Irinotecan-based chemotherapy is widely used for the treatment of various types of metastatic cancers, however, severe drug-induced injury or gastrointestinal (GI) toxicity, especially severe delayed-onset diarrhea (SDOD) induced by SN-38 (an active metabolite of irinotecan), limits its clinical application of the drug. Irinotecan-induced SDOD incidences result in increased healthcare cost due to hospitalization, and poor quality of life and therapeutic outcomes for the cancer patients. Currently, there is no effective treatment available for about ~15% of patients receiving irinotecan therapy, who suffer unmanageable SDOD symptoms, mainly due to their inability to detoxify the SN-38 in the colon. SN-38 mediated intestinal toxicity was found to be more severe in patients with uridine diphosphate glycosyltransferases 1A1 (UGT1A1) polymorphism, delineating the critical role of colonic UGT1A1 in the prevention of SN-38 induced SDOD. Various approaches to reduce SN-38 colonic exposure has been tried without much success, and SDOD remains unmanageable dose-limiting toxicity of irinotecan in adult and pediatric cancer patients. Preliminary research of our academic research collaborators at the University of North Texas and the University of Houston discovered glucosyltransferases from plant Medicago truncatula that could efficiently metabolize typical human UGT1A1 substrates, including SN-38, in an in vitro assay. This led to the novel and innovative concept of bioengineering commensal bacteria E.coli (EC) for overexpressing plant UGT71G1 to create a drug detoxifying bacteria (DDB) that can detoxify SN-38. In this STTR project, we propose to generate proof-of-concept preclinical evidence in support of developing a safe, efficacious, and traceable DDB as a novel live biotherapeutic product (LBP) to alleviate and/or prevent the SN- 38-mediated intestinal toxicity. Here, we will bioengineer a commensal E.coli strain, with good human gut colonization potential, to overexpress the most active variant of UGT71G1 (U71G1*n) tagged with a green fluorescent protein (GFP), which will be delivered in a protective capsule directly to the colon for the effective glycosylation of SN-38 to SN-38-glucose. To achieve this goal, the proposed specific aims are 1) to develop traceable and more active drug detoxifying bacteria EC_U71G1*n (active variant) to detoxify SN-38, 2) to develop a colon-optimized capsule delivery of traceable and highly active lyophilized DDBs, and 3) to evaluate the efficacy of 2 colon-delivered DDBs in irinotecan induced SDOD rat model. An active DDB engineered with pUGT biocatalyst (for the detoxification of SN-38) and GFP (for tracing the effective gut colonization in human feces for therapeutic efficacy monitoring) will accelerate the recovery of cancer patients from SDOD, and lead to the effective management of irinotecan-induced SDOD in clinics. The success of this project will provide Sanarentero with the proof-of-concept evidence needed for developing an SN-38-targeted traceable DDB for preclinical and IND-enabling studies in a phase-II SBIR/STTR application. The novel LBP will act as a complementary therapy to treat SDOD, which should improve the quality of life and even therapeutic outcomes for metastatic cancer patients. Once successful in this endeavor, we plan to apply the same approach to utilize different plant UGTs for inactivating other GI-toxic drugs, xenobiotics, pollutants, or their toxic metabolites in the colon by developing drug-specific DDB. Public Health Relevance Statement NarrativeThis project develops a novel live biotherapeutic product (LBP) for attenuating irinotecan chemotherapy- induced severe and delayed onset diarrhea. Our novel LBP uses an innovative concept of bioengineered drug detoxifying bacteria (DDB) expressing plant glucosyltransferases to detoxify SN-38 (a gut-toxic metabolite of irinotecan) in the colon. We will deliver our safe and efficacious DDB using a protective colon-optimized delivery system to maximize its efficacy and colonization.
Project Terms: Adult ; 21+ years old ; Adult Human ; adulthood ; Bacteria ; Biological Response Modifier Therapy ; Biologic Therapy ; Biological Therapy ; biological therapeutic ; biological treatment ; biotherapeutics ; biotherapy ; Biomedical Engineering ; bio-engineered ; bio-engineers ; bioengineering ; capsule ; Capsules ; Colon ; Diarrhea ; Pharmaceutical Preparations ; Drugs ; Medication ; Pharmaceutic Preparations ; drug/agent ; Engineering ; Enzymes ; Enzyme Gene ; Escherichia coli ; E coli ; E. coli ; Feces ; stool ; Fluorescence ; Freeze Drying ; Freeze Dryings ; Lyophilization ; Gastric Acid ; Gastric Hydrochloric Acid ; Glucose ; D-Glucose ; Dextrose ; Glucosyltransferases ; glycosylation ; Metabolic Glycosylation ; Goals ; Hospitalization ; Hospital Admission ; Human ; Modern Man ; Incidence ; Intestines ; Intestinal ; bowel ; Lead ; Pb element ; heavy metal Pb ; heavy metal lead ; Drug Metabolic Detoxication ; Drug Metabolic Detoxification ; Metabolic Drug Detoxications ; Metabolism of Toxic Agents ; detoxification ; Patients ; Plant Proteins ; Plants ; Polymers ; Genetic Polymorphism ; polymorphism ; Proteins ; Quality of life ; QOL ; Rattus ; Common Rat Strains ; Rat ; Rats Mammals ; Research ; Role ; social role ; Safety ; Technology ; Texas ; Universities ; Uridine Diphosphate ; Uridine Pyrophosphate ; Xenobiotics ; Fluorescent Protein Tracings ; Measures ; glycosyltransferase ; EC 2.4 ; Glycoside Transferases ; Health Care Costs ; Health Costs ; Healthcare Costs ; Mediating ; Treatment Cost ; Green Fluorescent Proteins ; irinotecan ; Campto ; camptosar ; SN-38 ; Injury ; injuries ; base ; improved ; Site ; Clinical ; Encapsulated ; Phase ; Variant ; Variation ; Medical ; Series ; Distal part of ileum ; Terminal Ileum ; Recovery ; Malignant Childhood Neoplasm ; Childhood Cancers ; Malignant Childhood Tumor ; Malignant Pediatric Neoplasm ; Malignant Pediatric Tumor ; Malignant childhood cancer ; cancer in a child ; cancer in children ; child with cancer ; childhood malignancy ; children with cancer ; pediatric cancer ; pediatric malignancy ; Collaborations ; Medicago truncatula ; Attenuated ; Metastatic Cancer ; Metastatic Malignant Neoplasm ; Disseminated Malignant Neoplasm ; Hour ; Clinic ; System ; gastrointestinal ; GNT1 ; UDP glycosyltransferase 1 family, polypeptide A1 Gene ; UDP glycosyltransferase gene 1 ; UGT1 ; UGT1A ; UGT1A1 ; UGT1A5 ; UGT1A1 gene ; success ; pollutant ; Toxicities ; Toxic effect ; novel ; Prevention ; Modeling ; drug development ; Complementary treatment ; Complementary therapies ; preventing ; prevent ; Dose ; Symptoms ; Dose-Limiting ; Recombinants ; in vitro Assay ; in vivo ; Cancer Patient ; therapy outcome ; therapeutic outcome ; Small Business Innovation Research Grant ; SBIR ; Small Business Innovation Research ; Small Business Technology Transfer Research ; STTR ; Monitor ; Process ; pre-clinical ; preclinical ; design ; designing ; efficacy evaluation ; efficacy analysis ; efficacy assessment ; efficacy examination ; evaluate efficacy ; examine efficacy ; Treatment Efficacy ; intervention efficacy ; therapeutic efficacy ; therapy efficacy ; innovation ; innovate ; innovative ; clinical application ; clinical applicability ; chemotherapy ; novel therapeutics ; new drug treatments ; new drugs ; new therapeutics ; new therapy ; next generation therapeutics ; novel drug treatments ; novel drugs ; novel therapy ; overexpression ; overexpress ; FDA approved ; effective therapy ; effective treatment ; preclinical efficacy ; pre-clinical efficacy ; gut microbiota ; GI microbiota ; Gastrointestinal microbiota ; enteric microbial community ; enteric microbiota ; gastrointestinal microbial flora ; gut commensal ; gut community ; gut flora ; gut microbe community ; gut microbial community ; gut microbial composition ; gut microbial consortia ; gut microbiotic ; gut microflora ; intestinal flora ; intestinal microbes ; intestinal microbiota ; intestinal microflora ; intestinal tract microflora ; screening ; commensal bacteria ; commensal bacterial species ; gut colonization ; GI colonization ; gastrointestinal tract colonization ; intestinal colonization ;
