Enteric virus exploitation of calcium signaling

肠道病毒利用钙信号传导

• 批准号: 10735222
• 负责人: Joseph M. Hyser
• 金额: $ 64.41万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-13 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10735222
• 关键词: 5 year old; Acute; Affect; Anions; Antidiarrheals; Antiviral Response; Antiviral Therapy; Autophagocytosis; Binding; Biochemical; Biological Response Modifiers; Calcium; Calcium Oscillations; Calcium Signaling; Cell Line; Cell physiology; Cells; Cessation of life; Child; Chlorides; Complex; Data; Diarrhea; Disease; Distal; Endoplasmic Reticulum; Enteral; Epithelium; Fluids and Secretions; Functional disorder; Funding; Gastroenteritis; Gastrointestinal tract structure; Genetic; Genetic Transcription; Goals; Human; Image; Imaging Techniques; Immune response; Infection; Intestines; Knock-out; Life; Link; MAP Kinase Gene; Measures; Mediating; Modeling; Molecular Biology; Nature; Nonstructural Protein; Organoids; Outcome; Pathogenesis; Pathway interactions; Physiological; Play; Proviruses; Purinoceptor; Recombinants; Reticulum; Role; Rotavirus; Rotavirus Infections; Serotonin; Signal Pathway; Signal Transduction; Testing; Villus; Viral; Virus; Virus Replication; arms race; cell type; cellular targeting; clinically relevant; cytokine; enteric pathogen; enteric virus infection; gastrointestinal; imaging approach; insight; mortality; new therapeutic target; novel; paracrine; pathogen; programs; purinoceptor P2Y1; repaired; response; stem cell biomarkers; transcriptomics; vaccine access; virus host interaction

Ca2+ signaling is a broadly conserved regulator of cell function, thus many viruses have evolved mechanisms to exploit Ca2+ signaling pathways to promote viral replication. Rotavirus (RV), which remains the leading cause of acute gastroenteritis and diarrheal deaths in children worldwide, is an archetypal example of this. RV infection induces global increases in cytosolic Ca2+ which both disrupts host cell physiology and facilitates RV replication. While global Ca2+ dysregulation has long been a hallmark of RV infection, the distinct types of signals that sustain this dysregulation, and thereby facilitate replication and pathogenesis, have not been fully identified. Recently, our lab resolved a novel RV-induced Ca2+ signal known as an intercellular Ca2+ wave (ICW), which is critically linked to both viral exploitation and host restitution. ICWs are mediated by paracrine ADP release from infected cells which binds to the P2Y1 receptor on neighboring cells, eliciting a cytosolic Ca2+ flux. Through this mechanism, RV dysregulates Ca2+ not only in infected cells, but also in neighboring, uninfected cells. We found that this P2Y1-mediated signal is critical to host responses that influence disease, including serotonin, chloride, and fluid secretion, and the expression of immune mediators. In preliminary studies, we have also found that ICWs enhance MAPK activation and stem cell markers, which contribute to restitution. However, inhibiting ICWs reduces RV shedding, suggesting that ICWs also facilitate RV replication. Importantly, our preliminary data also show that recombinant expression of RV non-structural protein 4 (NSP4), an endoplasmic reticulum localized viral Ca2+ channel, is sufficient to induce ICWs. Given this, our Central Hypothesis is that aberrant Ca2+ signaling caused by RV NSP4 initiates ADP release triggering paracrine activation of P2Y1 to regulate host epithelial responses to RV infection; however, RV exploits this P2Y1 signaling to prime surrounding cells for greater replication. To test this hypothesis, we will (Aim 1) characterize the RV-induced ADP release pathway from infected cells, (Aim 2) characterize the P2Y1-mediated proliferative and restitution responses to RV infection in the gut, and (Aim 3) determine how RV co-opts P2Y1 signaling to increase virus replication and spread. Given that a broad range of enteric pathogens disrupt Ca2+ signaling, completion of these aims will generate new mechanistic insights into the host/pathogen arms race, new understanding of pathogen recognition pathways, and identify new cellular targets for potentially broadly acting antiviral therapies.

Ca2+ 信号传导是一种广泛保守的细胞功能调节因子，因此许多病毒已经进化出机制来 利用 Ca2+ 信号通路促进病毒复制。轮状病毒 (RV) 仍然是导致疾病的主要原因 全世界儿童的急性胃肠炎和腹泻死亡就是一个典型例子。右心室感染 诱导胞质 Ca2+ 整体增加，这既破坏宿主细胞生理学又促进 RV 复制。虽然全球 Ca2+ 失调长期以来一直是 RV 感染的标志，但不同类型的信号 维持这种失调，从而促进复制和发病机制的机制尚未完全确定。 最近，我们的实验室解决了一种新的 RV 诱导的 Ca2+ 信号，称为细胞间 Ca2+ 波 (ICW)，它是 与病毒利用和宿主恢复密切相关。 ICWs 由旁分泌 ADP 释放介导 受感染的细胞与邻近细胞上的 P2Y1 受体结合，引发胞质 Ca2+ 通量。通过这个 根据机制，RV 不仅会在感染细胞中失调 Ca2+，而且还会在邻近的未感染细胞中失调。我们发现 这种 P2Y1 介导的信号对于影响疾病的宿主反应至关重要，包括血清素、氯、 和液体分泌以及免疫介质的表达。在初步研究中，我们还发现 ICW 增强 MAPK 激活和干细胞标记，有助于恢复。然而，抑制 ICW 减少 RV 脱落，表明 ICW 也促进 RV 复制。重要的是，我们的初步数据还 显示 RV 非结构蛋白 4 (NSP4) 的重组表达，RV 非结构蛋白 4 是一种定位于内质网的蛋白。 病毒 Ca2+ 通道足以诱导 ICW。鉴于此，我们的中心假设是异常的 Ca2+ RV NSP4 引起的信号传导启动 ADP 释放，触发 P2Y1 的旁分泌激活以调节 宿主上皮对 RV 感染的反应；然而，RV 利用这种 P2Y1 信号向周围环境发出信号 细胞以实现更大程度的复制。为了检验这一假设，我们将（目标 1）描述 RV 诱导的 ADP 释放 来自受感染细胞的途径，（目标 2）表征 P2Y1 介导的增殖和恢复反应 肠道中的 RV 感染，以及（目标 3）确定 RV 如何利用 P2Y1 信号传导来增加病毒复制和 传播。鉴于多种肠道病原体会破坏 Ca2+ 信号传导，完成这些目标将 对宿主/病原体军备竞赛产生新的机制见解，对病原体识别有新的认识 途径，并确定潜在广泛作用的抗病毒疗法的新细胞靶标。

项目成果

Generation of Recombinant Rotavirus Expressing NSP3-UnaG Fusion Protein by a Simplified Reverse Genetics System.

通过简化的反向遗传学系统产生表达 NSP3-UnaG 融合蛋白的重组轮状病毒。

• 发表时间: 2019-12-15
• 作者: Philip, Asha A;Perry, Jacob L;Eaton, Heather E;Shmulevitz, Maya;Hyser, Joseph M;Patton, John T
• 通讯作者: Patton, John T

Rotavirus infection elicits host responses and amplifies viral replication via P2Y1 purinergic signaling.

轮状病毒感染通过 P2Y1 嘌呤能信号引发宿主反应并放大病毒复制。

• 发表时间: 2022-05
• 作者: Engevik, Kristen;Kawagishi, Takahiro;Greenberg, Harry;Hyser, Joseph
• 通讯作者: Hyser, Joseph

Gastrointestinal organoids in the study of viral infections.

病毒感染研究中的胃肠道类器官。

• 发表时间: 2023-01-01
• 作者: Gebert, J Thomas;Scribano, Francesca;Engevik, Kristen A;Perry, Jacob L;Hyser, Joseph M
• 通讯作者: Hyser, Joseph M

Discovery of a bacterial peptide as a modulator of GLP-1 and metabolic disease.

发现一种细菌肽作为 GLP-1 和代谢疾病的调节剂。

• 发表时间: 2020
• 影响因子: 4.6
• 作者: Tomaro;LeValley, Stephanie L;Roeth, Daniel;Sun, Liang;Horrigan, Frank T;Kalkum, Markus;Hyser, Joseph M;Britton, Robert A
• 通讯作者: Britton, Robert A

Host IP3R channels are dispensable for rotavirus Ca2+ signaling but critical for intercellular Ca2+ waves that prime uninfected cells for rapid virus spread.

宿主 IP3R 通道对于轮状病毒 Ca2 信号传导来说是可有可无的，但对于细胞间 Ca2 波至关重要，细胞间 Ca2 波为未感染细胞提供病毒快速传播的准备。

• 发表时间: 2024-01-16
• 影响因子: 6.4
• 作者: Perry, Jacob L;Scribano, Francesca J;Gebert, John T;Engevik, Kristen A;Ellis, Jenna M;Hyser, Joseph M
• 通讯作者: Hyser, Joseph M