Dynamic Transcriptional Programs During Single NK Cell Killing: Connecting Form to Function in Cellular Immunotherapy

Area of Science:

• Immunology
• Cancer Biology
• Molecular Biology

Background:

• Natural killer (NK) cell therapies show promise for cancer treatment, but progress for solid tumors is slow.
• Understanding the intracellular signaling pathways that drive NK cell killing is crucial for developing effective therapies.

Purpose of the Study:

• To correlate transcriptomic data with functional output in NK cell killing using a combined single-cell RNA sequencing and live cell imaging approach.
• To identify intracellular signaling molecules that drive the NK cell killing phenotype.

Main Methods:

• Utilized a model system of NK-92 cells killing HeLa cervical cancer cells.
• Combined single-cell RNA sequencing (10× Genomics) with live cell imaging and reporter constructs.
• Inferred transcription factor activity using Viper and DoRothEA R packages.

Main Results:

• NK cells exhibited heterogeneous killing activity over 10 hours.
• Nuclear Factor Kappa B (NF-κB) activity was identified as a potential driver of NK cell function.
• NF-κB activity significantly correlated with past NK cell killing activity, while STAT1 and MYC did not.

Conclusions:

• The integration of live cell imaging of transcription factor activity with single-cell RNA sequencing effectively links transcriptomic data to functional phenotypes.
• This approach enhances the understanding of NK cell-mediated cytotoxicity, paving the way for improved cancer immunotherapies.