Establishment of a human organoid-based evaluation system for assessing interspecies infection risk of animal-borne coronaviruses

The COVID-19 pandemic presents a major threat to global public health. Several lines of evidence have shown that the Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), along with two other highly pathogenic coronaviruses, SARS-CoV and Middle East Respiratory Syndrome (MERS-CoV) originated from bats. To prevent and control future coronavirus outbreaks, it is necessary to investigate the interspecies infection and pathogenicity risks of animal-related coronaviruses. Currently used infection models, including in vitro cell lines and in vivo animal models, fail to fully mimic the primary infection in human tissues. Here, we employed organoid technology as a promising new model for studying emerging pathogens and their pathogenic mechanisms. We investigated the key host-virus interaction patterns of five human coronaviruses (SARS-CoV-2 original strain, Omicron BA.1, MERS-CoV, HCoV-229E, and HCoV-OC43) in different human respiratory organoids. Five indicators, including cell tropism, invasion preference, replication activity, host response and virus-induced cell death, were developed to establish a comprehensive evaluation system to predict coronavirus interspecies infection and pathogenicity risks. Using this system, we further examined the pathogenicity and interspecies infection risks of three SARS-related coronaviruses (SARSr-CoV), including WIV1 and rRsSHC014S from bats, and MpCoV-GX from pangolins. Moreover, we found that cannabidiol, a non-psychoactive plant extract, exhibits significant inhibitory effects on various coronaviruses in human lung organoid. Cannabidiol significantly enhanced interferon-stimulated gene expression but reduced levels of inflammatory cytokines. In summary, our study established a reliable comprehensive evaluation system to analyse infection and pathogenicity patterns of zoonotic coronaviruses, which could aid in prevention and control of potentially emerging coronavirus diseases.

新型冠状病毒肺炎（COVID-19）大流行对全球公共卫生构成重大威胁。多项研究证据表明，严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）与另外两种高致病性冠状病毒——严重急性呼吸综合征冠状病毒（SARS-CoV）、中东呼吸综合征冠状病毒（MERS-CoV）——均起源于蝙蝠。为防控未来冠状病毒暴发，亟需开展动物源性冠状病毒跨物种感染及致病风险相关研究。当前主流的感染模型，包括体外细胞系与体内动物模型，均无法完全模拟人体组织的原发感染过程。本研究采用类器官（organoid）技术，将其作为研究新兴病原体及其致病机制的新型理想模型。我们针对五种人类冠状病毒——严重急性呼吸综合征冠状病毒2型原始毒株、奥密克戎BA.1亚型、中东呼吸综合征冠状病毒（MERS-CoV）、人冠状病毒229E（HCoV-229E）、人冠状病毒OC43（HCoV-OC43）——在不同人体呼吸道类器官中的关键宿主-病毒互作模式展开研究。本研究构建了涵盖细胞嗜性、入侵偏好、复制活性、宿主应答及病毒诱导细胞死亡五大指标的综合评估体系，用于预测冠状病毒跨物种感染与致病风险。依托该评估体系，我们进一步探究了三种严重急性呼吸综合征相关冠状病毒（SARSr-CoV）的致病性与跨物种感染风险，分别为源自蝙蝠的WIV1、rRsSHC014S，以及来自穿山甲的MpCoV-GX。此外，本研究发现非精神活性植物提取物大麻二酚（cannabidiol）对人体肺类器官中的多种冠状病毒具有显著抑制效果。大麻二酚可显著上调干扰素刺激基因（interferon-stimulated gene）的表达水平，同时降低炎症细胞因子（inflammatory cytokines）的分泌量。综上，本研究建立了一套可靠的综合评估体系，用于分析人畜共患冠状病毒的感染与致病模式，可为潜在新发冠状病毒病的防控工作提供有力支撑。