microRNA‐34‐5p encoded by Spodoptera frugiperda regulates the replication and infection of Autographa californica multiple nucleopolyhedrovirus by targeting odv‐e66, ac78 and ie2

Article

作者: Fu, Yuejun ; Jia, Qiaojin

Abstract:

BACKGROUND:

Spodoptera frugiperda is one of the significant migratory pests in the Global Alert issued by the Food and Agriculture Organization of the United Nations. As an insect‐specific microbial insecticide, baculovirus has been used to control various pests. MicroRNA‐34‐5p (miR‐34‐5p) is involved in regulating growth, reproduction and innate immunity to pathogens in insects, playing an essential role in host–virus interactions. In this study, we explored the critical function of miR‐34‐5p encoded by S. frugiperda in the anti‐Autographa californica multiple nucleopolyhedrovirus (AcMNPV), providing a reference for the design of a miR‐34‐5p target biopesticide against S. frugiperda and a theoretical basis for the wide application of microRNAs (miRNAs) in green pest control technology.

RESULTS:

We focused on miR‐34‐5p identified as downregulated in Sf9 cells and S. frugiperda larvae infected by AcMNPV. The regulatory function of miR‐34‐5p in AcMNPV–S. frugiperda interactions was studied by transfecting synthetic mimics and inhibitors, and constructing recombinant bacmids with miR‐34‐5p overexpression. miR‐34‐5p inhibited the production of infectious budded virions at the cellular and insect levels, inhibited the replication of the viral DNA and glucose metabolism, and increased the transcription of the antimicrobial peptide gloverin. Furthermore, the virus genes odv‐e66, ac78 and ie2 were shown to be direct targets.

CONCLUSION:

We systematically revealed the mechanism by which miR‐34‐5p is involved in the insect antiviral process. miR‐34‐5p inhibited the replication and infection of AcMNPV by directly targeting AcMNPV genes, especially ac78 and ie2. Our study provides a new direction and thinking for the prevention and green control of lepidopteran pests. © 2022 Society of Chemical Industry.

2014-10-01·Virus research3区 · 医学

Disruption of the baculovirus core gene ac78 results in decreased production of multiple nucleocapsid-enveloped occlusion-derived virions and the failure of primary infection in vivo

3区 · 医学

Article

作者: Yang, Kai ; Yuan, Mei-Jin ; Li, Sai-Nan ; Wang, Jin-Yu

The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ac78 gene is one of the baculovirus core genes. Recent studies showed that ac78 is essential for budded virion (BV) production and the embedding of occlusion-derived virion (ODV) into occlusion body during the AcMNPV life cycle. Here, we report that an ac78-knockout AcMNPV (vAc78KO) constructed in this study had different phenotypes than those described in the previous studies. A few infectious BVs were detected using titer assays, immunoblot analyses and plaque assays, indicating that ac78 is not essential for BV formation. Electron microscopy confirmed that the ac78 deletion did not affect nucleocapsid assembly and ODV formation. However, the numbers of multiple nucleocapsid-enveloped ODVs and ODV-embedded occlusion bodies were significantly decreased. Subsequently, the highly conserved amino acid residues 2-25 and 64-88 of Ac78, which are homologous to an oxidoreductase and cytochrome c oxidase, respectively, were demonstrated to play a crucial role in the morphogenesis of multiple nucleocapsid-enveloped ODV. Immunoblot analysis found that Ac78 was an ODV envelope-associated protein. Consistently, amino acid residues 56-93 of Ac78 were identified as an inner nuclear membrane sorting motif, which may direct the localization of Ac78 to the ODV envelope. In vivo infectivity assays showed that the occlusion bodies of vAc78KO were unable to establish primary infection in the midgut of Trichoplusia ni larvae. Taken together, our results suggest that ac78 plays an important role in BV production and proper multiple nucleocapsid-enveloped ODV formation, as well as AcMNPV primary infection in vivo.

2013-08-01·Journal of virology2区 · 医学

The Autographa californica Multiple Nucleopolyhedrovirus ORF78 Is Essential for Budded Virus Production and General Occlusion Body Formation

2区 · 医学

Article

作者: Lee, Seok Hee ; Lee, Joo Hyun ; An, Saes Byeol ; Kim, Woo Jin ; Jin, Byung Rae ; Woo, Soo Dong ; Tao, Xue Ying ; Liu, Qin ; Kim, Song Eun ; Choi, Jae Young ; Je, Yeon Ho

ABSTRACT:

ORF78 ( ac78 ) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a baculovirus core gene of unknown function. To determine the role of ac78 in the baculovirus life cycle, an AcMNPV mutant with ac78 deleted, Ac78KO, was constructed. Quantitative PCR analysis revealed that ac78 is a late gene in the viral life cycle. After transfection into Spodoptera frugiperda cells, Ac78KO produced a single-cell infection phenotype, indicating that no infectious budded viruses (BVs) were produced. The defect in BV production was also confirmed by both viral titration and Western blotting. However, viral DNA replication was unaffected, and occlusion bodies were formed. An analysis of BVs and occlusion-derived viruses (ODVs) revealed that AC78 is associated with both forms of the virions and is an envelope structural protein. Electron microscopy revealed that AC78 also plays an important role in the embedding of ODV into the occlusion body. The results of this study demonstrate that AC78 is a late virion-associated protein and is essential for the viral life cycle.

项与 AC0078 相关的新闻（医药）

2021-01-08

·药融圈

【产业观察】以为小适应症，结果TRK新药大热门

精准医疗的典范 NTRK，即神经营养因子受体酪氨酸激酶（Neuro Trophin Receptor Kinase），由高度同源性的激酶TRK-A、TRK-B、TRK-C组成，属于NTRK基因编码。NTRK基因融合可以发生在身体的任何部位，因此TRK融合癌症可能出现在多种成人和儿童实体瘤中，包括乳腺类似物分泌癌（MASC）、结肠癌、肺癌，胰腺癌、甲状腺癌、以及各种肉瘤，这种基因融合可能影响超过60％的患有某些罕见癌症的成人和儿童患者。在非小细胞肺癌和结直肠癌中有1-3%的患者发生NTRK基因融合；在罕见的癌症中，如婴儿纤维肉瘤，乳腺类似物分泌型癌症等，高达90%以上的患者发生了NTRK基因融合。ALK及ROS1主要在发生在非小细胞肺癌（发生率分别约为7%及1%）。本质上，本靶点领域为相对精准的治疗领域。目前，美国FDA已经批准两款不分组织，不分瘤种的小分子TRK抑制剂，分别是Loxo oncology/拜耳于2018年11月28日获批的拉罗替尼（Larotrectinib，商品名“Vitrakvi”）和罗氏/Ignyta于2019年8月16日获批的恩曲替尼（Entrectinib，商品名“Rozlytrek”）。此外海外有多款二代新药在研，不一一赘述。国内TRK抑制剂在研火热可能一些企业申报了临床试验才知道，原来这个“床”这么拥挤；有的企业立项时，本以为小适应症，结果TRK抑制剂（多靶点）成为新药大热门。也罢，共同为中国乃至全球患者提供更优质的新药，毕竟这么多申报还是可以有些积极临床试验结果的。不过最担忧的可能是部分细分临床试验的病人够不够用。一.TRK抑制剂（多靶点）部分申报二.除以上企业外其他申报TRK抑制剂（多靶点）的有再鼎医药，Repotrectinib胶囊,一种抑制ROS1、TRK和ALK活性的广谱新一代TKI；先声药业，SIM1803-1A片，新一代选择性针对TRK A/B/C和ROS1的激酶抑制剂；绿叶制药（南京康海磷脂生物），LPM4870108片（LY01018）；江苏威凯尔，VC004胶囊；复星复创医药，FCN-011胶囊；药融圈统计，目前合计约13家都在或者将在国内展开临床试验。三.其他TRK抑制剂（多靶点）在研艾森生物，AC0078；成都海创药业，NTRK专利；中科院合肥研究院健康所，IHMT-TRK-284；南京柯菲平，H103，已经提交国内发明专利；等等。四.部分案例 1.葆元医药，AB-106（Taletrectinib），ROS1/NTRK抑制剂，在中国，正在开展一项Ⅱ期、多中心、单臂、开放的临床研究（上市为目的）用于一线和二线治疗携带ROS1融合基因的局部进展或全身转移的晚期非小细胞肺癌（NSCLC）；和一项Ⅱ期、多中心、开放、篮式研究用于治疗携带NTRK 融合基因的局部进展或全身转移的实体瘤；，可能将抢得临床资源和市场先机。 AB-106具有以下特点：（1）对ROS1及NTRK1-3具有高效抑制作用，对克唑替尼的主要耐药位点ROS1 G2032R同样具有高效抑制作用；（2）安全性好，应答率高，有效持续时间长，带给病人更长的无疾病进展生存期；（3）对克唑替尼治疗失败的ROS1阳性非小细胞肺癌患者及脑转移患者也有疗效。此前在2018年12月18日，日本第一三共株式会社和美国葆元生物医药科技公司（下称美国葆元）共同宣布，双方就一款用于癌症治疗的ROS1/NTRK抑制剂（DS-6051）签署了全球独家许可协议。根据协议，第一三共将DS-6051的全球开发，生产和商业化权利独家许可给了美国葆元。第一三共将获得一笔首付款，以及未来将获得一系列临床，监管审批和销售里程碑付款，以及一定比例的全球净销售利润提成。具体财务条款未被披露。 2.于国内企业而言，进展较快的有：成都先导的HG030片为国内首家获准临床研究的第二代TRK小分子抑制剂，临床拟用于治疗NTRK或ROS1基因融合的实体瘤患者。此前在2020年11月10日，成都先导宣布与广州白云山医药集团股份有限公司签署转让合作协议，成都先导将新药项目代号为HG030的产品在中国大陆地区的全部权利转让给白云山，后续研发与商业化将有白云山独立推进并实施。根据协议约定，白云山将获得HG030在中国大陆后续开发及商业化的排他性权益，并且支付成都先导3,800万元人民币的转让费。随着项目的推进，成都先导还可能收到一系列的里程碑付款，以及当项目在中国大陆推进到临床阶段和产品上市后的销售提存。成都先导仍保留HG030产品中国大陆以外所有区域的全部权益。 3.2020年7月，再鼎医药宣布与Turning Point Therapeutics达成合作，获得Repotrectinib在大中华区的独家开发及商业化权。根据协议条款，Turning Point Therapeutics将获得2500万美元的现金首付款，最高至1.51亿美元的潜在开发、注册和基于销售的里程碑付款，以及特许权使用费。 2020年11月，Turning Point Therapeutics和再鼎医药联合申报的Repotrectinib胶囊获得一项临床默示许可，拟开发治疗携带ALK、ROS1或NTRK1-3重排的晚期实体瘤。更多药物信息，请查询药融云数据 www.pharnexcloud.com 参考： NMPA/CDE；药融云数据； FDA/EMA/PMDA；相关公司公开披露； Nature Reviews | Clinical Oncology.https://doi.org/10.1038/s41571-018-0113-0； Miriam Grazia Ferrara, Cancers (Basel). 2020 May; 12(5): 1196； https://anhearttherapeutics.com/2018/12/18/20191218-2/； https://www.hitgen.com/； http://www.zailaboratory.com/ch/about/index.aspx； https://cn.innocarepharma.com/media/1451/%E8%AF%BA%E8%AF%9A%E5%81%A5%E5%8D%8E2020%E5%B9%B4%E4%B8%AD%E6%9C%9F%E4%B8%9A%E7%BB%A9%E9%99%88%E8%BF%B0.pdf；等等。【关于药融圈】药融圈围绕我国生物医药产业链，针对生物医药大数据、技术和资本投资、药融园(产业园)等开展系列系统性工作，促进我国生物医药产业健康发展，完善产业链，共同面对全球合作和竞争。点分享点点赞点在看