Assay of Genome-Wide Transcriptome and Secreted Proteins on the Same Single Immune Cells by Microfluidics and RNA...

Area of Science:

• Immunology
• Molecular Biology
• Bioinformatics

Background:

• Immune cell heterogeneity poses challenges in studying gene expression related to specific functions like cytokine production.
• Existing methods struggle to simultaneously analyze protein and gene expression at the single-cell level.

Purpose of the Study:

• To develop a technology enabling simultaneous analysis of protein and genome-wide transcriptome from the same single immune cell.
• To identify gene expression patterns associated with specific cellular functions, such as cytokine secretion.

Main Methods:

• Development of a splittable single-cell microchip integrating antibody array for protein detection.
• Subsequent whole-transcriptome sequencing of the same single cells after protein profiling.
• Application of bioinformatics algorithms for data analysis.

Main Results:

• Discovery of a subgroup of coexpressed genes correlated with Tumor Necrosis Factor alpha (TNFα) secretion in mouse macrophages.
• Demonstration of a technology capable of linking protein secretion profiles with cellular transcriptomes.
• Identification of novel gene expression networks regulating immune cell function.

Conclusions:

• The developed microchip technology overcomes limitations in studying immune cell heterogeneity and function.
• This approach provides unprecedented insights into immune system regulation mechanisms.
• The technology holds potential for advancing disease treatment and drug discovery by understanding immune responses.