hum0402-v1Release info

Elucidation of the mechanisms that give rise to driver mutations in colorectal carcinogenesis

Aims

Recently, single cell RNA sequencing has been used to analyze cell-cell interactions between colon cancer cells and stromal cells. However, the important cancer microenvironment during early cancer development from adenomas, especially the mechanism of immune tolerance acquisition, is unclear. In this study, we investigated immune tolerance and the formation of the tumor microenvironment at the adenoma-cancer interface by integrating single-cell and spatial transcriptome analyses obtained from public databases. By integrating scRNA-seq of colorectal cancer and spatial transcriptome analysis in carcinoma in adenoma tissues, we investigated immune tolerance and tumor microenvironment formation at the adenoma-tumor interface thereby searching for new therapeutic target molecules.

Methods

Formalin-fixed paraffin-embedded (FFPE) samples were used to prepare for spatial transcriptomic construction and sequencing. 5μm thick sections were prepared from the FFPE samples and processed using Visium Spatial Gene Expression Slide Kit (10x Genomics) according to the Visium Spatial Gene Expression Reagent Kits for FFPE User Guide. First, sections were stained with H&E and imaged followed by probe hybridization and ligation. Then, the captured probe library was sequenced using an MGI DNBSEQ-G400 system for 28, 10, 10, and 50 cycles for Read 1, i7, i5, and Read2 sequences respectively.

Participants/materials

Patients with colorectal cancer with a background of adenocarcinoma sequence (ACS)

URL

N/A

Principal investigator

Koshi Mimori

Affiliation

Department of Surgery, Kyushu University Beppu Hospital