Investigation of Exome-Wide Tumor Heterogeneity on Colorectal Tissue-Based Single Cells

Area of Science:

• Genomics and Molecular Biology
• Oncology
• Cancer Research

Background:

• Colorectal cancer progression is significantly impacted by environmental and genetic factors.
• Tumor heterogeneity is a key driver, but its single-cell level effects remain understudied.
• Current methods like cfDNA and bulk sequencing have limitations in capturing single-cell resolution.

Purpose of the Study:

• To develop and apply an end-to-end single-cell sequencing workflow for colorectal cancer.
• To investigate tumor heterogeneity patterns at the single-cell level using laser microdissection.
• To explore tissue-specific heterogeneity motifs across healthy, tumor-adjacent normal, and cancerous tissues.

Main Methods:

• Developed a comprehensive single-cell sequencing workflow from tissue sample isolation to exome sequencing.
• Utilized laser microdissection to isolate samples from distinct regions of tissue slides.
• Compared the performance of single-cell, whole-exome bulk, and cfDNA sequencing for single nucleotide variation analysis.

Main Results:

• Successfully implemented a single-cell sequencing workflow for detailed analysis of colorectal cancer.
• Identified and characterized distinct heterogeneity patterns within colorectal tumors at the single-cell level.
• Revealed tissue-specific heterogeneity motifs by analyzing healthy, tumor-associated normal, and cancer tissues.

Conclusions:

• Single-cell sequencing provides crucial insights into colorectal cancer heterogeneity previously unattainable.
• The developed workflow enables precise mapping of tumor heterogeneity and tissue-specific variations.
• This approach offers a more comprehensive understanding of colorectal cancer evolution and progression.