# α-Diversity analysis of hepatic transcriptome reveals distinct pathways in alcohol-associated hepatitis

## TL;DR

This study uses α-diversity analysis to uncover new pathways in alcohol-related liver disease that traditional methods miss.

## Contribution

The study introduces α-diversity as a novel analytical approach to reveal hidden transcriptomic changes in alcohol-associated hepatitis.

## Key findings

- ALD significantly reduces hepatic transcriptome α-diversity, linked to increased contribution of select genes.
- DSD identifies additional pathways like fatty acid oxidation and cholesterol metabolism not captured by DEG analysis.
- DSD more effectively distinguishes between alcoholic steatohepatitis and severe alcohol-associated hepatitis.

## Abstract

Next-generation sequencing can identify previously uncharacterized gene expression patterns in disease. Beyond differentially expressed gene (DEG) analysis, we investigated the ability of within-population diversity (α-diversity) of the transcriptome to reveal additional biological information in alcohol-associated liver disease (ALD), comparing differential Shannon diversity (DSD) to transcriptome heterogeneity changes. RNA sequencing data from normal livers and patients with early ALD and severe AH were analyzed. α-Diversity indices and percentage Shannon diversity of a gene, which refers to this gene’s contribution to total Shannon entropy, were calculated. Ingenuity pathway analysis identified canonical pathways determined by DEG and DSD approaches. ALD significantly decreased hepatic transcriptome α-diversity, correlating with increased relative contribution of select genes. These changes were driven by lower-abundance gene expression loss. DEG and DSD analyses showed overlapping genes and canonical pathways, but DSD also identified additional genes and pathways not highlighted by DEGs, including fatty acid oxidation, extracellular matrix degradation, and cholesterol metabolism pathways that may represent additional therapeutic targets. Importantly, DSD more effectively identified differences between ASH and AH. Overall, α-diversity analysis revealed that ALD progressively reduces transcriptome heterogeneity, and that DSD provides complementary insights into disease mechanisms missed by standard approaches.

The work represents a notable methodological advancement that bridges ecological theory with clinical hepatology, offering new perspectives on disease progression and potentially informative biomarkers.