Living Gut Bacteria Functionalized with Gold Nanoclusters and Drug for Facile Cancer Theranostics

Bacbots are potent self-propelling vehicles for targeted therapy that can be guided by chemical and biochemical stimuli of the host. In addition, they can be guided externally by the use of magnetic field or other physical forces. The challenge is to incorporate drugs and diagnostic tools in living bacteria with retention of theranostic activity until reaching the targets and easy clearance of the remainder following the treatment. We report that living Lactobacillus rhamnosus, when functionalized with photoluminescent Au nanoclusters and the anticancer drug methotrexate, was cytotoxic to monolayer and spheroids of cancer cells (HeLa and HT29) even at a low dose of bacteria used (10⁷ cfu/mL). The observed cell death was nearly 90% in HeLa spheroids and 70% in HT29 spheroids. Further, functionalization of the bacterial surface with the nanoclusters helped incorporate the drug onto their cell surfaces. The drug and nanocluster-loaded bacteria annihilated the cells and the spheroids in a rather short time (6 h) that revealed the specificity and effectiveness of the bacbots. The bacbots exhibited synergistic toxicity on the cells as their effect was more than the drug and the bacteria individually. This higher toxicity could be associated with elevated levels of reactive oxygen species generated in the bacbot-treated cells. The multifunctional bacbots reported here provide an option for guided therapy with the natural variant of the human gut-friendly living bacteria without the need for attenuation or genetic modification.