Mapping and tracing Grem1+ stromal cells in an Apc Min/+ mouse utilizing cryopreserved intestinal sections prepared via modified Swiss-roll technique

Youheng Jiang; Zhang Fu; Yanfang Chen; Qunlong Jin; Yanming Yang; Zerong Lin; Changxue Li; Yunfei Gao; Zepeng Dong; Yang He; Xinjun Mao; Yulong He; Qingyuan Zhang; Qi Zhang; Ningning Li

doi:10.1016/j.isci.2024.111173

Mapping and tracing Grem1⁺ stromal cells in an Apc ^Min/+ mouse utilizing cryopreserved intestinal sections prepared via modified Swiss-roll technique

iScience. 2024 Oct 15;27(11):111173. doi: 10.1016/j.isci.2024.111173. eCollection 2024 Nov 15.

Authors

Youheng Jiang^{1

2}, Zhang Fu^{1

3}, Yanfang Chen², Qunlong Jin^{1

2}, Yanming Yang¹, Zerong Lin¹, Changxue Li^{1

2}, Yunfei Gao^{1

4}, Zepeng Dong¹, Yang He⁵, Xinjun Mao⁶, Yulong He^{1

2}, Qingyuan Zhang², Qi Zhang⁷, Ningning Li^{1

8}

Affiliations

¹ Tomas Lindahl Nobel Laureate Laboratory, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen 518107, China.
² Digestive Diseases Center, Guangdong Provincial Key Laboratory of Digestive Cancer Research, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen 518107, China.
³ Department of Geriatrics, The Seventh Affiliated Hospital of Sun Yat-Sen University, Shenzhen 518107, China.
⁴ Department of Otolaryngology, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen 518107, China.
⁵ School of Mechanical Engineering and Automation, Harbin Institute of Technology, Shenzhen 518107, China.
⁶ Department of Anesthesiology, The Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000, China.
⁷ Department of Anesthesiology, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen 518107, China.
⁸ China-UK Institute for Frontier Science, Shenzhen 518107, China.

Abstract

Grem1⁺ cancer-associated fibroblasts (CAFs) are crucial in colorectal cancer (CRC) development, yet technical challenges have limited understanding of their origins, spatiotemporal distribution, and potential roles. Here, we devised a custom mold, optimizing the gut Swiss-roll technique to create a single cryopreserved slide for comprehensive staining. Our integrated approach uncovered a marked increase in Grem1⁺ CAFs within Apc ^Min/+ mouse tumors at 12 weeks, compared to normal mucosa. Subsequent lineage tracing in Grem1-CreER ^T2 ; R26-LSL-tdTomato; Apc ^Min/+ mice revealed that most Grem1⁺ CAFs infiltrating the tumor core originated from Grem1⁺ intestinal reticular stem cells (iRSCs). A minor subset of Grem1⁺ CAFs, located in the submucosa, retained characteristics of Grem1⁺ intestinal sub-epithelial myofibroblasts (ISEMFs). Altogether, CAFs derived from Grem1⁺ iRSCs may serve as a principal stromal cell type driving early-stage CRC progression, while Grem1⁺ ISEMFs contribute less from a more distant location. Hence, targeting Grem1⁺ CAFs presents an early and promising therapeutic strategy in CRC.

Keywords: Cancer; Cell biology; Molecular biology.