Profiling Peripheral Blood with an Optimized, Multiplexed, Single-cell Multiome Approach Supports an Insulin-driven...

Area of Science:

• Genomics
• Immunology
• Metabolic disease research

Background:

• Single-cell multiomic techniques like RNA sequencing (RNA-seq) and Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq) are vital for studying gene regulation.
• Current single-cell methods are complex and costly, hindering population-scale studies.
• Developing cost-effective, scalable single-cell investigations is crucial for population genomics.

Purpose of the Study:

• To adapt and optimize a multiomic protocol for sample multiplexing, creating multiplexed TEA-seq (mTEA-seq).
• To investigate the molecular signature of asthma associated with early-life elevated insulin levels.
• To identify sex-specific transcriptional and epigenetic alterations in adult blood cells related to childhood insulin levels and asthma.

Main Methods:

• Adaptation and optimization of the Transcriptome, Epitope, and ATAC sequencing (TEA-seq) protocol to create multiplexed TEA-seq (mTEA-seq).
• Analysis of blood samples from 54 adult subjects in the Tucson Children's Respiratory Study (TCRS) cohort.
• Application of a Bayesian approach for analyzing transcriptional and epigenetic data, including cell type-specific cis-QTL mapping.

Main Results:

• Male asthmatics with high childhood insulin showed significant immune transcriptional and epigenetic changes into adulthood.
• Male non-asthmatics with high childhood insulin exhibited epigenetic perturbations but not transcriptional changes.
• Epigenetic signals were cell-type-specific, with CD14+ monocytes showing common insulin-associated chromatin remodeling and NK cells displaying unique reprogramming patterns based on asthma status.

Conclusions:

• Childhood insulin-associated asthma has a distinct molecular signature characterized by persistent, metabolically-driven alterations in immune cells.
• The mTEA-seq protocol enables population-scale single-cell multiomic investigations.
• Findings suggest potential targets like HLA-DQB1 and AHI for understanding and treating insulin-associated asthma.