The world is rapidly heading towards a pre-1940's scenario when it comes to fighting infectious disease. Antimicrobial resistance is a growing problem on a global scale, greatly hampering our abilities to quell worldwide epidemics such as tuberculosis and malaria, as well as the simple staphylococcus infection . The proposed project is significant because unless innovative strategies are developed to produce robust and effective new classes of antibiotics, health care costs will continue to climb and we will completely lose our ability to combat even the most common infection. Current antibiotic treatments originated predominantly from natural products produced by fungi and bacteria that were able to inhibit the growth of other organisms, usually by inhibiting cell wall synthesis or maintenance or by inhibiting protein synthesis. Since penicillin was first isolated by Fleming in 1929, most of the subsequent generations of antibiotics remain very similar to the original natural products, wit h functional groups modified to increase their activity across a broader range of pathogens and decrease their side effect profiles. Oxazolidones, glycopeptides, ï¢-lactams, and quinolones show some promise for the future, but Gram-negative bacterial infections still remain problematic. Cases of multidrug-resistant (MDR, resistance to 2-3 classes), extensive drug resistance (XDR, resistance to most classes except colistin or tigecycline) and even pan drug resistance (PDR, resistance to all classes) nosocomial bacterial infections have skyrocketed in recent years, and the emergence of pan drug-resistant isolates are making these infections increasingly difficult to treat. Hospital-acquired infections like these account for up to 4% of all hospital stays in the United States and are incredibly diverse in causative pathogen, antibiotic resistance profile, and severity. A significant cause of nosocomial infection is the Enterobacteriaceae family, which includes Gram-negative bacilli that can be commensal or pathogenic. Enterobacteriaceae have a widespread clinical and economic impact due to the diversity of infections they cause; this family causes many infections such as pneumonia, bloodstream infections (BSIs), urinary tract infections (UTIs), and intra- abdominal infections (IAIs). The World Health Organization (WHO) lists carbapenem-resistant Enterobacteriaceae (CRE) as having a critical need for novel antibiotics on their Priority Pathogens list. Because the mortality of these multi drug-resistant infections is between 30 and 50% and there is such difficulty in finding viable treatments, the need for novel therapeutics for these pathogens must be addressed. Nucleic acids are promising avenues for drug design, both as therapeutics and as targets. Targeting heavily conserved RNA sequences and structures, in bacteria (Enterobacteriaceae), and involved in proliferation and survival of bacteria, is a promising approach. Using our proprietary probes, assays and libraries, we propose to develop a screening assay for an essential gene in Enterobacteriaceae. Here we propose an innovative plan for identification of a novel class of ligands that are specific for an mRNA present in an essential gene in bacteria, and we propose a biophysical screening assay for identifying such ligands. First, as outlined in Specific Aim 1, we will characterize a model nucleic acid domain that will be synthesized commercially and identify specific and high-affinity aminosugar binders. We will then synthesize sequence-specific RNA binding ligands and screen this targeted library of conjugates for sequence-specific binding and gene inhibition. The mechanism of action will be confirmed using a reporter gene assay (Specific Aim 2). A successful application of the approach will allow us to identify and validate lead compounds for inhibition of bacterial growth in Phase II studies.
Public Health Relevance Statement: PROJECT NARRATIVE Antimicrobial resistance occurs when microorganisms (often infectious bacteria, viruses, and certain parasites) are no longer sensitive to drugs that were previously used to treat them; this is of global concern because it hampers our ability to control infectious disease and increases the costs of health care. In order to combat this world-wide problem, innovative strategies for antibiotic drug design must be implemented. This project addresses this emergent problem by targeting an essential mRNA sequence in gram negative bacteria, by using a novel in-house biophysical screening assay to sequence-specifically block bacterial growth.
Project Terms: Amino Sugars; aminosugar; Antibiotics; Antibiotic Agents; Antibiotic Drugs; Miscellaneous Antibiotic; Awareness; Bacillus; Bacteria; Bacterial Infections; bacteria infection; bacterial disease; Base Sequence; Nucleotide Sequence; nucleic acid sequence; Biological Assay; Assay; Bioassay; Biologic Assays; Biophysics; biophysical foundation; biophysical principles; biophysical sciences; Boston; Cell membrane; Cytoplasmic Membrane; Plasma Membrane; plasmalemma; Cell Wall; Cells; Cell Body; Chemistry; High Pressure Liquid Chromatography; HPLC; High Performance Liquid Chromatography; High Speed Liquid Chromatography; Colistin; Colimycin; Colisticin; Polymyxin E; Communicable Diseases; Infectious Disease Pathway; Infectious Diseases; Infectious Disorder; Cessation of life; Death; DNA; Deoxyribonucleic Acid; Topoisomerase II; DNA Topoisomerase II; DNA Type 2 Topoisomerase; Topo II; Type II DNA Topoisomerases; Drug Design; Drug resistance; drug resistant; resistance to Drug; resistant to Drug; Microbial Drug Resistance; Anti-microbial Drug Resistance; Anti-microbial Drug Resistant; Antimicrobial Drug Resistance; Antimicrobial Drug Resistant; microbial drug resistant; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Enterobacteriaceae; Coliform Bacilli; Enteric Bacteria; Enterobacteria; Enzymes; Enzyme Gene; Epidemic; Escherichia coli; E coli; E. coli; Family; Fluorescence; fungus; Future; Genes; Glycopeptides; Gram-Negative Bacteria; Growth; Generalized Growth; Tissue Growth; ontogeny; HIV; AIDS Virus; Acquired Immune Deficiency Syndrome Virus; Acquired Immunodeficiency Syndrome Virus; Human Immunodeficiency Viruses; LAV-HTLV-III; Lymphadenopathy-Associated Virus; Virus-HIV; Hybrids; Infection; Institute of Medicine (U.S.); Institute of Medicine; NAS/IOM; Klebsiella; Calymmatobacterium; Donovania; Lactams; Lead; Pb element; heavy metal Pb; heavy metal lead; Length of Stay; Number of Days in Hospital; hospital days; hospital length of stay; hospital stay; Libraries; Ligands; Maintenance; Malaria; Paludism; Plasmodium Infections; Morbidity - disease rate; Morbidity; mortality; Persons; National Academy of Sciences; United States National Academy of Sciences; Nucleic Acids; living system; Organism; Parasites; Patents; Legal patent; Penicillins; Peptides; Play; Pneumonia; Ketoquinolines; Oxoquinolines; Quinolinones; Quinolones; DNA Recombination; Recombination; Genetic Recombination; Ribosomes; Non-Polyadenylated RNA; RNA Gene Products; Ribonucleic Acid; RNA; mRNA; Messenger RNA; Salmonella; Shigella; Staphylococcus infection; Staphylococcal Infections; Genetic Transcription; Gene Transcription; RNA Expression; Transcription; Translations; Tuberculosis; M tuberculosis infection; M. tb infection; M. tuberculosis infection; M.tb infection; M.tuberculosis infection; MTB infection; Mycobacterium tuberculosis (MTB) infection; Mycobacterium tuberculosis infection; TB infection; disseminated TB; disseminated tuberculosis; infection due to Mycobacterium tuberculosis; tuberculosis infection; tuberculous spondyloarthropathy; United States; Urinary tract infection; Urinary tract infectious disease; urinary infection; Virus; Wit; Work; World Health Organization; Generations; Antisense Oligonucleotides; Anti-Sense Oligonucleotides; Antisense Agent; anti-sense agent; anti-sense oligo; antisense oligo; Gram-Negative Bacterial Infections; Health Care Costs; Health Costs; Healthcare Costs; Antibiotic Resistance; Resistance to antibiotics; Resistant to antibiotics; antibiotic drug resistance; antibiotic resistant; RNA Sequences; base; improved; Solid; Clinical; Phase; Nosocomial Infections; Hospital Infections; Hospital acquired infection; institutional infection; Reporter Genes; Gene Targeting; Multidrug Resistance; Multiple Drug Resistance; Multiple Drug Resistant; Resistance to Multi-drug; Resistance to Multidrug; Resistance to Multiple Drug; Resistant to Multiple Drug; Resistant to multi-drug; Resistant to multidrug; multi-drug resistant; multidrug resistant; Multi-Drug Resistance; analog; uptake; Antibacterial Agents; anti-bacterial; antibacterial; Anti-Bacterial Agents; Therapeutic; Reporter; Antibiotic Treatment; bacterial disease treatment; bacterial infectious disease treatment; Antibiotic Therapy; fighting; anti-microbial agent; anti-microbial drug; antimicrobial agent; antimicrobial drug; Severities; microorganism; System; interest; Ribosomal Peptide Biosynthesis; Ribosomal Protein Biosynthesis; Ribosomal Protein Synthesis; protein synthesis; Protein Biosynthesis; functional group; DNA Replication; DNA Synthesis; DNA biosynthesis; Essential Genes; Peptide Nucleic Acids; Toxicities; Toxic effect; Structure; novel; member; economic impact; Coding System; Code; Modeling; immunogenic; Bacterial Antibiotic Resistance; antibiotic resistant bacteria; bacterial antibiotic resistant; bacterial resistance to antibiotic; DNA Gyrase; MicroRNAs; Micro RNA; miRNA; miRNAs; Abdominal Infection; Post-Transcriptional Gene Silencing; Posttranscriptional Gene Silencing; Quelling; RNA Silencing; RNAi; Sequence-Specific Posttranscriptional Gene Silencing; RNA Interference; Pathogenicity; Nucleic Acid Binding; RNA bound; RNA Binding; Molecular Interaction; Binding; GAR-936; tigecycline; RNA synthesis; RNA chemical synthesis; Address; Length; Antimicrobial resistant; Resistance to antimicrobial; anti-microbial resistance; anti-microbial resistant; resistance to anti-microbial; resistant to anti-microbial; resistant to antimicrobial; Antimicrobial Resistance; Affinity; Data; Intra-abdominal; Ligand Binding; Development; developmental; cost; Sepsis; blood infection; bloodstream infection; novel strategies; new approaches; novel approaches; novel strategy; targeted delivery; site targeted delivery; Resistance profile; Resistant profile; pathogen; innovation; innovate; innovative; Resistance; resistant; antimicrobial; anti-microbial; novel therapeutics; new drug treatments; new drugs; new therapeutics; new therapy; next generation therapeutics; novel drug treatments; novel drugs; novel therapy; combat; alternative treatment; phase 2 study; phase II study; screening; carbapenem-resistant Enterobacteriaceae; extensive drug resistance; extensively drug resistant; extreme drug resistance; Natural Products; targeted agent; experimental study; experiment; experimental research; priority pathogen; side effect; antibiotic resistant infections; off-target site; Rapid screening; novel antibiotic class; new antibiotic class; new antibiotic type
