Hiroki Kobayashi1, Krystyna A Gieniec2, Josephine A Wright3, Tongtong Wang2, Naoya Asai4, Yasuyuki Mizutani5, Tadashi Lida5, Ryota Ando6, Nobumi Suzuki7, Tamsin R M Lannagan2, Jia Q Ng2, Akitoshi Hara8, Yukihiro Shiraki6, Shinji Mii9, Mari Ichinose2, Laura Vrbanac2, Matthew J Lawrence10, Tarik Sammour11, Kay Uehara12, Gareth Davies13, Leszek Lisowski14, Ian E Alexander15, Yoku Hayakawa16, Lisa M Butler2, Andrew C W Zannettino2, M Omar Din17, Jeff Hasty18, Alastair D Burt19, Simon J Leedham20, Anil K Rustgi21, Siddhartha Mukherjee22, Timothy C Wang22, Atsushi Enomoto23, Masahide Takahashi24, Daniel L Worthley25, Susan L Woods26. 1. Adelaide Medical School, University of Adelaide, Adelaide, Australia; South Australian Health and Medical Research Institute, Adelaide, Australia; Department of Pathology, Nagoya University Graduate School of Medicine, Nagoya, Japan; Division of Molecular Pathology, Center for Neurological Disease and Cancer, Nagoya University Graduate School of Medicine, Nagoya, Japan. 2. Adelaide Medical School, University of Adelaide, Adelaide, Australia; South Australian Health and Medical Research Institute, Adelaide, Australia. 3. South Australian Health and Medical Research Institute, Adelaide, Australia. 4. Department of Molecular Pathology, Graduate School of Medicine, Fujita Health University, Toyoake, Japan. 5. Department of Pathology, Nagoya University Graduate School of Medicine, Nagoya, Japan; Department of Gastroenterology and Hepatology, Nagoya University Graduate School of Medicine, Nagoya, Japan. 6. Department of Pathology, Nagoya University Graduate School of Medicine, Nagoya, Japan. 7. Adelaide Medical School, University of Adelaide, Adelaide, Australia; South Australian Health and Medical Research Institute, Adelaide, Australia; Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan. 8. Department of Cardiology, Nagoya University Graduate School of Medicine, Nagoya, Japan. 9. Department of Pathology, Nagoya University Graduate School of Medicine, Nagoya, Japan; Division of Molecular Pathology, Center for Neurological Disease and Cancer, Nagoya University Graduate School of Medicine, Nagoya, Japan. 10. Colorectal Unit, Department of Surgery, Royal Adelaide Hospital, Adelaide, Australia. 11. Adelaide Medical School, University of Adelaide, Adelaide, Australia; South Australian Health and Medical Research Institute, Adelaide, Australia; Colorectal Unit, Department of Surgery, Royal Adelaide Hospital, Adelaide, Australia. 12. Division of Surgical Oncology, Department of Surgery, Nagoya University Graduate School of Medicine, Nagoya, Japan. 13. UCB Pharma, Slough, Berkshire, United Kingdom. 14. Translational Vectorology Research Unit, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia; Vector and Genome Engineering Facility, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Westmead, Australia; Military Institute of Hygiene and Epidemiology, The Biological Threats Identification and Countermeasure Centre, Puławy, Poland. 15. Gene Therapy Research Unit, Sydney Children's Hospitals Network and Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia; Discipline of Child and Adolescent Health, Faculty of Medicine and Health, The University of Sydney, Australia. 16. Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan. 17. GenCirq, Inc, San Diego, California. 18. Department of Bioengineering, University of California, San Diego, La Jolla, California. 19. Adelaide Medical School, University of Adelaide, Adelaide, Australia; Precision and Molecular Pathology, Newcastle University, Newcastle Upon Tyne, United Kingdom. 20. Intestinal Stem Cell Biology Lab, Wellcome Trust Centre Human Genetics, University of Oxford, Oxford, United Kingdom. 21. Herbert Irving Comprehensive Cancer Center, Division of Digestive and Liver Diseases, Department of Medicine, Columbia University, New York, New York. 22. Department of Medicine and Irving Cancer Research Center, Columbia University, New York, New York. 23. Department of Pathology, Nagoya University Graduate School of Medicine, Nagoya, Japan. Electronic address: enomoto@iar.nagoya-u.ac.jp. 24. Department of Pathology, Nagoya University Graduate School of Medicine, Nagoya, Japan; Division of Molecular Pathology, Center for Neurological Disease and Cancer, Nagoya University Graduate School of Medicine, Nagoya, Japan; International Center for Cell and Gene Therapy, Fujita Health University, Toyoake, Japan. Electronic address: mtakaha@med.nagoya-u.ac.jp. 25. South Australian Health and Medical Research Institute, Adelaide, Australia. Electronic address: Dan.Worthley@sahmri.com. 26. Adelaide Medical School, University of Adelaide, Adelaide, Australia; South Australian Health and Medical Research Institute, Adelaide, Australia. Electronic address: susan.woods@adelaide.edu.au.
Abstract
BACKGROUND & AIMS: Cancer-associated fibroblasts (CAFs), key constituents of the tumor microenvironment, either promote or restrain tumor growth. Attempts to therapeutically target CAFs have been hampered by our incomplete understanding of these functionally heterogeneous cells. Key growth factors in the intestinal epithelial niche, bone morphogenetic proteins (BMPs), also play a critical role in colorectal cancer (CRC) progression. However, the crucial proteins regulating stromal BMP balance and the potential application of BMP signaling to manage CRC remain largely unexplored. METHODS: Using human CRC RNA expression data, we identified CAF-specific factors involved in BMP signaling, then verified and characterized their expression in the CRC stroma by in situ hybridization. CRC tumoroids and a mouse model of CRC hepatic metastasis were used to test approaches to modify BMP signaling and treat CRC. RESULTS: We identified Grem1 and Islr as CAF-specific genes involved in BMP signaling. Functionally, GREM1 and ISLR acted to inhibit and promote BMP signaling, respectively. Grem1 and Islr marked distinct fibroblast subpopulations and were differentially regulated by transforming growth factor β and FOXL1, providing an underlying mechanism to explain fibroblast biological dichotomy. In patients with CRC, high GREM1 and ISLR expression levels were associated with poor and favorable survival, respectively. A GREM1-neutralizing antibody or fibroblast Islr overexpression reduced CRC tumoroid growth and promoted Lgr5+ intestinal stem cell differentiation. Finally, adeno-associated virus 8 (AAV8)-mediated delivery of Islr to hepatocytes increased BMP signaling and improved survival in our mouse model of hepatic metastasis. CONCLUSIONS: Stromal BMP signaling predicts and modifies CRC progression and survival, and it can be therapeutically targeted by novel AAV-directed gene delivery to the liver.
BACKGROUND & AIMS: Cancer-associated fibroblasts (CAFs), key constituents of the tumor microenvironment, either promote or restrain tumor growth. Attempts to therapeutically target CAFs have been hampered by our incomplete understanding of these functionally heterogeneous cells. Key growth factors in the intestinal epithelial niche, bone morphogenetic proteins (BMPs), also play a critical role in colorectal cancer (CRC) progression. However, the crucial proteins regulating stromal BMP balance and the potential application of BMP signaling to manage CRC remain largely unexplored. METHODS: Using human CRC RNA expression data, we identified CAF-specific factors involved in BMP signaling, then verified and characterized their expression in the CRC stroma by in situ hybridization. CRC tumoroids and a mouse model of CRC hepatic metastasis were used to test approaches to modify BMP signaling and treat CRC. RESULTS: We identified Grem1 and Islr as CAF-specific genes involved in BMP signaling. Functionally, GREM1 and ISLR acted to inhibit and promote BMP signaling, respectively. Grem1 and Islr marked distinct fibroblast subpopulations and were differentially regulated by transforming growth factor β and FOXL1, providing an underlying mechanism to explain fibroblast biological dichotomy. In patients with CRC, high GREM1 and ISLR expression levels were associated with poor and favorable survival, respectively. A GREM1-neutralizing antibody or fibroblast Islr overexpression reduced CRC tumoroid growth and promoted Lgr5+ intestinal stem cell differentiation. Finally, adeno-associated virus 8 (AAV8)-mediated delivery of Islr to hepatocytes increased BMP signaling and improved survival in our mouse model of hepatic metastasis. CONCLUSIONS: Stromal BMP signaling predicts and modifies CRC progression and survival, and it can be therapeutically targeted by novel AAV-directed gene delivery to the liver.
Authors: Hiroki Kobayashi; Krystyna A Gieniec; Tamsin R M Lannagan; Tongtong Wang; Naoya Asai; Yasuyuki Mizutani; Tadashi Iida; Ryota Ando; Elaine M Thomas; Akihiro Sakai; Nobumi Suzuki; Mari Ichinose; Josephine A Wright; Laura Vrbanac; Jia Q Ng; Jarrad Goyne; Georgette Radford; Matthew J Lawrence; Tarik Sammour; Yoku Hayakawa; Sonja Klebe; Alice E Shin; Samuel Asfaha; Mark L Bettington; Florian Rieder; Nicholas Arpaia; Tal Danino; Lisa M Butler; Alastair D Burt; Simon J Leedham; Anil K Rustgi; Siddhartha Mukherjee; Masahide Takahashi; Timothy C Wang; Atsushi Enomoto; Susan L Woods; Daniel L Worthley Journal: Gastroenterology Date: 2021-12-06 Impact factor: 33.883