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Principal Investigator: Or Shemesh
Organization: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
Fiscal Year: 2023
Award: $238,500
Funding agency: National Institute of Biomedical Imaging and Bioengineering
Abstract
Glia are supportive cells in the human brain, comprising microglia, oligodendrocytes, astrocytes, and ependymal cells.
Glia are deeply involved in diseases of the nervous system such as Alzheimer’s (AD), autism, pain, affective disorders,
and cancers. Different glial cell types play different mechanistic roles in disease formation, driven by specific genes.
Modulating glial gene expression via a process called gene therapy could thus be studied as a means of preventing
deleterious effects of glia in the brain. However, while significant progress has been made in delivering genes exclusively
to neurons, such capabilities are lacking for glia, despite their demonstrated role in disease formation, posing a critical
medical need. Although gene delivery to neurons can be achieved using viral vectors, their use to transmit genes to glia
in-vivo has been unsuccessful. Here, we propose to design a novel nonviral gene delivery vector targeting microglia or
astrocytes exclusively by bioengineering Modified RNAs (ModRNAs). ModRNAs are synthetic RNA molecules known
not to trigger an immune response and are strongly expressed in target cells. Currently, ModRNAs enable only days-
long expression, impeding long-duration medical applications and lacking cell specificity to glia types. We will engineer
glia-type-specific, ModRNAs-based constructs, GliaRNAs, as a platform for glia-exclusive gene therapy, with a
customizable expression duration. First, ModRNAs that enable robust and prolonged expression (7-14 days) will be
developed (Aim 1). For this purpose, existing ModRNA will be altered, through modifications and by inflicting random
mutations of structural components of the molecule, including CAP analog, 3’ untranslated region, coding region, 5’
untranslated region, and the poly-A tail. We will test the expression of these novel GliaRNAs in glial cultures from mice.
Next, the vector specificity will be optimized (Aim 2). We will screen for molecular manipulations that enable robust and
specific delivery of the GliaRNAs into either microglia or astrocytes (GliaRNA-vectors) and select the best gene delivery
vectors, specifically either lipid nanoparticles, antibody-lipid conjugates, or aptamers. As a proof of concept, we will use
the new GliaRNA-vector technology to express the green fluorescent protein (GFP) in either astrocytes or microglia in
mice brains. The GliaRNA-vector platform will pave the way for genetically healing and modifying different types
of glia, opening multiple therapeutic and research avenues in humans by targeting neurodegeneration, autism,
pain disorders, mood disorders, and brain cancers.
Terms: <3' Untranslated Regions><3'UTR><5' Untranslated Regions><5'UTR><AD dementia><Acute><Address><Adeno-Associated Viruses><Affect><Affective Disorders><Alzheimer Type Dementia><Alzheimer disease dementia><Alzheimer sclerosis><Alzheimer syndrome><Alzheimer's><Alzheimer's Disease><Alzheimers Dementia><Antibodies><Astrocytes><Astrocytus><Astroglia><Autism><Autistic Disorder><Binding><Bioluminescent Proteins><Biomedical Engineering><Brain><Brain Cancer><Brain Diseases><Brain Disorders><Brain Nervous System><Cancer Treatment><Cancers><Cell Body><Cell Surface Antigens><Cells><Cholesterol><Clinical><Code><Coding System><DNA Therapy><Degenerative Neurologic Diseases><Degenerative Neurologic Disorders><Dependoparvovirus><Dependovirus><Development><Disease><Disorder><Early Infantile Autism><Encephalon><Encephalon Diseases><Engineering><Ependymal Cell><Ependymocyte><Exhibits><Gene Delivery><Gene Expression><Gene Targeting><Gene Therapy Vectors><Gene Transduction Agent><Gene Transduction Vectors><Gene Transfer Clinical><Genes><Genetic Alteration><Genetic Change><Genetic Intervention><Genetic defect><Genetics-Mutagenesis><Glia><Glial Cells><Green Fluorescent Proteins><HeLa><Hela Cells><Hortega cell><Human><Image><Immune response><Immunologic Surface Markers><Immunological Surface Markers><Immunological response><Infantile Autism><Intervention><Intervention Strategies><Intracranial CNS Disorders><Intracranial Central Nervous System Disorders><Kanner's Syndrome><Knock-out><Knockout><Kolliker's reticulum><Lentivirinae><Lentivirus><Lipids><Luciferase Immunologic><Luciferases><Luminescent Proteins><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Malignant Neoplasms><Malignant Tumor><Malignant Tumor of the Brain><Malignant neoplasm of brain><Medical><Methods><Mice><Mice Mammals><Microglia><Modeling><Modern Man><Modification><Molecular><Molecular Interaction><Mood Disorders><Murine><Mus><Mutagenesis><Mutagenesis Molecular Biology><Mutation><Nerve Cells><Nerve Degeneration><Nerve Unit><Nervous System Degenerative Diseases><Nervous System Diseases><Neural Cell><Neural Degenerative Diseases><Neural degenerative Disorders><Neurocyte><Neurodegenerative Diseases><Neurodegenerative Disorders><Neuroglia><Neuroglial Cells><Neurologic Degenerative Conditions><Neurologic Disorders><Neurological Disorders><Neuron Degeneration><Neurons><Non-Polyadenylated RNA><Non-neuronal cell><Nonneuronal cell><Oligodendrocytes><Oligodendrocytus><Oligodendroglia><Oligodendroglia Cell><Optics><Pain><Pain Disorder><Painful><Pathology><Photoproteins><Play><Poly(A) Tail><Primary Senile Degenerative Dementia><Process><Proteins><RNA><RNA Gene Products><RNA Stability><Rapid screening><Reporter Genes><Research><Ribonucleic Acid><Role><Specificity><Surface Antigens><Tail><Technology><Testing><Therapeutic><Time><Transmission><Vaccination><Vascular Diseases><Vascular Disorder><Viral Vector><Virus><adeno associated virus group><analog><anti-cancer therapy><anticancer therapy><aptamer><astrocytic glia><autism spectrum disorder><autistic spectrum disorder><bio-engineered><bio-engineers><bioengineering><biological engineering><blood vessel disorder><cancer therapy><cancer-directed therapy><cell type><clinical applicability><clinical application><degenerative diseases of motor and sensory neurons><degenerative neurological diseases><delivery vector><delivery vehicle><design><designing><developmental><gene repair therapy><gene therapy><gene-based therapy><genetic therapy><genome mutation><genomic therapy><gitter cell><healing><host response><imaging><immune system response><immunogenicity><immunoresponse><in vivo><interest><interventional strategy><lipid based nanoparticle><lipid nanoparticle><mRNA Leader Sequences><malignancy><mesoglia><microglial cell><microgliocyte><neoplasm/cancer><nerve cement><nervous system disorder><neural degeneration><neurodegeneration><neurodegenerative><neurodegenerative illness><neurological degeneration><neurological disease><neuronal><neuronal degeneration><non-viral gene delivery><nonviral gene delivery><novel><optical><perivascular glial cell><prevent><preventing><primary degenerative dementia><screening><screenings><senile dementia of the Alzheimer type><site targeted delivery><social role><targeted delivery><transmission process><vascular dysfunction><vasculopathy><vector>