Enhanced Spatial Transcriptomics Analysis of Mouse Lung Tissues Reveals Cell-Specific Gene Expression Changes...

Area of Science:

• Molecular Biology
• Genomics
• Pathology

Background:

• Spatial transcriptomics preserves tissue context lost in traditional methods.
• Understanding cellular changes in pulmonary hypertension (PH) requires spatial information.

Purpose of the Study:

• To develop and validate a high-resolution spatial transcriptomics protocol for fixed frozen mouse lung sections.
• To analyze cellular and gene expression changes in a mouse model of PH.

Main Methods:

• Utilized 10X Genomics Xenium platform with multiplexed fluorescent in situ hybridization (FISH).
• Compared fixed frozen and fresh frozen tissue preparation methods.
• Analyzed gene expression using a 379-gene mouse panel on fixed frozen lung sections.

Main Results:

• The fixed frozen protocol preserved tissue integrity and provided high-fidelity spatial transcriptomics data.
• Identified 40 major lung cell types and detected alterations in PH, including increased arterial endothelial cells and fibroblasts.
• Observed markers of endothelial-to-mesenchymal transition and fibroblast activation, confirming increased arterialization in PH lungs.

Conclusions:

• The developed protocol reliably integrates spatial and transcriptomic data from fixed frozen lung tissues.
• Spatial transcriptomics is crucial for understanding cellular heterogeneity and disease mechanisms in complex conditions like PH.