Chemoprevention—the use of medication to prevent cancer—can be augmented by the consumption of produce enriched with natural metabolites. However, chemopreventive metabolites are typically inactive and have low bioavailability and poor host absorption. Here, we show that engineered commensal microbes can prevent carcinogenesis and promote the regression of colorectal cancer through a cruciferous vegetable diet. The engineered commensal Escherichia coli bound specifically to the heparan sulphate proteoglycan on colorectal cancer cells and secreted the enzyme myrosinase to transform host-ingested glucosinolates—natural components of cruciferous vegetables—to sulphoraphane, an organic small molecule with known anticancer activity. The engineered microbes coupled with glucosinolates resulted in >95% proliferation inhibition of murine, human and colorectal adenocarcinoma cell lines in vitro. We also show that murine models of colorectal carcinoma fed with the engineered microbes and the cruciferous vegetable diet displayed significant tumour regression and reduced tumour occurrence.
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Howlander, N. et al. SEER Cancer Statistics Review, 1975–2011 (National Cancer Institute, Bethesda, MD, 2014).
Wollowski, I., Rechkemmer, G. & Pool-Zobel, B. L. Protective role of probiotics and prebiotics in colon cancer. Am. J. Clin. Nutr. 73, 451S–455S (2001).
Pietinen, P. et al. Diet and risk of colorectal cancer in a cohort of Finnish men. Cancer Causes Control 10, 387–396 (1999).
Liong, M.-T. Roles of probiotics and prebiotics in colon cancer prevention: postulated mechanisms and in-vivo evidence. Int. J. Mol. Sci. 9, 854–863 (2008).
American Cancer Society Treatment of Colon Cancer, by Stage (2016); http://www.cancer.org/cancer/colonandrectumcancer/detailedguide/colorectal-cancer-treating-by-stage-colon#
Donaldson, M. S. Nutrition and cancer: a review of the evidence for an anti-cancer diet. Nutr. J. 3, 1–21 (2004).
Divisi, D., Di Tommaso, S., Salvemini, S., Garramone, M. & Crisci, R. Diet and cancer. Acta Biomed. 77, 118–123 (2006).
Tortorella, S. M., Royce, S. G., Licciardi, P. V. & Karagiannis, T. C. Dietary sulforaphane in cancer chemoprevention: the role of epigenetic regulation and HDAC inhibition. Antioxid. Redox Signal. 22, 1382–1424 (2015).
Devi, J. R. & Thangam, E. B. Mechanisms of anticancer activity of sulforaphane from Brassica oleracea in HEp-2 human epithelial carcinoma cell line. Asian Pac. J. Cancer Prev. 13, 2095–2100 (2012).
Qazi, A. et al. Anticancer activity of a broccoli derivative, sulforaphane, in barrett adenocarcinoma: potential use in chemoprevention and as adjuvant in chemotherapy. Transl. Oncol. 3, 389–399 (2010).
Cramer, J. M. & Jeffery, E. H. Sulforaphane absorption and excretion following ingestion of a semi-purified broccoli powder rich in glucoraphanin and broccoli sprouts in healthy men. Nutr. Cancer 63, 196–201 (2011).
Li, Y. & Zhang, T. Targeting cancer stem cells with sulforaphane, a dietary component from broccoli and broccoli sprouts. Future Oncol. 9, 1097–1103 (2013).
Saeidi, N. et al. Engineering microbes to sense and eradicate Pseudomonas aeruginosa, a human pathogen. Mol. Syst. Biol. 7, 521 (2011).
Hwang, I. Y. et al. Reprogramming microbes to be pathogen-seeking killers. ACS Synt. Biol. 3, 228–237 (2014).
Duan, F. & March, J. C. Engineered bacterial communication prevents Vibrio cholerae virulence in an infant mouse model. Proc. Natl Acad. Sci. USA 107, 11260–11264 (2010).
Duan, F. F., Liu, J. H. & March, J. C. Engineered commensal bacteria reprogram intestinal cells into glucose-responsive insulin-secreting cells for the treatment of diabetes. Diabetes 64, 1794–1803 (2015).
Ho, C. L., Hwang, I. Y., Loh, K. & Chang, M. W. Matrix-immobilized yeast for large-scale production of recombinant human lactoferrin. MedChemComm 6, 486–491 (2015).
Danino, T. et al. Programmable probiotics for detection of cancer in urine. Sci. Transl. Med. 7, 289ra84 (2015).
Wu, H. C. et al. Autonomous bacterial localization and gene expression based on nearby cell receptor density. Mol. Syst. Biol. 9, 636 (2013).
Frahm, M. et al. Efficiency of conditionally attenuated Salmonella enterica serovar typhimurium in bacterium-mediated tumor therapy. MBio 6, e00254 (2015).
Pye, G., Evans, D. F., Ledingham, S. & Hardcastle, J. D. Gastrointestinal intraluminal pH in normal subjects and those with colorectal adenoma or carcinoma. Gut 31, 1355–1357 (1990).
Singh, N. R., Denissen, E. C., McKune, A. J. & Peters, E. M. Intestinal temperature, heart rate, and hydration status in multiday trail runners. Clin. J. Sport Med. 22, 311–318 (2012).
Gerlt, J. A. et al. Enzyme Function Initiative-Enzyme Similarity Tool (EFI-EST): a web tool for generating protein sequence similarity networks. Biochim. Biophys. Acta 1854, 1019–1037 (2015).
Burmeister, W. P. et al. The crystal structures of Sinapis alba myrosinase and a covalent glycosyl-enzyme intermediate provide insights into the substrate recognition and active-site machinery of an S-glycosidase. Structure 5, 663–675 (1997).
Naushad, M., Alothman, Z. A., Khan, A. B. & Ali, M. Effect of ionic liquid on activity, stability, and structure of enzymes: a review. Int. J. Biol. Macromol. 51, 555–560 (2012).
Strickler, S. S. et al. Protein stability and surface electrostatics: a charged relationship. Biochemistry 45, 2761–2766 (2006).
Botti, M. G., Taylor, M. G. & Botting, N. P. Studies on the mechanism of myrosinase. Investigation of the effect of glycosyl acceptors on enzyme activity. J. Biol. Chem. 270, 20530–20535 (1995).
Zhang, Y., Tang, L. & Gonzalez, V. Selected isothiocyanates rapidly induce growth inhibition of cancer cells. Mol. Cancer Ther. 2, 1045–1052 (2003).
Lai, K. et al. Allyl isothiocyanate inhibits cell metastasis through suppression of the MAPK pathways in epidermal growth factor-stimulated HT29 human colorectal adenocarcinoma cells. Oncol. Rep. 31, 189–196 (2014).
Lau, W. S., Chen, T. & Wong, Y. S. Allyl isothiocyanate induces G2/M arrest in human colorectal adenocarcinoma SW620 cells through down-regulation of Cdc25B and Cdc25C. Mol. Med. Rep. 3, 1023–1030 (2010).
Wagner, A. E., Boesch-Saadatmandi, C., Dose, J., Schultheiss, G. & Rimbach, G. Anti-inflammatory potential of allyl-isothiocyanate—role of Nrf2, NF-κB and microRNA-155. J. Cell. Mol. Med. 16, 836–843 (2012).
Cheng, B. et al. Syndecan as cell surface receptors in cancer biology. A focus on their interaction with PDZ domain proteins. Front. Pharmacol. 7, 10 (2016).
Sanderson, R. D. et al. Enzymatic remodeling of heparan sulfate proteoglycans within the tumor microenvironment: growth regulation and the prospect of new cancer therapies. J. Cell. Biochem. 96, 897–905 (2005).
Rosen, S. D. & Lemjabbar-Alaoui, H. Sulf-2: an extracellular modulator of cell signaling and a cancer target candidate. Expert Opin. Ther. Targets 14, 935–949 (2010).
Li, Q. et al. Molecular characterization of an ice nucleation protein variant (InaQ) from Pseudomonas syringae and the analysis of its transmembrane transport activity in Escherichia coli. Int. J. Biol. Sci. 8, 1097–1108 (2012).
Neufert, C., Becker, C. & Neurath, M. F. An inducible mouse model of colon carcinogenesis for the analysis of sporadic and inflammation-driven tumor progression. Nat. Protoc. 2, 1998–2004 (2007).
De Robertis, M. et al. The AOM/DSS murine model for the study of colon carcinogenesis: from pathways to diagnosis and therapy studies. J. Carcinog. 10, 9 (2011).
Haggar, F. A. & Boushey, R. P. Colorectal cancer epidemiology: incidence, mortality, survival, and risk factors. Clin. Colon Rectal Surg. 22, 191–197 (2009).
Schultz, M. Clinical use of E. coli Nissle 1917 in inflammatory bowel disease. Inflamm. Bowel Dis. 14, 1012–1018 (2008).
Newman, D. J. & Cragg, G. M. Microbial antitumor drugs: natural products of microbial origin as anticancer agents. Curr. Opin. Investig. Drugs 10, 1280–1296 (2009).
Mathonnet, M. et al. Hallmarks in colorectal cancer: angiogenesis and cancer stem-like cells. World J. Gastroenterol. 20, 4189–4196 (2014).
Byrd, J. C. & Bresalier, R. S. Mucins and mucin binding proteins in colorectal cancer. Cancer Metastasis Rev. 23, 77–99 (2004).
Kuppusamy, P., Govindan, N., Yusoff, M. M. & Ichwan, S. J. A. Proteins are potent biomarkers to detect colon cancer progression. Saudi J. Biol. Sci. 24, 1212–1221 (2017).
Boleij, A. et al. Surface-exposed histone-like protein a modulates adherence of Streptococcus gallolyticus to colon adenocarcinoma cells. Infect. Immun. 77, 5519–5527 (2009).
American Cancer Society Cancer Facts & Figures 2015 (2015); http://www.cancer.org/research/cancerfactsstatistics/cancerfactsfigures2015/
Dienstmann, R., Salazar, R. & Tabernero, J. Personalizing colon cancer adjuvant therapy: selecting optimal treatments for individual patients. J. Clin. Oncol. 33, 1787–1796 (2015).
Pai, S. G. & Fuloria, J. Novel therapeutic agents in the treatment of metastatic colorectal cancer. World J. Gastrointest. Oncol. 8, 99–104 (2016).
St Jean, A. T., Swofford, C. A., Panteli, J. T., Brentzel, Z. J. & Forbes, N. S. Bacterial delivery of Staphylococcus aureus α-hemolysin causes regression and necrosis in murine tumors. Mol. Ther. 22, 1266–1274 (2014).
Nemunaitis, J. et al. Pilot trial of genetically modified, attenuated Salmonella expressing the E. coli cytosine deaminase gene in refractory cancer patients. Cancer Gene Ther. 10, 737–744 (2003).
Teicher, B. A. & Andrews, P. A. Anticancer Drug Development Guide: Preclinical Screening, Clinical Trials, and Approval Vol. 2 (Humana, New York, USA, 2004).
We thank P. Han for comments on the manuscript. This work was supported by the Agency for Science, Technology and Research (A*STAR) of Singapore (112 177 0040), Synthetic Biology Initiative of the National University of Singapore (DPRT/943/09/14), Summit Research Program of the National University Health System (NUHSRO/2016/053/SRP/05) and US Defense Threat Reduction Agency (HDTRA1-13-0037). We recognize the administrative contributions of C. Chang, I. Y. Hwang, H. L. Pham, B. E. D. Buenaflor, S. J. David and A. Seok Ting to the pre-clinical study.
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Nature Chemical Biology (2019)
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