Organoid modelling identifies that DACH1 functions as a tumour promoter in colorectal cancer by modulating BMP signalling

EBioMedicine

Weixing Dai¹  Renjie Wang²  Qiang Guo²  Sanjun Cai²  Xinxin Rao²  Xiaoji Ma²  Long Zhang²  Zhen Zhang³  Xiaoxue Gao³  Yaqi Li³  Mengxue Pan³  Guoxiang Fu³  Xiang Hu³  Yi Zhou⁴  Xiaoya Xu⁴  Jianjun Gao⁴  Junjie Peng⁴  Guoqiang Hua⁴ 

[1] Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China;Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China;Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China;Institute of Radiation Medicine, and Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China;

关键词: DACH1;    Organoid;    Stem cells;    BMP signalling pathway;    Colorectal cancer;   

DOI  :  

来源: DOAJ

【 摘 要 】

Background: Dachshund homologue 1 (DACH1) is highly expressed in LGR5+ intestinal stem cells and colorectal tumours. However, the roles of DACH1 in intestinal cell stemness and colorectal tumorigenesis remain largely undefined. Methods: We used immunohistochemistry, western blotting and quantitative real-time PCR to analyse DACH1 expression in colorectal cancer (CRC) samples. CRISPR/Cas9 gene editing and lentiviral vector-mediated overexpression and shRNA-mediated knockdown of DACH1 were utilized to modulate DACH1 expression in cell lines and organoids. An intestinal organoid-based functional model was analysed, and cancer cell colony formation, sphere formation assays and murine xenotransplants were performed to reveal the role of DACH1 in CRC cell proliferation, stemness and tumorigenesis. Immunofluorescence, co-immunoprecipitation, RNA interference and microarray data analyses were conducted to demonstrate the association between DACH1 and the bone morphogenetic protein (BMP) signalling pathway. Findings: DACH1 is specifically expressed in discrete crypt base cells, and increased DACH1 expression was found in all stages of CRC. Moreover, the high expression of DACH1 independently predicted poor prognosis. In colon cancer cells, shRNA-mediated suppression of DACH1 inhibited cell growth in vitro and in vivo. By studying the intestinal organoid-based functional model, we found that depletion of DACH1 reduced the organoid formation efficiency and tumour organoid size. DACH1 overexpression stimulated both colonsphere formation and tumour organoid formation in the context of dysregulated BMP signalling. Mechanistic characterizations indicated that overexpression of DACH1 affects a subset of stem cell signature genes implicated in stem cell proliferation and maintenance through the suppression of BMP signalling via SMAD4. Interpretation: Together, our study highlights DACH1 as an integral regulator of BMP signalling during intestinal tumorigenesis, and DACH1 could be a potential prognostic marker and therapeutic target for colorectal cancer patients.

【 授权许可】

Unknown