Sepsis results in massive loss of life and places a significant economic burden on society. There are no effective treatments available for human sepsis other than antibiotics and life support. It is increasingly clear that sepsis is a bi-phasic process comprised of 1) an early high-energy demanding hyperinflammation state that can cause inflammatory shock and 2) a low energy supply immunosuppression state that promotes immunometabolic paralysis while countering oxidative damage. These two phases are seamlessly connected or even concurrent. This makes sepsis treatment extremely difficult, and many therapies such as anti- inflammatory corticosteroids often worsen the outcome. Macrophages (M?s) play essential roles throughout the course of sepsis. In the hyperinflammation phase, M?s sense pathogen-associated molecular patterns (PAMPs) through receptors such as toll-like receptors (TLRs) and NOD-, LRP-, and pyrin domain-containing protein 3 (NLRP3). M? NLRP3 inflammasome activation and resulting IL-1? secretion cause acute organ damage and release of damage-associate molecular patterns (DAMPs), which act back on the inflammatory pathways, forming a vicious cycle. Therefore, the M? NLRP3 inflammasome is a major contributor to the hyperinflammation phase of bacterial sepsis. Concomitant with inflammasome activation, M?s undergo a broad cellular metabolic rewiring that favors glycolysis and turns mitochondria from ATP generation to reactive oxygen species (ROS) production, leading to mitochondrial oxidative stress, metabolic paralysis, and M? anergy in the immunosuppression phase. In addition, NLRP3 inflammasome activation results in GSDMD- mediated pyroptotic cell death (pyroptosis), directly removing M?s from the fight against secondary infections. Recently, we identified that in M?s, pyruvate dehydrogenase kinase 1 (PDHK1) plays a critical role in coordinating inflammasome activation and metabolic rewiring. In M?s treated with LPS and ATP or Nigericin to stimulate inflammasome activation, dichloroacetate (DCA, a pyruvate analog and pan-PDHK inhibitor) or JX06 (a synthetic small-molecule and selective PDHK1 inhibitor) effectively suppressed IL-1? secretion and cell death, improved mitochondrial integrity, and reprogramed mitochondria from ROS production to ATP generation. In a mouse cecal ligation and puncture (CLP) model, PDHK inhibition significantly reduced plasma IL-1? levels. In this STTR Phase 1 project, we will test the hypothesis that JX06 can be developed as a novel therapy for bacterial sepsis. We propose two specific aims: SA1. To determine the toxicity of JX06 in cultured primary mouse and human cells in vitro and mice in vivo, and to study its pharmacokinetics in mice. SA2. To establish the effectiveness of JX06 in various mouse strains using the CLP model of sepsis. This STTR Phase 1 project will validate the role of M? PDHK1 in bacterial sepsis and provide a proof of concept to develop JX06 or its analog as a new therapeutic agent for bacterial sepsis. The successful completion of these proposed studies will serve as a milestone for the further development effort. Public Health Relevance Statement Narrative. Sepsis results in massive loss of life and places a significant economic burden on society. There are no effective treatments available for bacterial sepsis. We propose to develop JX06, a selective macrophage PDHK1 inhibitor, as a novel therapy for bacterial sepsis by suppressing macrophage NLRP3 inflammasome activation and restoring macrophage metabolic health.
Project Terms: Corticoids; Corticosteroids; Adrenal Cortex Hormones; inhibitor; Anti-Inflammatories; Anti-inflammatory; Antiinflammatories; Antiinflammatory Agents; antiinflammatory; Anti-Inflammatory Agents; Antibiotic Agents; Antibiotic Drugs; Miscellaneous Antibiotic; Antibiotics; Attention; Back; Dorsum; Bacteremia; bacteraemia; bacterial sepsis; Biological Availability; Bioavailability; Physiologic Availability; Bone Marrow; Bone Marrow Reticuloendothelial System; Brain; Brain Nervous System; Encephalon; Cell Death; necrocytosis; Cells; Cell Body; Cultured Cells; Dendritic Cells; Veiled Cells; Dichloroacetate; Disease; Disorder; Investigational Drugs; Investigational New Drugs; Energy Supply; Female; Glycolysis; Cyclic GMP; Guanosine Cyclic Monophosphate; cGMP; Health; Heart; Heterogeneity; Human; Modern Man; Immune system; Immunocompetence; Immunologic Competence; Immunological Competence; Immunosuppression; Immunosuppression Effect; Immunosuppressive Effect; immune suppression; immune suppressive activity; immune suppressive function; immunosuppressive activity; immunosuppressive function; immunosuppressive response; In Vitro; Inbreeding; Inflammation; Interleukin-1 beta; Beta Proprotein Interleukin 1; IL-1 beta; IL-1 ß; IL-1-b; IL-1ß; IL1-Beta; IL1-ß; IL1B Protein; IL1F2; IL1ß; Interleukin 1beta; Interleukin-1ß; Preinterleukin 1 Beta; Kidney; Kidney Urinary System; renal; Liver; hepatic body system; hepatic organ system; Macrophage; MÏ; male; Mitochondria; mitochondrial; Mus; Mice; Mice Mammals; Murine; Nigericin; Oxidative Phosphorylation; Oxidative Phosphorylation Pathway; Phagocytes; Phagocytic Cell; amebocyte; Drug Kinetics; Pharmacokinetics; Plasma; Blood Plasma; Plasma Serum; Reticuloendothelial System, Serum, Plasma; Play; Production; Proteins; Role; social role; Safety; Shock; Circulatory Collapse; circulatory shock; shocks; Societies; Survival Rate; Family suidae; Pigs; Suidae; Swine; porcine; suid; T-Lymphocyte; T-Cells; thymus derived lymphocyte; Testing; Time; Generations; Mediating; pyruvate dehydrogenase kinase; PDH kinase; Active Oxygen; Oxygen Radicals; Pro-Oxidants; Reactive Oxygen Species; Organ; improved; Acute; Phase; Histologically; Histologic; Biochemical; anergy; Hepatic Cells; Hepatic Parenchymal Cell; Liver Cells; Hepatocyte; Measurement; Oxidative Stress; Toxicity Testing; Toxicity Tests; analog; Pyruvate; Therapeutic; Therapeutic Agents; Metabolic; Attenuated; attenuate; attenuates; Inflammatory; Life; fighting; Immune; Immunes; secondary infection; Pattern; Paralysed; Palsy; Plegia; paralysis; paralytic; receptor; Receptor Protein; Animal Model; Animal Models and Related Studies; model of animal; Toxic effect; Toxicities; marenostrin; MEFV gene product; pyrin; Toll-like receptors; TLR protein; Toll-Like Receptor Family Gene; organ regeneration; Modeling; Vascular Endothelial Cell; Effectiveness; small molecule; Dose; Data; Economic Burden; Mononuclear; Mouse Strains; Resolution; resolutions; in vivo; Invaded; Small Business Technology Transfer Research; STTR; Molecular; Process; Development; developmental; Pathway interactions; pathway; pre-clinical; preclinical; blood infection; bloodstream infection; Sepsis; determine efficacy; efficacy analysis; efficacy assessment; efficacy determination; efficacy examination; evaluate efficacy; examine efficacy; efficacy evaluation; Outcome; pathogen; Microbe; new drug treatments; new drugs; new pharmacological therapeutic; new therapeutics; new therapy; next generation therapeutics; novel drug treatments; novel drugs; novel pharmaco-therapeutic; novel pharmacological therapeutic; novel therapy; novel therapeutics; mitochondrial dysfunction; murine model; mouse model; effective treatment; effective therapy; oxidative injury; oxidative damage; safety testing; phase II study; phase 2 study; Regimen; individual patient; Formulation; Inflammasome; sepsis patients; septic patients; CLP model; CLP mouse model; Cecal ligation perforation; cecal ligation and perforation; cecal ligation and puncture; cecum ligation and puncture; cecum ligation puncture; cecal ligation puncture; metabolic fitness; manufacture
