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Principal Investigator: Megan Culler Freeman
Organization: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
Fiscal Year: 2024
Award: $62,858
Funding agency: National Institute of Allergy and Infectious Diseases
Megan Freeman, MD, PhD, is a Pediatric Infectious Diseases (ID) Fellow at the University of Pittsburgh
Medical Center (UPMC) Children’s Hospital of Pittsburgh (CHP). She earned MD and PhD degrees at
Vanderbilt University where she investigated the cell biology and pathogenesis of coronavirus infection (Mark
Denison, MD). After clinical training in Pediatrics and Pediatric ID at CHP, she joined the lab of Carolyn Coyne,
PhD, an expert in enteroviruses and tissue barriers to viral infections. Her research focused on understanding
enterovirus D68 (EV-D68) pathogenesis, the hypothesized cause of acute flaccid myelitis (AFM), a polio-like
syndrome of paralysis in previously healthy children. Dr. Freeman used intestinal organoids and primary
bronchial epithelial cells to show that some contemporary EV-D68 isolates associated with AFM were able to
use the intestinal tract for replication in addition to the respiratory tract, suggesting an alternate infection route.
This work formed the basis for the spinal cord organoid model featured in this proposal.
Dr. Freeman will build on >10 years of experience studying viral-host interactions while gaining new
proficiencies in immunology, neuroscience, and imaging techniques which will facilitate her transition into an
independently-funded pediatrician-scientist. Dr. Freeman has assembled a multidisciplinary mentorship and
advisory team, chaired by primary mentor Dr. John Williams (respiratory virus immunity/pathogenesis) and co-
mentor Dr. Terence Dermody (viral pathogenesis/encephalitis). Additional advisory members include Dr.
Carolyn Coyne (enteroviruses/organoids), Dr. Philana Lin (immunology) and Dr. Clayton Wiley
(neuroscience/encephalitis). The committee will facilitate technique acquisition, career development, and
collaborations. The training plan incorporates coursework, technical expertise, skills in written and oral
communication, responsible conduct of research, presentation at conferences, and a timeline for seeking
additional funding. The University of Pittsburgh School of Medicine is an outstanding training environment due
to the record of scientific achievement, resources, commitment to physician scientists, and expertise.
AFM peaked biennially from 2012-2018 coincident with EV-D68 circulation, however, reasons why
contemporary EV-D68 targets the nervous system are unknown. The central hypothesis of this proposal is that
contemporary strains of EV-D68 directly infect motor neurons of the spinal cord, resulting in neuronal damage
and paralysis. Using an innovative human spinal cord organoid model, the Aims will 1) define spinal cord
replication of EV-D68, identify infected cell type(s), and discover contributions to pathogenesis, 2) define the
host cellular response of spinal cord organoids to infection with EV-D68, and 3) evaluate glutamate alterations
during infection of spinal cord organoids with EV-D68. The studies in this proposal utilize advanced tissue
modeling and imaging techniques, viral replication assays, as well as host cytokine, cellular death, and RNA
transcript analysis to understand how CNS infection with EV-D68 leads to AFM.
Terms: <0-11 years old><3-D><3-Dimensional><3D><AIDS Virus><Achievement><Achievement Attainment><Acids><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Acute Poliomyelitis><Advisory Committees><Antibodies><Assay><Basic Research><Basic Science><Bioassay><Biologic Models><Biological Assay><Biological Models><Body Tissues><Brain Inflammation><CNS Nervous System><CNS infection><CSF3><CSF3 gene><Cell Body><Cell Line><Cell Lineage><Cell model><CellLine><Cells><Cellular Tropism><Cellular biology><Cellular injury><Cellular model><Central Nervous System><Central Nervous System Infections><Central Nervous System Infectious Disease><Central Nervous System Infectious Disorder><Cerebrospinal Fluid><Cessation of life><Child><Child Youth><Childhood><Children (0-21)><Children's Hospital><Circulation><Collaborations><Communicable Diseases><Coronaviridae Infections><Coronavirus Infections><Coxsackie Viruses><Coxsackievirus><Cytolysis><Data><Death><Degenerative Neurologic Disorders><Disease><Disorder><Doctor of Philosophy><EV-68><EV-71><EV-A71><EV-D68><Echo Viruses><Echovirus><Encephalitis><Enterovirus><Enterovirus 68><Enterovirus 71><Enterovirus A71><Enterovirus D68><Enterovirus Infections><Environment><Epithelial Cells><Epithelium><Family><Foundations><Funding><G-CSF><GCSF><Genes><Genetic Differentiation><Genetic Divergence><Genetic Diversity><Genetic Drift><Genetic Variation><Glutamate Translocase><Glutamate Transport Glycoprotein><Glutamate Transporter><Glutamates><Goals><HIV><Human><Human Immunodeficiency Viruses><IFN><Imaging Procedures><Imaging Technics><Imaging Techniques><Immune><Immune response><Immunes><Immunity><Immunochemical Immunologic><Immunologic><Immunological><Immunological response><Immunologically><Immunologics><Immunology><Induced Neurons><Infection><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Inflammatory Response><Injury><Innate Immune Response><Innate Immunity><Interferons><Intestinal><Intestines><Intramuscular Injections><Investigation><Kinetics><Knowledge><L-Glutamate><LAV-HTLV-III><Left><Location><Lymphadenopathy-Associated Virus><Lysis><MGC45931><Mammalia><Mammals><Measles><Mediating><Medical center><Medulla Spinalis><Mentors><Mentorship><Mice><Mice Mammals><Model System><Modeling><Modern Man><Motor Cell><Motor Neurons><Murine><Mus><NIAID><National Institute of Allergy and Infectious Disease><Native Immunity><Natural Immunity><Nerve Cells><Nerve Transmitter Substances><Nerve Unit><Nervous System><Nervous System Degenerative Diseases><Neural Cell><Neural Degenerative Diseases><Neural degenerative Disorders><Neuraxis><Neurocyte><Neurodegenerative Diseases><Neurodegenerative Disorders><Neurologic Body System><Neurologic Degenerative Conditions><Neurologic Organ System><Neuronal Injury><Neurons><Neurosciences><Neurotransmitters><Non-Polyadenylated RNA><Non-Specific Immunity><Nonspecific Immunity><Organoids><Palsy><Paralysed><Pathogenesis><Pathway interactions><Patients><Pediatric Hospitals><Pediatrics><Ph.D.><PhD><Physicians><Physiologic><Physiological><Plegia><Polio><Poliomyelitis><Pulmonary Body System><Pulmonary Organ System><RNA><RNA Gene Products><Research><Research Resources><Residual><Residual state><Resources><Respiratory System><Respiratory Tracts><Respiratory tract structure><Ribonucleic Acid><Route><Rubeola><Sampling><Scientist><Site><Specificity><Spinal><Spinal Cord><Stomach><Strains Cell Lines><Syndrome><Task Forces><Technical Expertise><Techniques><Temperature><Tissue Model><Tissue imaging><Tissues><Training><Transcript><Universities><Viral><Viral Diseases><Viral Pathogenesis><Virus><Virus Diseases><Virus Replication><Virus-HIV><Work><Writing><acute flaccid myelitis><advisory team><bowel><bronchial epithelium><career development><cell biology><cell damage><cell injury><cell type><cellular damage><cerebral spinal fluid><clinical training><conference><convention><cultured cell line><cytokine><damage to cells><degenerative diseases of motor and sensory neurons><degenerative neurological diseases><disease phenotype><excitotoxic><excitotoxicity><experience><experiment><experimental research><experimental study><experiments><gastric><glutamatergic><host response><human model><human tissue><iNeuron><immune system response><immunoresponse><injuries><injury to cells><innovate><innovation><innovative><intramuscular drug administration><kids><medical college><medical schools><member><model of human><morbilli><motoneuron><multidisciplinary><neonatal mice><neural mechanism><neurodegenerative illness><neuromechanism><neuron injury><neuronal><novel><novel virus><oral communication><paralysis><paralytic><pathway><pediatric><pediatric scientist><pediatrician physician scientist><pediatrician scientist><prevent><preventing><programs><respiratory><respiratory virus><response><responsible research conduct><school of medicine><skills><spinal fluid><summit><symposia><symposium><technical skills><therapeutic target><three dimensional><timeline><viral infection><viral multiplication><viral replication><virus infection><virus multiplication><virus pathogenesis><virus-induced disease><youngster>