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+信号传导是细胞功能的广泛保守调节剂，因此许多病毒已将机制发展为 利用CA2+信号通路以促进病毒复制。轮状病毒（RV），这仍然是 全球儿童的急性胃肠炎和腹泻死亡是一个原型的例子。 RV感染 诱导胞质Ca2+的全球增加，这既破坏宿主细胞生理，又有促进RV 复制。虽然全球CA2+失调长期以来一直是RV感染的标志，但信号的不同类型 维持这种失调，从而促进复制和发病机理，尚未得到充分识别。 最近，我们的实验室解决了一种新型RV诱导的Ca2+信号，称为细胞间CA2+波（ICW），这是 与病毒剥削和宿主恢复原状息息相关。 ICW是由旁分泌ADP从释放中介导的 感染的细胞与邻近细胞上的P2Y1受体结合，引起胞质Ca2+通量。通过这个 机理，RV不仅在受感染的细胞中，而且在相邻的未感染细胞中也使CA2+失调。我们发现 该P2Y1介导的信号对于影响疾病的宿主反应至关重要，包括5-羟色胺，氯化物， 和流体分泌，以及免疫介质的表达。在初步研究中，我们还发现 ICW增强了MAPK激活和干细胞标记，这有助于恢复原状。但是，抑制ICW 减少了RV的脱落，这表明ICW也有助于RV复制。重要的是，我们的初步数据 证明RV非结构蛋白4（NSP4）的重组表达，内质网局部 病毒Ca2+通道足以诱导ICW。鉴于此，我们的核心假设是CA2+异常 由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 宿主对RV感染的上皮反应；但是，RV利用此P2Y1发出的信号传导到周围的原始 细胞以进行更大的复制。为了检验这一假设，我们将（AIM 1）表征RV诱导的ADP释放 受感染细胞的途径（AIM 2）表征了P2Y1介导的增殖和恢复原状反应 肠道中的RV感染，（AIM 3）确定RV Co-Opts P2Y1信号如何增加病毒复制和 传播。鉴于广泛的肠道病原体破坏了Ca2+信号，这些目标的完成将会 对宿主/病原体武器竞赛产生新的机械洞察力，对病原体识别的新理解 途径，并确定潜在地作用抗病毒疗法的新细胞靶标。

项目成果

Gastrointestinal organoids in the study of viral infections.

病毒感染研究中的胃肠道类器官。

• DOI: 10.1152/ajpgi.00152.2022
• 发表时间: 2023
• 期刊: American journal of physiology. Gastrointestinal and liver physiology
• 作者: Gebert,JThomas;Scribano,Francesca;Engevik,KristenA;Perry,JacobL;Hyser,JosephM
• 通讯作者: Hyser,JosephM

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
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Engevik,Kristen;Kawagishi,Takahiro;Greenberg,Harry;Hyser,Joseph
• 通讯作者: Hyser,Joseph

Live Calcium Imaging of Virus-Infected Human Intestinal Organoid Monolayers Using Genetically Encoded Calcium Indicators.

使用基因编码钙指示剂对病毒感染的人肠类器官单层进行活钙成像。

• DOI: 10.3791/66132
• 发表时间: 2024
• 期刊: Journal of visualized experiments : JoVE
• 作者: Gebert,JThomas;Scribano,FrancescaJ;Engevik,KristenA;Hyser,JosephM
• 通讯作者: Hyser,JosephM

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。

• DOI: 10.1038/s41598-020-61112-0
• 发表时间: 2020
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Tomaro-Duchesneau,Catherine;LeValley,StephanieL;Roeth,Daniel;Sun,Liang;Horrigan,FrankT;Kalkum,Markus;Hyser,JosephM;Britton,RobertA
• 通讯作者: Britton,RobertA