The goal of the proposed project is to develop a probiotic for the treatment of constipation, comprised of 2-3 anti-inflammatory strains of human gut-derived bacteria that enhance host serotonin (5-hydroxytryptamine, 5-HT)production and signaling. By addressing multiple targets associated with the etiology and symptoms ofconstipation (5-HT signaling and inflammation), we expect the result of this project to have broad utility.Constipation affects up to 20% of the general population, with rates even higher in the elderly, and comes inmany forms ranging in severity. The most severe form, slow transit constipation (STC), affects ~15 to 30% ofconstipated patients. Less severe, but still representing a significant societal burden, are irritable bowelsyndrome with constipation (IBS-C) and age-associated constipation. Constipation is also a major side effect ofopioid medications, and is comorbid with neurological diseases including Parkinsonian syndromes. The etiologyis multifarious, often involving a dysfunctional enteric nervous system, poor diet, and inflammation, but in manycases it is still poorly understood. Current treatment options are generally poor and include dietary modifications,pharmacological interventions, and-for severe forms like STC-surgery. As such, novel treatment options areneeded, preferably those that can engage multiple therapeutic targets. One source of new therapeutics is thegut microbiome: the bacteria that reside in the gastrointestinal tract. These symbiotic organisms are involved innumerous components of health and disease, including development and maintenance of the immune andenteric nervous systems. Recently, it has been shown by a member of Holobiome's team that the gut microbiomeis a major regulator of host enteric 5-HT signaling: Germ free mice have a 50% reduction in serum 5-HT andhave constipation, both of which can be corrected via recolonization with a normal microbiome or a human-derived consortium. It has also been shown that constipation in humans can be transferred into animals via fecalmicrobiome transplant; a phenotype that is driven via disrupted enteric 5-HT signaling and inflammation. Thissuggests a clear intervention strategy: deliver anti-inflammatory, 5-HT-modulating bacteria to the gut. In ourpreliminary studies we screened a proprietary panel of over 100 non-pathogenic species of human gut bacteriafor the ability to modulate 5-HT in a cell culture model. We identified 17 bacterial strains with the capability tomodulate 5-HT signaling via four mechanisms. In this Phase 1 application we seek to further test these strainsas candidates for a 5-HT modulating and anti-inflammatory consortium. To do this, strains will first be profiled forsafety and manufacturability in vitro and in silico. Strains meeting these criteria will be assembled into candidateconsortia of 2-3 strains, which will be further optimized to impact 5-HT release and inflammation. These will beadvanced for testing in mouse models of constipation to assess efficacy, safety, and engraftment. The mostpromising consortium will be further tested in PK/PD and dosing studies supported in a Phase 2 program.
Public Health Relevance Statement: The goal of this project is to develop new therapies for constipation. Bacteria, isolated from healthy humans, will
be tested for the ability to positively impact potential causes of constipation and its associated symptoms in cell
culture and in vivo models, which are predictive of effectiveness in humans.
Project Terms: Affect ; Age ; ages ; Elderly ; advanced age ; elders ; geriatric ; late life ; later life ; older adult ; older person ; senior citizen ; Animals ; Anti-Inflammatory Agents ; Anti-Inflammatories ; Anti-inflammatory ; Antiinflammatories ; Antiinflammatory Agents ; antiinflammatory ; Antibiotics ; Antibiotic Agents ; Antibiotic Drugs ; Miscellaneous Antibiotic ; Bacteria ; Biological Assay ; Assay ; Bioassay ; Biologic Assays ; Body Weight ; Cecum ; Cell Culture Techniques ; cell culture ; cell motility ; Cell Locomotion ; Cell Migration ; Cell Movement ; Cellular Migration ; Cellular Motility ; Motility ; Cells ; Cell Body ; comorbidity ; co-morbid ; co-morbidity ; Constipation ; Diet ; diets ; Disease ; Disorder ; Dyes ; Coloring Agents ; Eating ; Food Intake ; Exhibits ; Feces ; stool ; Freeze Drying ; Freeze Dryings ; Lyophilization ; Freezing ; Future ; Gastrointestinal tract structure ; Alimentary Canal ; Digestive Tract ; GI Tract ; Gastrointestinal Tract ; alimentary tract ; digestive canal ; Gene Expression ; Goals ; Growth ; Generalized Growth ; Tissue Growth ; ontogeny ; Health ; Human ; Modern Man ; In Vitro ; Inflammation ; Intestinal Mucosa ; Intestines ; Intestinal ; bowel ; Inulin ; Irritable Bowel Syndrome ; Irritable Colon ; Mucous Colitis ; spastic colon ; Maintenance ; Morphine ; Infumorph ; Kadian ; MS Contin ; MSir ; Morphia ; Oramorph ; Oramorph SR ; Roxanol ; Statex SR ; Mus ; Mice ; Mice Mammals ; Murine ; nervous system disorder ; Nervous System Diseases ; Neurologic Disorders ; Neurological Disorders ; neurological disease ; Neurobiology ; neurobiological ; Organism ; living system ; Organoids ; Patients ; Pharmacology ; Phenotype ; Production ; Safety ; Serotonin ; 5-HT ; 5-Hydroxytryptamine ; 5HT ; Enteramine ; Hippophaine ; Signal Transduction ; Cell Communication and Signaling ; Cell Signaling ; Intracellular Communication and Signaling ; Signal Transduction Systems ; Signaling ; biological signal transduction ; Technology ; Testing ; Tissues ; Body Tissues ; Toxin ; Tryptamines ; Indolylethylamines ; cytokine ; Measures ; Diet Modification ; Dietary Modifications ; diet alteration ; dietary alteration ; Serotonin Agonists ; 5-HT Agonists ; 5-Hydroxytryptamine Agonists ; 5-Hydroxytrytamine Agonist ; Serotonergic Agonists ; Serotonin Receptor Agonists ; base ; Blood specimen ; Blood Sample ; Clinical ; Phase ; Enteric Nervous System ; Predisposition ; Susceptibility ; Fiber ; Serum ; Blood Serum ; Individual ; Recovery ; Opioid ; Opiates ; Parkinsonian Disorders ; Parkinsonian ; Parkinsonian Condition ; Parkinsonian Diseases ; Parkinsonian Syndrome ; Parkinsonism ; Agonist ; 5-HT4 Receptor ; 5-Hydroxytryptamine-4 Receptor ; Serotonin 4 Receptor ; Serotonin Receptors 5-HT4 ; Engraftment ; Therapeutic ; fluid ; liquid ; Liquid substance ; L rhamnosus ; L. rhamnosus ; Lactobacillus rhamnosus ; Lactobacillus casei rhamnosus ; Exposure to ; Morphology ; Inflammatory ; programs ; Immunes ; Immune ; Severities ; Oral ; subdermal ; subcutaneous ; microorganism ; Source ; Probiotics ; Operative Procedures ; Surgical ; Surgical Interventions ; Surgical Procedure ; surgery ; Operative Surgical Procedures ; meetings ; experience ; synergism ; PBMC ; Peripheral Blood Mononuclear Cell ; novel ; member ; General Public ; General Population ; Modality ; Modeling ; LC/MS ; liquid chromatography mass spectrometry ; Intervention Strategies ; interventional strategy ; Intervention ; Effectiveness ; Enteric ; Enteral ; Causality ; causation ; disease causation ; Etiology ; genome sequencing ; Address ; food monitoring ; food supply monitoring ; food supply surveillance ; food surveillance ; Dose ; Symptoms ; in vivo Model ; Germ-Free ; Immunologics ; Immunochemical Immunologic ; Immunologic ; Immunological ; Immunologically ; Development ; developmental ; Output ; microbiome ; feeding ; prebiotics ; Antibiotic susceptibility ; Population ; metagenomic sequencing ; metagenome sequencing ; novel therapeutics ; new drug treatments ; new drugs ; new therapeutics ; new therapy ; next generation therapeutics ; novel drug treatments ; novel drugs ; novel therapy ; mouse model ; murine model ; stem ; therapeutic target ; effective therapy ; effective treatment ; prescription opioid ; licit opioid ; opiate medication ; opioid medication ; prescribed opiate ; prescribed opioid ; prescription opiate ; arm ; screening ; gut microbiome ; GI microbiome ; digestive tract microbiome ; enteric microbiome ; gastrointestinal microbiome ; gut-associated microbiome ; intestinal biome ; intestinal microbiome ; fecal transplantation ; fecal microbial transplantation ; fecal microbiome transplantation ; fecal microbiota transplant ; fecal microbiota transplantation ; fecal transplant ; associated symptom ; co-morbid symptom ; co-occuring symptom ; comorbid symptom ; concurrent symptom ; cooccuring symptom ; symptom association ; symptom comorbidity ; therapeutic candidate ; manufacturability ; treatment arm ; intervention arm ; gut bacteria ; bacteria in the gut ; Unhealthy Diet ; poor diet ; microbiome sequencing ; side effect ; pharmacokinetics and pharmacodynamics ; PK/PD ; in silico ;
