TNFR1 mediates heterogeneity in single-cell NF-κB activation

Chieh-Teng Cheng; Jye-Chian Hsiao; Alexander Hoffmann; Hsiung-Lin Tu

doi:10.1016/j.isci.2024.109486

TNFR1 mediates heterogeneity in single-cell NF-κB activation

iScience. 2024 Mar 11;27(4):109486. doi: 10.1016/j.isci.2024.109486. eCollection 2024 Apr 19.

Authors

Chieh-Teng Cheng^{1

2}, Jye-Chian Hsiao¹, Alexander Hoffmann³, Hsiung-Lin Tu^{1

2}

Affiliations

¹ Institute of Chemistry, Academia Sinica, Taipei 11529, Taiwan.
² Genome and Systems Biology Degree Program, Academia Sinica and National Taiwan University, Taipei 10617, Taiwan.
³ Department of Microbiology, Immunology and Molecular Genetics, and Institute for Quantitative and Computational Biosciences, University of California, Los Angeles, Los Angeles, CA 99093, USA.

Abstract

Nuclear factor kappa B (NF-κB) is a key regulator in immune signaling and is known to exhibit a digital activation pattern. Yet the molecular basis underlying the heterogeneity in NF-κB activation at single-cell level is not entirely understood. Here, we show that NF-κB activation in single cells is largely regulated by intrinsic differences at the receptor level. Using the genome editing and time-lapse imaging, we directly characterize endogenous TNFR1 dynamics and NF-κB activation from the same single cells. Total internal reflection fluorescence (TIRF) microscopy shows that endogenous TNFR1 forms pre-ligand clusters in the resting cells. Upon tumor necrosis factor (TNF) stimulation, the diffusion coefficient of membrane TNFR1 was significantly decreased and a substantial level of TNFR1 undergoes oligomerization to form trimers and hexamers. Moreover, multi-color cell imaging reveals that both digital and graded information processing regulate NF-κB activation across different TNFR1 expression levels. Our results indicate that single-cell NF-κB activation potential strongly correlates with its TNFR1 characteristics.

Keywords: Biological sciences; Cell biology; Molecular biology.