Trudeau Institute Inc

CELL-MEDIATED PROTECTION AGAINST PNEUMONIC PLAGUE

REGULATION OF MEMORY CD8+ T CELL RECRUITMENT TO THE LUNG

T CELL MEMORY TO TB IN THE LUNG

T CELL RESPONSES TO CHRONIC BACTERIAL INFECTION

THE INTERDEPENDENCE BETWEEN T AND B CELLS IN TH2 CELL DIFFERENTIATION

ROLES FOR COAGULATION IN IMMUNITY

THE ROLE OF IL-4 AND IL-4RALPHA IN THE MULTI-STEP PROCESS OF TH2 DEVELOPMENT

THE ROLE OF IL-12P40 IN THE RESPONSE OF PULMONARY DENDRITIC CELLS TO TUBERCULOSIS

MEMORY T CELL SUBSETS AND PULMONARY IMMUNITY

ZIKA VIRUS COUNTERMEASURES PRE-CLINICAL PREGNANCY MODELS TO ASSESS PROTECTIVE EFFICACY AGAINST PLACENTAL DAMAGE AND FETAL DEMISE

DENDRITIC CELL FUNCTION IN SCHISTOSOMIASIS

AGING, T CELL REPERTOIRE, AND CELLULAR IMMUNITY TO INFLUENZA VIRUS

RESEARCH TRAINING IN IMMUNOLOGY AND INFECTIOUS DISEASES

IMMUNOGENICITY AND EFFICACY OF GENETICALLY ENGINEERED GAMMA-HERPESVIRUS VACCINES

OPTIMIZATION OF ANTITUBERCULAR COMPOUNDS AGAINST A NOVEL MYCOBACTERIUM TUBERCULOSIS TARGET, ASPC - TUBERCULOSIS (TB) IS ONCE AGAIN THE LEADING CAUSE OF DEATH FROM INFECTIOUS DISEASE. CURRENT TB TREATMENTS ARE COMPLEX, LENGTHY AND HAVE UNDESIRABLE SIDE EFFECTS. AS A RESULT, CASES OF MULTI-DRUG RESISTANT AND EXTENSIVELY-DRUG RESISTANT TB ARE ON THE RISE. COMPOUNDING THIS PROBLEM OF RISING DRUG RESISTANCE IS THE NOTABLE LACK OF INNOVATION IN DRUG DISCOVERY; NEW ANTIBIOTICS ARE OFTEN CHEMICALLY SIMILAR, TARGET THE SAME FEW BACTERIAL PATHWAYS (E.G. CELL WALL, NUCLEIC ACID AND PROTEIN SYNTHESIS) FOR WHICH MECHANISMS OF RESISTANCE ARE WELL ESTABLISHED, AND ARE SLOW TO COME TO MARKET. THEREFORE, THE DISCOVERY OF CHEMICALLY DISTINCT THERAPEUTICS THAT ACT VIA NOVEL MECHANISMS IS NECESSARY TO OVERCOME THE EMERGENCE OF RESISTANT STRAINS. IN ADDITION, DRUG REPURPOSING REPRESENTS AN ATTRACTIVE OPTION TO MITIGATE COST, TIME AND SAFETY RISK ASSOCIATED WITH ANTIBIOTIC DEVELOPMENT. WE HAVE IDENTIFIED A MOLECULE (TI-374) WITH HIGH POTENCY (0.1 ΜG/ML) AGAINST MYCOBACTERIUM TUBERCULOSIS (MTB), ORIGINALLY IDENTIFIED TO TREAT COGNITIVE IMPAIRMENT ASSOCIATED WITH SCHIZOPHRENIA AND PSYCHIATRIC DISORDERS. PRELIMINARY DATA INDICATES THAT THIS MOLECULE ACTS ON A NOVEL MTB TARGET, THE ALANINE AMINOTRANSFERASE ASPC, WHICH WAS OBSERVED TO BE ESSENTIAL IN A TRANSPOSON MUTAGENESIS SCREEN AND WE SHOW TO BE IMPORTANT FOR BACTERIAL SURVIVAL IN A MURINE MODEL OF MTB INFECTION. TI-374 HAS DRUG-LIKE AND LEAD- LIKE PROPERTIES, A NOVEL MECHANISM OF ACTION AND A DISTINCTIVE CHEMICAL STRUCTURE, THEREBY ADDRESSING THE MAJOR HURDLES PLAGUING ANTIBIOTIC DRUG DEVELOPMENT. WE HAVE PROPOSED TO OPTIMIZE TI-374 AGAINST MTB AND PROGRESS IT INTO A LEAD COMPOUND. KEY IDEAS UNDERLYING OUR APPROACH ARE AS FOLLOWS: (I) TAKING A LIGAND-BASED APPROACH, WE WILL EXPAND STRUCTURE-ACTIVITY RELATIONSHIP (SAR) AND OPTIMIZE POTENCY FOR EFFICACY AGAINST MTB ASPC USING ANTIBIOTIC SUSCEPTIBILITY ASSAYS AS WELL AS ON-TARGET ASPC ENZYMATIC ASSAYS AND OFF-TARGET ENZYMATIC ASSAYS TO COUNTER SCREEN AGAINST THE ORIGINAL HUMAN TARGET AND A KNOWN HUMAN ORTHOLOG, (II) ADVANCEMENT OF POTENT, ON- TARGET ANALOGS TO EXTENSIVE IN VITRO ADME AND IN VIVO PK STUDIES AND (III) CHARACTERIZATION OF PHARMACOLOGICAL AND GENETIC INHIBITION OF ASPC IN A MURINE CHRONIC INFECTION MODEL. WITH COMPLETION OF THIS PROPOSAL, WE EXPECT TO IDENTIFY 1-2 ADVANCED LEADS, AS WELL AS VALIDATE ASPC AS A NOVEL DRUGGABLE TARGET, ENABLING FURTHER STUDIES FOR THE TREATMENT OF MTB. WE BELIEVE THE PROPOSAL PROVIDES AN INNOVATIVE APPROACH TO DEVELOP NEW THERAPEUTIC AVENUES TO TREAT MTB INFECTIONS, WHICH IS AN URGENT UNMET NEED.

AN IMPROVED MOUSE MODEL FOR AGING IMMUNOLOGY

ANALYSIS OF CD8 T CELL RESPONSES TO TUMORS

DEVELOPMENT OF A MOUSE MODEL OF TUBERCULOSIS THAT GENERATES HUMAN-LIKE PATHOLOGY - ABSTRACT THOUGH VERY FEW NEW TUBERCULOSIS (TB) DRUGS HAVE BEEN INTRODUCED OVER THE PAST FORTY YEARS, THE PIVOTAL QUESTIONS RELEVANT TO TB THERAPEUTIC DEVELOPMENT ARE WELL ARTICULATED ACROSS THE FIELD AND ARE BEING PURSUED WITH GREAT VIGOR IN THE BIOMEDICAL COMMUNITY. CAN WE FIND A WAY TO SHORTEN TREATMENT FOR FROM THE CURRENT SIX MONTHS? WHICH NOVEL MECHANISMS OF ACTION WILL SUPPORT COMPLETE STERILIZATION WHEN COMBINED WITH EXISTING STANDARD OF CARE TREATMENT? ARE PERSISTER POPULATIONS COMPLETELY ELIMINATED THROUGH A SPECIFIC TREATMENT REGIMEN? IN ORDER TO SUCCEED IN THE CLINIC, FIRST WE MUST ADDRESS THESE QUESTIONS PRECLINICALLY THROUGH THE USE OF PREDICTIVE ANIMAL MODELS. EVALUATION OF THE EFFICACY OF EXPERIMENTAL TUBERCULOSIS THERAPEUTICS IS COMMONLY PERFORMED USING MOUSE MODELS, DUE TO THE RELATIVELY SMALL SIZE AND LOW COST OF MICE. HOWEVER, MANY MOUSE MODELS FAIL TO REPRODUCIBLY DEVELOP HUMAN-LIKE PATHOLOGY, SPECIFICALLY HYPOXIC NECROTIC GRANULOMAS, AND THEREBY MAY HAVE LIMITED PREDICTIVE VALUE FOR CERTAIN COMPOUNDS. MOUSE MODELS WHICH DO DEVELOP HUMAN-LIKE PATHOLOGY MAY SUFFER FROM SUBSTANTIAL HETEROGENEITY OR COMPLEX PROTOCOLS REQUIRED TO LIMIT SIGNIFICANT MORBIDITY. OUR RECENT WORK HAS OVERCOME THESE SIGNIFICANT LIMITATIONS, AND HERE WE PROPOSE TO REFINE, VALIDATE AND CHARACTERIZE AN INNOVATIVE MOUSE MODEL OF TB THAT IS MORE PREDICTIVE OF OUTCOMES IN PATIENTS. WE HAVE COMPLETED A SERIES OF STUDIES IN MICE THAT STRENGTHEN OUR ABILITY TO ADVANCE A MORE PREDICTIVE MODEL OF TB, ONE THAT CONSISTENTLY PRODUCES HUMAN-LIKE PATHOLOGY IN THE FORM OF HYPOXIC NECROTIC GRANULOMAS. THIS NEW MODEL BUILDS ON WORK THAT DEMONSTRATED THE DEVELOPMENT OF HYPOXIC GRANULOMAS IN NOS2-/-MICE, BUT ADDS SIGNIFICANT VALUE BY SIMPLIFYING THE PROTOCOL AND UTILIZING THE NATURAL AEROSOL ROUTE TO INFECT MICE WITH THE PARTIALLY ATTENUATED MYCOBACTERIUM TUBERCULOSIS (MTB) STRAIN R1RV. WE HAVE PAIRED THIS NOVEL MODEL WITH FLUORESCENT REPORTER STRAINS THAT SPECIFICALLY MARK DRUG AND IMMUNE TOLERANT MTB CELLS. WHEN DEPLOYED TOGETHER TO SUPPORT PHARMACOLOGICAL EVALUATION OF POTENTIAL TB DRUG REGIMENS, THESE TWO TOOLS CAN PROVIDE UNIQUE INSIGHTS INTO WHY SOME THERAPEUTIC APPROACHES MAY WORK WELL TO SHORTEN CHEMOTHERAPY IN PATIENTS, WHILE OTHERS WILL NOT DO SO. IN THIS PROPOSAL, WE SEEK TO ESTABLISH THE PREDICTIVE VALUE OF THE MODEL TO DETERMINE DRUG EFFICACY RESPONSES, AND TO REVEAL NEW INSIGHTS INTO THE BIOLOGY OF DRUG-TOLERANT MTB. IN THE FIRST TWO SPECIFIC AIMS WE SEEK TO OPTIMIZE THE NOVEL MOUSE MODEL AND VALIDATE IT AS A MORE RIGOROUS MEASURE OF IN VIVO ANTI-TUBERCULAR DRUG EFFICACY, AND IN THE THIRD AIM WE EMPLOY THE MODEL TO IDENTIFY THE NICHE OCCUPIED BY DRUG TOLERANT CELLS, TO BETTER UNDERSTAND THEIR POTENTIAL VULNERABILITIES. COMPLETION OF THE THREE AIMS OF THIS PROPOSAL WILL DELIVER A MORE PREDICTIVE AND WELL-CHARACTERIZED MOUSE MODEL OF MTB INFECTION THAT WILL HAVE THE POTENTIAL TO BECOME A NEW STANDARD FOR SMALL ANIMAL TESTING OF TB DRUGS.

A NOVEL DRUG RETARGETING PLATFORM FOR DRUG-RESISTANT BACTERIAL INFECTIONS

ANTIGEN-INDEPENDENT ACTIVATION OF MEMORY CD8 T CELLS DURING RESPIRATORY INFECTION

CAN PERSISTENT GAMMA-HERPESVIRUSES BE PURGED FROM THE HOST?

DEPOTS OF PERSISTENT INFLUENZA ANTIGENS SUPPORT PROTECTIVE CD4 T CELL MEMORY

MOFLO XDP HIGH SPEED CELL SORTER

CELL-MEDIATED PROTECTION AGAINST PNEUMONIC PLAGUE

CD8 MEMORY T CELLS: MECHANISMS OF PROTECTION

IMPACT OF AGING ON THE T CELL REPERTOIRE AND CELLULAR IMMUNITY TO INFLUENZA VIRUS

PROBING THE NOVEL MECHANISM OF ACTION OF TUBERCULOCIDAL COMPOUNDS - ABSTRACT TUBERCULOSIS (TB) IS ONCE AGAIN THE LEADING CAUSE OF DEATH FROM INFECTIOUS DISEASE. ENDING THE TB PANDEMIC REQUIRES NEW DRUGS THAT CAN STERILIZE RECALCITRANT PERSISTER CELLS. CURRENT TB TREATMENTS ARE COMPLEX, LENGTHY AND HAVE UNDESIRABLE SIDE EFFECTS. AS A RESULT, CASES OF MULTI-DRUG RESISTANT AND EXTENSIVELY-DRUG RESISTANT TB ARE ON THE RISE. COMPOUNDING THIS PROBLEM OF RISING DRUG RESISTANCE IS THE NOTABLE LACK OF INNOVATION IN DRUG DISCOVERY; NEW ANTIBIOTICS ARE OFTEN CHEMICALLY SIMILAR, TARGET THE SAME FEW BACTERIAL PATHWAYS (E.G. CELL WALL, NUCLEIC ACID AND PROTEIN SYNTHESIS) FOR WHICH MECHANISMS OF RESISTANCE ARE WELL ESTABLISHED, AND ARE SLOW TO COME TO MARKET. THEREFORE, THE DISCOVERY OF CHEMICALLY DISTINCT THERAPEUTICS THAT ACT VIA NOVEL MECHANISMS IS NECESSARY TO OVERCOME THE EMERGENCE OF RESISTANT STRAINS. FURTHER, DRUG REPURPOSING REPRESENTS AN ATTRACTIVE OPTION TO MITIGATE COST, TIME AND SAFETY RISK ASSOCIATED WITH ANTIBIOTIC DEVELOPMENT. WE HAVE IDENTIFIED A MOLECULE (TI-507), INITIALLY DISCOVERED AS A HUMAN SPHINGOSINE KINASE INHIBITOR, THAT IS ACTIVE AGAINST MYCOBACTERIUM TUBERCULOSIS (MTB) WITH AN IC50 OF 22 ΜM, KILLS PERSISTER CELLS AND STERILIZES CULTURES WITHIN 4 WEEKS. PRELIMINARY DATA SUGGESTS THAT THIS MOLECULE WORKS THROUGH A NOVEL MTB TARGET, NAD KINASE (PPNK), WHICH IS ESSENTIAL AND INVOLVED IN MANY CRITICAL CELLULAR PROCESSES. FURTHER, TI-507 HAS A DISTINCT CHEMICAL STRUCTURE AND DRUG-LIKE PROPERTIES, THEREBY ADDRESSING THE MAJOR HURDLES PLAGUING ANTIBIOTIC DRUG DEVELOPMENT. WHILE PPNK IS THOUGHT TO BE A PROMISING THERAPEUTIC TARGET, LITTLE IS KNOWN REGARDING THE BIOLOGICAL CONSEQUENCES OF PPNK INHIBITION IN MTB. AS SUCH, THIS PROPOSAL HAS BEEN DIVIDED INTO TWO SPECIFIC AIMS. AIM 1 IS FOCUSED ON TARGET VALIDATION AND UNDERSTANDING THE IMPORTANCE OF PPNK INHIBITION IN THE CONTEXT OF ENVIRONMENTAL STRESS, AN INTRACELLULAR INFECTION MODEL AND A MURINE INFECTION MODEL. AIM 2 FOCUSES ON EARLY STRUCTURE-ACTIVITY WORK INTENDED TO PROBE AND IMPROVE THE MOLECULE'S ANTI-TUBERCULAR ACTIVITY. WE BELIEVE THE PROPOSAL IS PROVIDING AN INNOVATIVE APPROACH TO DEVELOP NEW THERAPEUTIC AVENUES TO TREAT MTB INFECTIONS, WHICH IS AN URGENT UNMET NEED.

INKT AND B CELL COOPERATION IN IMMUNITY AND HOST DEFENSE

FUNCTIONAL CLONING OF S. MANSONI EGG-SPECIFIC TH2 CELLS

INHALED DNASE AS AN ADJUNCTIVE TUBERCULOSIS THERAPY

PROTECTIVE ROLES OF T CELLS DURING CAMPYLOBACTER COLONIZATION AND INFECTION

ROLE OF TGF-BETA IN SCHISTOSOMA MANSONI

CD* T CELL RESPONSES TO GAMMA-HERPESVIRAL INFECTION

IL12RBETA1 ALTERNATIVE SPLICE VARIANT IN THE REGULATION OF IL12 FAMILY SIGNALING

THE FUNCTION OF SAP IN T CELL HELP FOR B CELLS

MUCOSAL IMMUNITY: A TRUDEAU INSTITUTE WORKSHOP

THE YIN AND YANG OF INFLAMMATION