title: Programmable probiotics for detection of cancer in urine.
abstract: Rapid advances in the forward engineering of genetic circuitry in living cells has positioned synthetic biology as a potential means to solve numerous biomedical problems, including disease diagnosis and therapy. One challenge in exploiting synthetic biology for translational applications is to engineer microbes that are well tolerated by patients and seamlessly integrate with existing clinical methods. We use the safe and widely used probiotic Escherichia coli Nissle 1917 to develop an orally administered diagnostic that can noninvasively indicate the presence of liver metastasis by producing easily detectable signals in urine. Our microbial diagnostic generated a high-contrast urine signal through selective expansion in liver metastases (10(6)-fold enrichment) and high expression of a lacZ reporter maintained by engineering a stable plasmid system. The lacZ reporter cleaves a substrate to produce a small molecule that can be detected in urine. E. coli Nissle 1917 robustly colonized tumor tissue in rodent models of liver metastasis after oral delivery but did not colonize healthy organs or fibrotic liver tissue. We saw no deleterious health effects on the mice for more than 12 months after oral delivery. Our results demonstrate that probiotics can be programmed to safely and selectively deliver synthetic gene circuits to diseased tissue microenvironments in vivo.
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA.
authors.forename: Tal
authors.initials: T
authors.lastname: Danino
authors.affiliation: Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093, USA.
authors.forename: Arthur
authors.initials: A
authors.lastname: Prindle
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA.
authors.forename: Gabriel A
authors.initials: GA
authors.lastname: Kwong
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Skalak
authors.affiliation: Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093, USA.
authors.forename: Howard
authors.initials: H
authors.lastname: Li
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA.
authors.forename: Kaitlin
authors.initials: K
authors.lastname: Allen
authors.affiliation: Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093, USA. BioCircuits Institute, University of California, San Diego, La Jolla, CA 92093, USA. Molecular Biology Section, Division of Biological Science, University of California, San Diego, La Jolla, CA 92093, USA.
authors.forename: Jeff
authors.initials: J
authors.lastname: Hasty
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA. Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA. Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA. Electrical Engineering and Computer Science and David H. Koch Institute for Integrative Cancer Research, MIT, Cambridge, MA 02139, USA. Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA. sbhatia@mit.edu.
authors.forename: Sangeeta N
authors.initials: SN
authors.lastname: Bhatia
mesh_terms: D000284:Administration, Oral; D000818:Animals; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D055088:Early Detection of Cancer / Q000379:methods*; D006801:Humans; D008113:Liver Neoplasms / Q000382:microbiology / Q000556:secondary; D008819:Mice, Nude; D009369:Neoplasms / Q000175:diagnosis* / Q000652:urine*; D010957:Plasmids / Q000378:metabolism; D019936:Probiotics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013997:Time Factors; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D001616:beta-Galactosidase