Investigation of enterovirus D68 pathogenesis in the human spinal cord

肠道病毒 D68 在人脊髓中发病机制的研究

• 批准号: 10506843
• 负责人: Megan Culler Freeman
• 金额: $ 15.46万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-06 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10506843
• 关键词: 3-Dimensional; Achievement; Acids; Advisory Committees; Antibodies; Basic Science; Biological Assay; Biological Models; Blood Circulation; CSF3 gene; Cell Lineage; Cell model; Cells; Cellular Tropism; Cellular biology; Central Nervous System Infections; Cerebrospinal Fluid; Cessation of life; Child; Childhood; Clinical; Collaborations; Communicable Diseases; Coronavirus Infections; Coxsackie Viruses; Cytolysis; Data; Disease; Doctor of Philosophy; Echo Viruses; Encephalitis; Enterovirus; Enterovirus 68; Enterovirus Infections; Environment; Epithelial; Epithelial Cells; Family; Foundations; Funding; Genes; Genetic Drift; Genetic Variation; Glutamate Transporter; Glutamates; Goals; Human; Imaging Techniques; Immune; Immune response; Immunity; Immunologics; Immunology; Infection; Inflammatory Response; Injury; Innate Immune Response; Interferons; Intestines; Intramuscular; Investigation; Kinetics; Knowledge; Lead; Left; Location; Mammals; Mediating; Medical center; Mentors; Mentorship; Modeling; Motor Neurons; Mus; National Institute of Allergy and Infectious Disease; Natural Immunity; Nervous system structure; Neuraxis; Neurodegenerative Disorders; Neuronal Injury; Neurons; Neurosciences; Neurotransmitters; Organoids; Paralysed; Pathogenesis; Pathway interactions; Patients; Pediatric Hospitals; Pediatrics; Physicians; Physiological; Poliomyelitis; RNA; Research; Residual state; Resources; Respiratory System; Route; Sampling; Scientist; Site; Specificity; Spinal; Spinal Cord; Stomach; Syndrome; Technical Expertise; Techniques; Temperature; TimeLine; Tissue Model; Tissue imaging; Tissues; Training; Transcript; Universities; Viral; Viral Pathogenesis; Virus; Virus Diseases; Virus Replication; Work; acute flaccid myelitis; bronchial epithelium; career development; cell injury; cell type; cytokine; disease phenotype; excitotoxicity; experience; experimental study; human model; human tissue; innovation; medical schools; member; multidisciplinary; neonatal mice; neuromechanism; novel; novel virus; oral communication; pediatrician; prevent; programs; respiratory; respiratory virus; response; responsible research conduct; skills; symposium; therapeutic target

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.

Megan Freeman，医学博士，匹兹堡大学儿科传染病（ID）研究员