ABSTRACT
Immunotherapeutic approaches to manage patients with advanced gastrointestinal malignancies are desired; however, mechanisms to incite tumor-specific immune responses remain to be elucidated. Rose bengal (RB) is toxic at low concentrations to malignant cells and may induce damage-associated molecular patterns; therefore, we investigated its potential as an immunomodulator in colon cancer. Murine and human colon cancer lines were treated with RB (10% in saline/PV-10) for cell cycle, cell death, and apoptosis assays. Damage-associated molecular patterns were assessed with western blot, ELISA, and flow cytometry. In an immunocompetent murine model of colon cancer, we demonstrate that tumors regress upon RB treatment, and that RB induces cell death in colon cancer cells through G2/M growth arrest and predominantly necrosis. RB-treated colon cancer cells expressed distinct hallmarks of immunogenic cell death (ICD), including enhanced expression of calreticulin and heat-shock protein 90 on the cell surface, a decrease in intracellular ATP, and the release of HMGB1. To confirm the ICD phenotype, we vaccinated immunocompetent animals with syngeneic colon cancer cells treated with RB. RB-treated tumors served as a vaccine against subsequent challenge with the same CT26 colon cancer tumor cells, and vaccination with in vitro RB-treated cells resulted in slower tumor growth following inoculation with colon cancer cells, but not with syngeneic non-CT26 cancer cells, suggesting a specific antitumor immune response. In conclusion, RB serves as an inducer of ICD that contributes to enhanced specific antitumor immunity in colorectal cancer.
Subject(s)
Antineoplastic Agents/immunology , Antineoplastic Agents/pharmacology , Cell Death/drug effects , Cell Death/immunology , Colonic Neoplasms/drug therapy , Colonic Neoplasms/immunology , Rose Bengal/pharmacology , Animals , Apoptosis/drug effects , Apoptosis/immunology , Calreticulin/metabolism , Cell Line, Tumor , Colonic Neoplasms/metabolism , HCT116 Cells , HMGB1 Protein/metabolism , HSP90 Heat-Shock Proteins/metabolism , HT29 Cells , Humans , Mice , Mice, Inbred BALB C , Necrosis/drug therapy , Necrosis/immunology , Necrosis/metabolismSubject(s)
Duodenal Neoplasms/surgery , Gastrointestinal Stromal Tumors/surgery , Pancreas/abnormalities , Pancreatic Diseases/diagnosis , Pancreatic Diseases/surgery , Aged, 80 and over , Duodenal Neoplasms/diagnosis , Endosonography/methods , Female , Gastrointestinal Hemorrhage/diagnosis , Gastrointestinal Hemorrhage/etiology , Gastrointestinal Stromal Tumors/diagnosis , Humans , Intraoperative Care/methods , Pancreas/surgery , Pancreaticoduodenectomy/methods , Preoperative Care/methods , Rare Diseases , Rectum , Risk Assessment , Tomography, X-Ray Computed/methods , Treatment OutcomeABSTRACT
Rose Bengal (RB) is a red synthetic dye that was initially used in the garment industry and has been used safely for decades as a corneal stain by ophthalmologists. Antineoplastic properties of RB have also been observed, though the mechanism of action remained to be elucidated. Recently, interest in RB as a therapeutic cancer treatment has increased due to significant anti-tumor responses with direct tumor injection in human clinical trials for metastatic melanoma. In these patients, there has been the implication that RB may mount a T-cell mediated anti-tumor response and impart antigen-specific responses in distant bystander lesions. This article serves to evaluate the potential of intralesional rose bengal to stimulate T-cell mediated anti-tumor responses in in-vitro, pre-clinical, and clinical studies.
