Correlative multimodal imaging for microscale spatial mapping of collagen-gene activity interactions in human tissues

Area of Science:

• Biomedical imaging
• Molecular biology
• Tissue engineering

Background:

• Gene activity and tissue structure are crucial for understanding diseases like Duchenne Muscular Dystrophy.
• RNAscope offers single-molecule transcript detection, and multiphoton microscopy provides high-resolution collagen imaging.
• Integrating these techniques allows for spatial correlation of molecular and structural data.

Purpose of the Study:

• To develop and apply a correlative multimodal imaging workflow integrating RNAscope and Second Harmonic Generation microscopy.
• To investigate the spatial relationship between dystrophin transcript localization and collagen architecture in human skeletal muscle.
• To establish a versatile platform for spatial biomarker discovery and tissue analysis.

Main Methods:

• Correlative multimodal imaging combining RNAscope in situ hybridization with Second Harmonic Generation microscopy.
• High-resolution, quantitative imaging of extracellular matrix collagen and transcript localization within the same tissue section.
• Application to human skeletal muscle biopsies from healthy and Duchenne Muscular Dystrophy patients.

Main Results:

• Successfully integrated RNAscope and Second Harmonic Generation microscopy for microscale spatial correlation of molecular and structural data.
• Observed that regions enriched in specific dystrophin transcripts (exons 37-42 and 63-75) were associated with increased collagen fiber length and density.
• Demonstrated a potential spatial correlation between dystrophin transcript distribution and collagen organization in Duchenne Muscular Dystrophy muscle.

Conclusions:

• The developed workflow enables microscale integration of molecular and structural data in diverse tissues.
• This approach facilitates spatial biomarker discovery, fibrosis and regeneration studies, and evaluation of transcript-based therapies.
• The findings suggest a link between dystrophin transcript patterns and collagen organization in Duchenne Muscular Dystrophy, opening avenues for further research.