The population context is a driver of the heterogeneous response of epithelial cells to interferons

Isogenic cells respond in a heterogeneous manner to interferon. Using a micropatterning approach combined with high-content imaging and spatial analyses, we characterized how the population context (position of a cell with respect to neighboring cells) of epithelial cells affects their response to interferons. We identified that cells at the edge of cellular colonies are more responsive than cells embedded within colonies. We determined that this spatial heterogeneity in interferon response resulted from the polarized basolateral interferon receptor distribution, making cells located in the center of cellular colonies less responsive to ectopic interferon stimulation. This was conserved across cell lines and primary cells originating from epithelial tissues. Importantly, cells embedded within cellular colonies were not protected from viral infection by apical interferon treatment, demonstrating that the population context-driven heterogeneous response to interferon influences the outcome of viral infection. Our data highlights that the behavior of isolated cells does not directly translate to their behavior in a population, placing the population context as one important factor influencing heterogeneity during interferon response in epithelial cells.

同基因细胞（isogenic cells）对干扰素（interferon）的应答呈现异质性特征。本研究结合微图案化（micropatterning）技术、高内涵成像（high-content imaging）与空间分析（spatial analyses），系统阐明了上皮细胞（epithelial cells）的群体背景——即细胞相对于相邻细胞的位置——如何影响其对干扰素的应答。我们发现，细胞集落（cellular colonies）边缘的细胞相较于集落内部的嵌入细胞，其应答活性更强。进一步研究表明，这种干扰素应答的空间异质性源于干扰素受体的极性基底外侧分布，使得细胞集落中心的细胞对外源性干扰素刺激的应答能力更弱。该现象在上皮组织来源的细胞系与原代细胞中均保守存在。值得注意的是，通过顶侧施加干扰素无法保护细胞集落内部的细胞免受病毒感染，这表明群体背景介导的干扰素应答异质性会直接影响病毒感染的结局。本研究数据揭示：分离状态下的细胞行为无法直接推演至其在群体环境中的行为，从而明确群体背景是影响上皮细胞干扰素应答过程中异质性的关键因素之一。