Dirvin Brooke, Noh Heeju, Tomassoni Lorenzo, Cao Danting, Zhou Yizhuo, Ke Xiangyi, Qian Jun, Jangra Sonia, Schotsaert Michael, García-Sastre Adolfo, Karan Charles, Califano Andrea, Cardoso Wellington V
Columbia Center for Human Development, Columbia University Irving Medical Center, New York, NY USA 10032.
Department of Genetics and Development, Columbia University Irving Medical Center, New York, NY, USA 10032.
bioRxiv. 2024 Dec 9:2024.10.11.617898. doi: 10.1101/2024.10.11.617898.
Although the impact of SARS-CoV-2 in the lung has been extensively studied, the molecular regulators and targets of the host-cell programs hijacked by the virus in distinct human airway epithelial cell populations remain poorly understood. This is in part ascribed to the use of nonprimary cell systems, overreliance on single-cell gene expression profiling that does not ultimately reflect protein activity, and bias toward the downstream effects rather than their mechanistic determinants. Here we address these issues by network-based analysis of single cell transcriptomic profiles of pathophysiologically relevant human adult basal, ciliated and secretory cells to identify master regulator (MR) protein modules controlling their SARS-CoV-2-mediated reprogramming. This uncovered chromatin remodeling, endosomal sorting, ubiquitin pathways, as well as proviral factors identified by CRISPR analyses as components of the host response collectively or selectively activated in these cells. Large-scale perturbation assays, using a clinically relevant drug library, identified 11 drugs able to invert the entire MR signature activated by SARS-CoV-2 in these cell types. Leveraging MR analysis and perturbational profiles of human primary cells represents a novel mechanism-based approach and resource that can be directly generalized to interrogate signatures of other airway conditions for drug prioritization.
尽管严重急性呼吸综合征冠状病毒2(SARS-CoV-2)对肺部的影响已得到广泛研究,但在不同的人类气道上皮细胞群体中,该病毒劫持的宿主细胞程序的分子调节因子和靶点仍知之甚少。部分原因在于使用了非原代细胞系统,过度依赖最终无法反映蛋白质活性的单细胞基因表达谱,以及偏向于下游效应而非其机制决定因素。在这里,我们通过对病理生理相关的人类成年基底细胞、纤毛细胞和分泌细胞的单细胞转录组谱进行基于网络的分析来解决这些问题,以识别控制其SARS-CoV-2介导的重编程的主调节因子(MR)蛋白模块。这揭示了染色质重塑、内体分选、泛素途径,以及通过CRISPR分析确定的作为宿主反应组成部分的前病毒因子,这些因子在这些细胞中共同或选择性地被激活。使用临床相关药物库进行的大规模扰动试验,确定了11种能够逆转SARS-CoV-2在这些细胞类型中激活的整个MR特征的药物。利用人类原代细胞的MR分析和扰动谱代表了一种基于机制的新方法和资源,可直接推广用于询问其他气道疾病的特征以进行药物优先级排序。