Organoid modelling identifies that DACH1 functions as a tumour promoter in colorectal cancer by modulating BMP signalling
Xiang Hu,
Long Zhang,
Yaqi Li,
Xiaoji Ma,
Weixing Dai,
Xiaoxue Gao,
Xinxin Rao,
Guoxiang Fu,
Renjie Wang,
Mengxue Pan,
Qiang Guo,
Xiaoya Xu,
Yi Zhou,
Jianjun Gao,
Zhen Zhang,
Sanjun Cai,
Junjie Peng,
Guoqiang Hua
Affiliations
Xiang Hu
Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China
Long Zhang
Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China; Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China
Yaqi Li
Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China
Xiaoji Ma
Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China
Weixing Dai
Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China
Xiaoxue Gao
Institute of Radiation Medicine, and Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China
Xinxin Rao
Institute of Radiation Medicine, and Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China
Guoxiang Fu
Institute of Radiation Medicine, and Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China
Renjie Wang
Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China
Mengxue Pan
Institute of Radiation Medicine, and Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China
Qiang Guo
Institute of Radiation Medicine, and Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China
Xiaoya Xu
Institute of Radiation Medicine, and Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China
Yi Zhou
Institute of Radiation Medicine, and Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China
Jianjun Gao
Institute of Radiation Medicine, and Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China
Zhen Zhang
Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China; Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China
Sanjun Cai
Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China; Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China; Correspondence to: Sanjun Cai, Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Director's Office, 11th floor, building 3, No. 270, Dong'an Road, Xuhui District, Shanghai, 200032, China
Junjie Peng
Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China; Correspondence to: Junjie Peng, Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Director's Office, 11th floor, building 3, No. 270, Dong'an Road, Xuhui District, Shanghai, 200032, China
Guoqiang Hua
Institute of Radiation Medicine, and Cancer institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai 200032, China; Correspondence to: Guoqiang Hua, Institute of Radiation Medicine, Fudan University, No. 2094, Xietu Road, Xuhui District, Shanghai, 200032, China
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.
