's Sarcoma-Associated Herpesvirus Infection Induces the Expression of Neuroendocrine Genes in Endothelial Cells.
Kaposi's Sarcoma-Associated Herpesvirus Infection Induces the Expression of Neuroendocrine Genes in Endothelial Cells.
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卡波西肉瘤相关疱疹病毒感染诱导内皮细胞神经内分泌基因的表达。
DOI：
10.1128/jvi.01692-19
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发表时间：
2020
影响因子：
5.4
通讯作者：
Chandran,Bala
中科院分区：
文献类型：
--
作者：
ValiyaVeettil,Mohanan;Krishna,Gayathri;Roy,Arunava;Ghosh,Anandita;Dutta,Dipanjan;Kumar,Binod;Chakraborty,Sayan;Anju,ThoppilRaveendran;Sharma-Walia,Neelam;Chandran,Bala
卡波西肉瘤相关疱疹病毒（KSHV）与免疫功能低下个体的内皮卡波西肉瘤（KS）在病因学上相关。KS病变细胞与神经内分泌（NE）癌表现出许多相似之处，如高度血管化和红色/紫色肿瘤病变、梭形细胞、经典癌基因在肿瘤发展中的作用不显著、生物活性胺的释放和肿瘤的惰性生长。然而，KS病变内皮细胞与神经内分泌肿瘤相似的机制基础仍然未知。新一代测序和生物信息学分析表明，KSHV潜伏感染的内皮细胞表达几种神经元和NE基因，用活的和UV灭活的KSHV重新感染原代真皮内皮细胞表明，病毒基因表达负责上调五个选定的NE基因（肾上腺髓质素2 [ADM 2]、组胺受体H1 [HRH 1]、神经元特异性烯醇化酶[NSE] [ENO 2]、神经元蛋白基因产物9.5 [PGP 9.5]和生长抑素受体1 [SSTR 1]）。免疫荧光和免疫组织化学检测显示，NE基因HRH 1和NSE/ENO 2在KSHV感染的KS组织样本和KS内脏组织微阵列中的稳健表达。进一步分析表明，KSHV潜伏开放阅读框K12（ORFK 12）基因（kaposin A）介导的宿主REST/NRSF（RE 1-silencing transcription factor/neuron-restrictive silencer factor，RE 1-silencing factor/neuron-restrictive silencer factor）蛋白（一种神经元基因转录抑制蛋白）的降低，是导致NE基因在感染内皮细胞中表达的原因。在KSHV感染的细胞中观察到的NE基因表达在未感染的内皮细胞中通过ORFK 12的外源性表达和通过用REST抑制剂X5050处理细胞而重现。当CRISPR敲除神经活性配体激活受体HRH 1和抑制性SSTR 1时，HRH 1敲除（KO）显著抑制细胞增殖，而SSTR 1 KO诱导细胞增殖，因此表明HRH 1和SSTR 1可能在调节KSHV感染的内皮细胞增殖中相互抵消。这些结果表明，KS病变细胞的神经内分泌肿瘤的相似性可能是由于KSHV感染诱导的非神经元内皮细胞转化为具有神经内分泌功能的细胞。这些研究表明神经内分泌途径基因在KSHV感染的内皮细胞的病理生物学特征中的潜在作用，包括通过表达免疫特权神经元位点NE基因逃避宿主免疫系统的潜在机制，和NE基因可能作为KSHV感染的KS病变内皮细胞的标志物，以及控制KS病变的新的治疗靶点。相关疱疹病毒（KSHV）在感染的人类宿主中潜伏期间操纵几种细胞途径以获得其存活优势。在这里，我们证明，KSHV感染上调神经元和神经内分泌（NE）的功能，是NE肿瘤的特点，无论是在体外和KS患者组织和异质性的神经内分泌受体在KSHV感染的细胞增殖具有相反的作用基因的表达。KSHV对NE基因的诱导也可能提供潜在的生存优势，因为在免疫特权部位（如内皮细胞上的神经元）的蛋白质表达可能是逃避宿主免疫监视功能的途径。NE基因产物可作为KSHV的标记物。
Kaposi’s sarcoma-associated herpesvirus (KSHV) is etiologically associated with endothelial Kaposi’s sarcoma (KS) in immunocompromised individuals. KS lesion cells exhibit many similarities to neuroendocrine (NE) cancers, such as highly vascular and red/purple tumor lesions, spindle-shaped cells, an insignificant role for classic oncogenes in tumor development, the release of bioactive amines, and indolent growth of the tumors. However, the mechanistic basis for the similarity of KS lesion endothelial cells to neuroendocrine tumors remains unknown. Next-generation sequencing and bioinformatics analysis in the present study demonstrate that endothelial cells latently infected with KSHV express several neuronal and NE genes.De novoinfection of primary dermal endothelial cells with live and UV-inactivated KSHV demonstrated that viral gene expression is responsible for the upregulation of five selected NE genes (adrenomedullin 2 [ADM2], histamine receptor H1 [HRH1], neuron-specific enolase [NSE] [ENO2], neuronal protein gene product 9.5 [PGP9.5], and somatostatin receptor 1 [SSTR1]). Immunofluorescence and immunohistochemistry examinations demonstrated the robust expression of the NE genes HRH1 and NSE/ENO2 in KSHV-infected KS tissue samples and KS visceral tissue microarrays. Further analysis demonstrated that KSHV latent open reading frame K12 (ORFK12) gene (kaposin A)-mediated decreased host REST/NRSF (RE1-silencing transcription factor/neuron-restrictive silencer factor) protein, a neuronal gene transcription repressor protein, is responsible for NE gene expression in infected endothelial cells. The NE gene expression observed in KSHV-infected cells was recapitulated in uninfected endothelial cells by the exogenous expression of ORFK12 and by the treatment of cells with the REST inhibitor X5050. When the neuroactive ligand-activating receptor HRH1 and inhibitory SSTR1 were knocked out by CRISPR, HRH1 knockout (KO) significantly inhibited cell proliferation, while SSTR1 KO induced cell proliferation, thus suggesting that HRH1 and SSTR1 probably counteract each other in regulating KSHV-infected endothelial cell proliferation. These results demonstrate that the similarity of KS lesion cells to neuroendocrine tumors is probably a result of KSHV infection-induced transformation of nonneuronal endothelial cells into cells with neuroendocrine features. These studies suggest a potential role of neuroendocrine pathway genes in the pathobiological characteristics of KSHV-infected endothelial cells, including a potential mechanism of escape from the host immune system by the expression of immunologically privileged neuronal-site NE genes, and NE genes could potentially serve as markers for KSHV-infected KS lesion endothelial cells as well as novel therapeutic targets to control KS lesions.IMPORTANCEKaposi’s sarcoma-associated herpesvirus (KSHV) manipulates several cellular pathways for its survival advantage during its latency in the infected human host. Here, we demonstrate that KSHV infection upregulates the expression of genes related to neuronal and neuroendocrine (NE) functions that are characteristic of NE tumors, bothin vitroand in KS patient tissues and the heterogeneity of neuroendocrine receptors having opposing roles in KSHV-infected cell proliferation. Induction of NE genes by KSHV could also provide a potential survival advantage, as the expression of proteins at immunologically privileged sites such as neurons on endothelial cells may be an avenue to escape host immune surveillance functions. The NE gene products identified here could serve as markers for KSHV …