Mitigation of Ionizing Irradiation-Induced Intestinal Damage by Second-Generation Probiotics LR-IL-22 and LR-IFN-β

Abstract We have developed genetically modified second-generation probiotics for the localized delivery of known mitigators to the GI tract in order to reduce damage and regenerate tissue after exposure to ionizing radiation. Mitigating the effects of ionizing radiation exposure is critical for improving survival in the event of a radiological or nuclear (RAD-NUC) incident, where exposure could lead to hematopoietic or gastrointestinal (GI) acute radiation syndrome (ARS). While Neupogen and Neulasta have been granted label extensions by the FDA to treat casualties of a RAD-NUC incident, these drugs do not prevent GI-ARS-related mortality. There currently are no FDA-approved mitigators for GI-ARS. To address this critical need, we have engineered Lactobacillus reuteri to produce therapeutic cytokines (IL-22 or INF-β) and target these mitigators to the small intestines to recover intestinal stem cells, regenerate the radiation-sensitive intestinal crypts, and dramatically improve survival from 0–10% to 70–80% after exposure to GI-ARS-inducing radiation doses. This is a new strategy for therapeutic drug delivery, using a probiotic can be administered orally, facilitating its use in the context of a resource-limited mass casualty scenario. In addition to mitigating injury following a RAD-NUC incident, this approach is also applicable to radioprotection of the intestine during abdominal radiotherapy. Symptoms of GI toxicity affect 60–80% of the andgt;300,000 patients that receive pelvic or abdominal radiation therapy per year. This project is based on an entirely new concept that addresses the fundamental major limitations associated with delivery of any potential radiation mitigator of GI syndrome, including (i) non-invasive administration, (ii) targeted delivery of the therapeutic, (iii) maintained bioavailability of the therapeutic at efficacious dose, (iv) trivially scalable to produce, and (v) no need for formulation. The goal of this project is to develop an FDA-approved mitigator for radiation-induced GI injury. We propose critical studies to characterize the maximum effective dose, establish the rate of clearance from the body, and address safety by examining the effect of the drugs on the intestinal microbiome. In parallel, we will meet with the FDA to develop a strategy for IND approval that will guide subsequent studies. Successful completion of this work will place these probiotic drugs on a strong path towards eventual FDA approval.Project Narrative Mitigating normal tissue toxicity after exposure to ionizing radiation is critical to reduce mortality and morbidity, both for patients that receive pelvic or abdominal radiation therapy, and in the context of a RAD-NUC incident. We have developed second-generation probiotics engineered to express the therapeutic cytokines IL-22 or INF-β that have been shown to increase survival from 0–10% to 70–80%. The goal of this project is to continue to optimize and characterize these probiotic drugs for pivotal IND-enabling pre-clinical studies, which, if successful, will lead to the only FDA-approved radiation mitigators for GI toxicity.