University Of Western Ontario

IMPACT OF HIV-1 FITNESS ON DISEASE PROGRESSION

HYPERPOLARIZED 13C MRI OF PLACENTAL METABOLIC ABNORMALITIES RESULTING FROM THE WESTERN DIET

VIRAL AND HOST FACTORS IN NEUROINVASION OF ENCEPHALITIS ALPHAVIRUSES - SUMMARY THE LONG-TERM OBJECTIVE OF THIS MULTI-PI PROJECT IS TO DETERMINE HOW HOST-PATHOGEN INTERACTIONS IMPACT ENTRY, INFECTION, AND SPREAD OF ENCEPHALITIC ALPHAVIRUSES IN THE CENTRAL NERVOUS SYSTEM (CNS). THE VECTOR-BORNE NEUROTROPIC VIRUSES, VENEZUELAN, EASTERN, AND WESTERN EQUINE ENCEPHALITIS VIRUSES (VEEV, EEEV, WEEV), INVADE THE CNS AFTER SUBCUTANEOUS INOCULATION AND INITIAL INTERACTION WITH IMMUNE SENTINEL CELLS, SUCH AS MACROPHAGES AND DENDRITIC CELLS (DCS) (VEEV), OR FIBROBLASTIC, OSTEOBLASTIC AND OTHER CELL TYPES (EEEV, WEEV). BOTH EEEV AND VEEV ENTER THE BRAIN VIA THE HEMATOGENOUS ROUTE BUT ONLY VEEV IS FOUND IN OLFACTORY NEURONS. THE CNS LACKS INTRAPARENCHYMAL LYMPHOID TISSUES AND CANNOT INITIATE ADAPTIVE IMMUNE RESPONSES; THUS, IT MAINLY RELIES ON INNATE RESPONSES COMMUNICATED THROUGH LOCAL AND SYSTEMIC CYTOKINE SECRETION, WHICH MODULATE THE STATUS OF RESIDENT NEURAL CELLS AND LIMITS VIRAL NEUROINVASION AND THE EXTENT OF INFECTION. CNSRESIDENT CELLS MAY INCLUDE MULTIPLE MEMBERS OF THE NEUROVASCULAR UNIT (NVU) SUCH AS BRAIN MICROVASCULAR ENDOTHELIAL CELLS (BMECS), PERICYTES, ASTROCYTES, MICROGLIA AND NEURONS THEMSELVES THAT MAY BE INFECTED DIRECTLY OR ACTED UPON BY REGIONALLY OR SYSTEMICALLY PRODUCED CYTOKINES. VEEV IS A HIGHLY LYMPHOTROPIC VIRUS ELICITING ROBUST SERUM CYTOKINE RESPONSES AFTER PERIPHERAL INOCULATION, WHILE EEEV TROPISM IN LYMPHOID TISSUES IS HIGHLY RESTRICTED, AND SERUM CYTOKINE RESPONSES ARE MUCH LOWER, AND IN THE CASE OF TYPE I IFN, OFTEN UNDETECTABLE. IN PUBLISHED STUDIES, TWO PRIMARY VIRULENCE FACTORS FOR EEEV IN HUMAN AND MURINE MODELS DEFINED MECHANISMS THAT SUPPRESSES REPLICATION IN IMMUNE SENTINEL MYELOID CELLS AND GREATLY LIMIT INNATE IMMUNE (ESPECIALLY INTERFERON) RESPONSES TO EEEV INFECTION IN VITRO AND IN VIVO1. HOW THESE FACTORS, WHICH INCLUDE THE PRESENCE OF BINDING SITES FOR THE HEMATOPOIETIC CELL-SPECIFIC MICRORNA, MIR142-3P, IN THE EEEV 3' UNTRANSLATED REGION (UTR)1-3, AND EFFICIENT BINDING OF EEEV TO HEPARAN SULFATE (HS) RECEPTORS ON CELLS4,5, IMPACT THE DIFFERENTIAL NEUROINVASION AND CNS DISSEMINATION OF EEEV VERSUS VEEV IS UNKNOWN. SUPPORTING THE IDEA THAT DIFFERENCES IN EEEV VERSUS VEEV CYTOKINE INDUCTION MAY BE CRITICAL TO NEUROINVASION, WE FOUND THAT TYPE I IFN-DEPENDENT RESPONSES DIRECTLY REGULATE TRANSCYTOSIS, PREVENTING ALPHAVIRUS ENTRY ACROSS THE BBB AND MODULATING THE LEVEL OF INFECTION AND INJURY IN CELLS OF THE NVU6. THUS, SYSTEMIC AND LOCAL CYTOKINE RESPONSES DURING ALPHAVIRUS INFECTION INDUCE BMECS AND PERICYTES TO REGULATE VIRAL ENTRY AT THE BLOOD-BRAIN BARRIER (BBB) AND POTENTIALLY OTHER CNS SITES. THIS IS CONSISTENT WITH THE RELATIVE EXTENT OF VIRUS REPLICATION AT TERMINAL STAGES OF DISEASE AS EEEV EXHIBITS WIDESPREAD INFECTION OF NEURONS THROUGHOUT THE CNS WHILE VEEV REPLICATION IS MUCH MORE FOCAL (UNPUBLISHED). USING MUTANT VEEV AND EEEV, NOVEL VIRAL VECTORS THAT EXPRESS INDICATORS OF INFECTION (E.G., EGFP, NANOLUCIFERASE) IN VIVO, WE HAVE OBSERVED REGIONAL HETEROGENEITY IN DOMINANT SITES OF ENTRY BETWEEN THESE ALPHAVIRUSES. WITH REGARD TO BBB ENTRY, OUR STUDIES ALSO INDICATE THAT VIRAL NEUROINVASION PRECEDES BBB DISRUPTION, UTILIZING CAVEOLIN-MEDIATED TRANSCYTOSIS TO CROSS THE BBB. WE HYPOTHESIZE THAT TYPE I INTERFERON RESPONSES DIFFERENTIALLY IMPACT THE ENTRY AND INFECTION OF EEEV AND VEEV AT THE NVU VIA VIRUS-SPECIFIC INDUCTION OF REPLICATION-RESTRICTING INNATE IMMUNE RESPONSES. TO TEST THESE HYPOTHESES WE WILL: AIM 1. DEFINE ALPHAVIRUS AND HOST SPECIFIC MECHANISM IN VITRO THAT REGULATE VIRAL ENTRY AND INFECTION AT THE NVU. AIM 2: DEFINE THE IN VIVO FUNCTIONAL ROLE OF TYPE I IFN IN PROTECTION FROM ALPHAVIRUS NEUROINVASION AT THE NVU.

NEXT-GENERATION HUMANIZED MICE TO INVESTIGATE HOW TBI INTERACTS WITH GENETIC RISK FACTORS TO TRIGGER DEMENTIA.

ADVANCED DEVELOPMENT OF A THERAPEUTIC HUMANIZED ANTI-INFLAMMATORY ANTIBODY

IMPROVING OUR UNDERSTANDING OF THE NEAR-EARTH METEOROID ENVIRONMENT IS VITAL TO THE SAFETY AND SURVIVABILITY OF NASA SPACECRAFT. RISK ASSESSMENTS FOR

THE OVERARCHING GOALS OF THIS COLLABORATIVE PROGRAM ARE: 1. TO CHARACTERIZE AND MODEL QUANTITATIVELY METEOR MEASUREMENTS WITH A FOCUS ON ESTABLISHING BIASES AND THEREFORE DEFINE THE "TRUE" METEOROID ENVIRONMENT 2. ESTABLISH ESTIMATES OF UNCERTAINTIES FOR MEASURED QUANTITIES RELATED TO THE METEOROID ENVIRONMENT 3. PROVIDE OPERATIONAL SITUATIONAL AWARENESS OF THE METEOROID ENVIRONMENT.

A JOINT PROGRAM FOR METEOROID ENVIRONMENT CHARACTERIZATION AND MODELLING

MONITORING AND MODELING OF THE METEOROID ENVIRONMENT IS VITAL TO THE SAFETY AND SURVIVABILITY OF SPACECRAFT. ENGINEERING DESIGN OF SPACECRAFT TO MITI

A JOINT PROGRAM FOR METEOROID MEASUREMENT MODELLING AND MONITORING

TARGETING LUNG-DERIVED PROTEINS AS A THERAPEUTIC STRATEGY AGAINST BREAST CANCER METASTASIS

CHARACTERIZING THE AGGRESSIVENESS OF PROSTATE CANCER WITH MULTIMODALITY IMAGING

NON-INVASIVE MONITORING OF CRISPR/CAS-EDITED CHIMERIC ANTIGEN RECEPTOR T (CAR-T) CELLS WITH REPORTER GENE-BASED MAGNETIC RESONANCE IMAGING AND POSITRON EMISSION TOMOGRAPHY - PROJECT SUMMARY/ABSTRACT CLUSTERED REGULARLY INTERSPACED SHORT PALINDROMIC REPEATS/CRISPR ASSOCIATED NUCLEASE (CRISPR/CAS) GENOME EDITING HAS ENABLED SCIENTISTS TO DESIGN AND BUILD NEXT-GENERATION CELL-BASED THERAPIES. T CELL-BASED IMMUNOTHERAPIES ARE PARTICULARLY SUITABLE CRISPR/CAS TOOLS AS AFTER EDITING THEY CAN BE EXPANDED TO SUFFICIENT NUMBERS FOR USE IN HUMANS. RECENT EFFORTS HAVE SHOWN THAT PRECISE CRISPR/CAS KNOCK-IN AT THE T CELL RECEPTOR A CONSTANT (TRAC) LOCUS WITH A CHIMERIC ANTIGEN RECEPTOR (CAR) TARGETED TOWARDS THE B CELL ANTIGEN CD19 IS MORE EFFECTIVE OVER RANDOMLY INTEGRATING VIRAL-BASED ENGINEERED CAR-T CELLS IN MOUSE MODELS OF HUMAN B CELL LEUKEMIA. THEREFORE, WHILE CRISPR/CAS CELLULAR THERAPIES MAY IMPROVE PATIENT OUTCOMES, LIKE TRADITIONAL CAR- T CELL THERAPY, IT IS NOT KNOWN IF EVERY PATIENT WILL RESPOND EQUIVALENTLY OR IF SOME PATIENTS WILL BE PRONE TO OFF- TARGET, LIFE-THREATENING SIDE EFFECTS. WHAT IS NEEDED ARE TRANSLATIONALLY-RELEVANT TECHNOLOGIES THAT CAN MONITOR THE FATE OF CRISPR/CAS-EDITED CAR-T CELLS IN VIVO TO PROVIDE THE INFORMATION NEEDED TO UNDERSTAND PATIENT TREATMENT RESPONSE/NON-RESPONSE, RELAPSE AND/OR TOXICITY. THE LONG-TERM OBJECTIVE OF THIS APPLICATION IS TO DEVELOP MOLECULAR IMAGING REPORTER GENE TECHNOLOGIES FOR NON-INVASIVE VISUALIZATION OF TRAC-TARGETED CAR-T CELLS IN PATIENTS. SPECIFICALLY, WE PROPOSE TO DEVELOP NOVEL TRANSLATIONALLY-RELEVANT MAGNETIC RESONANCE IMAGING (MRI) AND POSITRON EMISSION TOMOGRAPHY (PET) REPORTER GENE ASSAYS TO NONINVASIVELY TRACK THE FATE OF TRAC-EDITED CAR-T CELLS OVER TIME AND TO VALIDATE THESE TECHNOLOGIES IN PRECLINICAL MOUSE MODELS. THE SPECIFIC AIMS OF THIS NEW APPLICATION ARE: TO COMPARE MINICIRCLE AND NON-INTEGRATING LENTIVIRAL CRISPR/CAS DONOR VECTORS FOR THEIR ABILITY TO EFFICIENTLY EDIT T CELLS AT THE TRAC LOCI WITH A CD19-TARGETED CAR (AIM 1.1); TO DEVELOP DONOR VECTORS CO-ENCODING OUR CAR, A BIOLUMINESCENCE REPORTER GENE, AND EITHER THE HUMAN MRI REPORTER ORGANIC ANION TRANSPORTER POLYPEPTIDE 1B3 (OATP1B3) OR THE PET REPORTER SODIUM IODIDE SYMPORTER (NIS), AND TO VALIDATE TRAC-EDITING AND FUNCTIONAL REPORTER EXPRESSION IN VITRO (AIM 1.2); TO EVALUATE THE SENSITIVITY OF OATP1B3-BASED MRI AND NIS-BASED PET FOR VISUALIZING CAR-T CELLS IN VIVO, AS WELL AS PERFORM LONGITUDINAL PET AND MRI OF CAR-T CELLS IN MOUSE MODELS OF B CELL MALIGNANCIES (AIM 2.1); AND FINALLY, TO DEVELOP A DUAL-REPORTER TRANSLATIONALLY-RELEVANT CRISPR/CAS SYSTEM AND EVALUATE THE ABILITY TO NONINVASIVELY MONITOR CAR-T CELLS IN MICE WITH BOTH PET AND MRI (AIM 2.2). THE SIGNIFICANCE OF THIS WORK WILL BE TO PROVIDE POWERFUL CELL TRACKING TECHNOLOGIES TO ALLOW THE FATE OF CRISPR/CAS-EDITED CAR-T CELLS, OR OTHER CRISPR/CAS-EDITED CELLULAR THERAPIES, TO BE MONITORED NONINVASIVELY IN PRECLINICAL MODELS AND PATIENTS. THIS INFORMATION WILL ALLOW MORE PRECISE MONITORING OF THESE TRANSFORMATIVE THERAPIES IN INDIVIDUAL PATIENTS TO BETTER ASSESS SAFETY AS WELL AS TO RELATE CELL BIODISTRIBUTION TO PATIENT OUTCOMES.

D18AC00035

ADVANCED DEVELOPMENT OF A THERAPEUTIC, HUMANIZED, ANTI-INFLAMMATORY ANTIBODY

MICROVASCULAR O2 DELIVERY: IMPACT OF ERYTHROCYTE-RELEASED ATP

NON-INVASIVE MONITORING OF CRISPR/CAS-EDITED CHIMERIC ANTIGEN RECEPTOR T (CAR-T) CELLS WITH REPORTER GENE-BASED MAGNETIC RESONANCE IMAGING AND POSITRON EMISSION TOMOGRAPHY

ROLE OF OVO-LIKE 1 IN THE REGULATION OF HUMAN TROPHOBLAST DIFFERENTIATION

USING GENETIC BUFFERING RELATIONSHIPS IDENTIFIED IN FISSION YEAST TO ELUCIDATE THE MOLECULAR PATHOLOGY OF TUBEROUS SCLEROSIS

ENGINEERING A CHIMERIC SERPIN AS A PREVENTIVE THERAPEUTIC FOR ISCHEMIC STROKE

CSP - INTERNATIONAL CONFERENCE ON AUDITORY CORTEX

TO FUND THE TRAVEL COSTS FOR UP TO FOUR U.S. SPEAKERS.

GRANT WILL SUPPORT U.S. RELATED CONFERENCE ENTITLED "STATES OF EMERGENCY: CRISIS PANIC AND NATION." THE CONFERENC WILL FOCUS ON HOW AMERICANS HAVE H

EXTENSIVE METERIOD OBSERVATION