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Principal Investigator: David Wang
Organization: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
Fiscal Year: 2022
Award: $892,381
Funding agency: National Institute of Neurological Disorders and Stroke
Specific aim 1: To develop in vitro 3D brain organoid models derived from human adult peripheral CD34+ cells to study neural development and degeneration and infectious diseases involving human brain. We expended our capability to derive 3D neural organoids to mimic human brain. Using improved clearance techniques and advanced confocal imaging system, we confirmed that pericytes can be produced along endothelial cells and neural cells in the 3D organoid spontaneously with sequential induction media, making the model perfect for virus infection studies, as pericytes have been reported as a main target by viruses including SARS-CoV2. In another 3D model with incorporating CD34 cells, we treated them with cytokines to promote microglial differentiation and used scRNA-Seq analysis to study the resulting microglial cells. We found that although the treatment of cytokines is not necessary for producing microglia, it did increase the numbers of microglia compared to organoids without cytokine stimulation. When the organoids were treated with LPS, which induced inflammatory reactions by increasing transcriptions of certain microglial genes in the organoids, indicating the differentiated microglia in the organoids are functional. A manuscript is prepared for publication based on the results.
Specific aim 2: To study the roles of HERV-K on brain development and tumorigenicity. We supported Dr. Ashish Shah on studying the mechanism of HERV-K on the pathogenesis of glioma. We transfected astroglia with plasmid containing HERV-K and detected higher level of gene transcription responsible for stemness. This result indicates that marker for stem cells is activated by HERV-K and maybe the mechanism underlying its effect on tumorigenesis. We are further studying HERV-K effect in cell proliferation and tumorigenesis using 3D models.
Specific aim 3: To study the association of HERV-K and ALS. We differentiated motor neurons from four sets of C9orf72 and control iPSCs and did RNA-Seq analysis. The results showed top differential pathways were tumorigenesis related. As we have found that in C9orf72 motor neurons, there was increased DNA repairing element followed by HERV-K activation, this result imply that DNA damaging events in motor neurons leads to HERV-K Env activation, which may reverse the neurons to a status more similar to stem or tumor cells, causing motor neuron malfunction.
Specific aim 4: To facilitate research and therapeutic developments for neurological disorders using our models and methods. We are continuing to collaborate with other investigators by providing material support and technique trainings of the iNSC/iPSC generation and 3D modeling. We are working with Dr. In-Hong Yang in UNC to develop protocol to culture motor neurons in a microfluidic chamber. We collaborated with Dr. Kousa from National Childrens Hospital to study the pathogenesis of ZIKA virus on human brain development. We derived neural stem cells from targeting and control iPSC cells and found ZIKA virus infection was enhanced significantly by a target malfunction. We also helped Dr. Farinaz Safavi to generate 3D brain organoids from iPSCs with an mutative gene. Our result showed the mutation decreased the growth of the derived 3D organoids significantly compared to corrected iPSCs.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><21+ years old><3-D><3-D modeling><3-Dimensional><3D><3D modeling><Adult><Adult Human><Adventitial Cell><Assay><Astrocytes><Astrocytus><Astroglia><Bioassay><Biologic Assays><Biological Assay><Brain><Brain Nervous System><C9ORF72><CD34><CD34 gene><COVID-19 virus><COVID19 virus><Cell Body><Cell Growth in Number><Cell Multiplication><Cell Proliferation><Cells><Cellular Proliferation><Children's Hospital><Co-culture><CoV-2><CoV2><Cocultivation><Coculture><Coculture Techniques><Communicable Diseases><DNA Damage><DNA Damage Repair><DNA Injury><DNA Repair><Development><Development and Research><Diagnosis><Disease><Disorder><ERVs><EXTMR><Elements><Encephalon><Endogenous Retroviruses><Endothelial Cells><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Event><Extramural><Extramural Activities><Gene Alteration><Gene Mutation><Gene Transcription><Generalized Growth><Generations><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic Transcription><Genetic defect><Genetic study><Glial Cell Tumors><Glial Neoplasm><Glial Tumor><Glioma><Goals><Growth><HERVs><HPCA1><Hortega cell><Human><Human Endogenous Retroviruses><In Vitro><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Inflammation><Inflammatory><Investigators><Manuscripts><Methods><Microfluidics><Microglia><Modeling><Modern Man><Molecular Target><Motor Cell><Motor Neurons><Mutation><NIH><NINDS><National Institute of Neurological Diseases and Stroke><National Institute of Neurological Disorders and Stroke><National Institutes of Health><Nerve Cells><Nerve Degeneration><Nerve Unit><Nervous System Diseases><Neural Cell><Neural Development><Neural Stem Cell><Neurocyte><Neurodevelopmental Disorder><Neuroglial Neoplasm><Neuroglial Tumor><Neurologic Disorders><Neurological Development Disorder><Neurological Disorders><Neuron Degeneration><Neurons><Oncogenesis><Organoids><Pathogenesis><Pathway interactions><Patients><Pediatric Hospitals><Pericapillary Cell><Pericytes><Peripheral><Perivascular Cell><Plasmids><Progenitor Cells><Protocol><Protocols documentation><Publications><R & D><R&D><RNA Expression><RNA Seq><RNA sequencing><RNAseq><Reaction><Reporting><Research Personnel><Researchers><Role><Rouget Cells><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV2><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Sampling><Scientific Publication><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome related corona virus 2><Techniques><Therapeutic Intervention><Tissue Growth><Training Technics><Training Technique><Transcription><Tumor Cell><Tumorigenicity><United States National Institutes of Health><Unscheduled DNA Synthesis><Viral Diseases><Virus><Virus Diseases><Wuhan coronavirus><Yang><ZIKV><ZIKV infection><ZIKV positive><Zika Virus><Zika virus infection><adulthood><astrocytic glia><base><cell type><chromosome 9 open reading frame 72><confocal imaging><coronavirus disease 2019 virus><coronavirus disease-19 virus><cytokine><developmental><genome mutation><gitter cell><glial-derived tumor><hCoV19><iPS><iPSC><iPSCs><imaging system><immunoneurology><improved><induced pluripotent stem cell><inducible pluripotent stem cell><intervention therapy><mesoglia><microglial cell><microgliocyte><motoneuron><nCoV2><neoplastic cell><nerve stem cell><nervous system disorder><neural><neural degeneration><neural model><neural precursor><neural precursor cell><neural progenitor><neural progenitor cells><neurodegeneration><neurodegenerative><neurodevelopment><neurogenesis><neuroglia neoplasm><neuroglia tumor><neuroimmunology><neurological degeneration><neurological disease><neuron progenitors><neuron toxicity><neuronal><neuronal degeneration><neuronal progenitor><neuronal progenitor cells><neuronal stem cells><neuronal toxicity><neuroprogenitor><neurotoxicity><novel><ontogeny><pathway><perivascular glial cell><pre-clinical study><preclinical study><relating to nervous system><research and development><scRNA-seq><screening><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><social role><stem cell based model><stem cell biomarkers><stem cell derived model><stem cell markers><stem cell model><stem cells><stemness><therapeutic agent development><therapeutic development><three dimensional><three-dimensional modeling><transcriptome sequencing><translational neuroscience><tumorigenesis><viral infection><virus infection><virus-induced disease><zika infection><zika viral infection><zikav><µfluidic>