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Principal Investigator: Yeh-Hsing Lao
Organization: STATE UNIVERSITY OF NEW YORK AT BUFFALO
Fiscal Year: 2023
Award: $442,000
Funding agency: National Institute of Neurological Disorders and Stroke
Nanoformulated CRISPR Ribonucleoproteins for Ultrasound-facilitated Brain Gene Editing
Abstract
Emerging CRISPR technologies provide new opportunities to advance gene therapy in treating many
intractable genetic diseases, including neuronal degeneration disorders. Given the failures of clinical trials in
treating Alzheimer's disease through directly targeting amyloid β and tau, there is an unmet need to develop a
different strategy in this space, and gene editing technologies may be of great potential. However, one key barrier
in developing CRISPR therapeutics is the brain delivery of CRISPR components. Viral vectors could be effective,
but the use of these vectors could potentially raise the concerns in immunogenicity and toxicity, which may lead
to severe adverse effects. Conventional nonviral systems, in contrast, could be safer but significantly less
effective, possibly due to the suboptimal size, which limits their transport to the target brain region. In light of
these challenges, we propose to explore the feasibility of screening more transport-favorable, effective nonviral
carriers for brain gene editing to tackle Alzheimer's disease. Different from the conventional nanoparticle designs,
we will first create a large nanoformulated CRISPR/Cas9 ribonucleoprotein library through split-and-pool lipid
coating and optimize the focused ultrasound (FUS)-mediated blood-brain barrier opening to screen all the
possible lipid compositions (Aim 1). Compared with the conventional nanoparticle formulations, direct lipid
coating may generate smaller and more transport-favorable “nano editors.” By barcoding each lipid in each split-
and-pool round, all the nanoformulated Cas9 ribonucleoproteins can be screened directly in the same animal,
which minimizes the variations from animals and operations. Our preliminary studies with a small set of
nanoformulations in different models have demonstrated the feasibility and reproducibility of our screening
approach. Once having the most potent lipid composition, we will validate its gene editing performance and
therapeutic efficacy in both reporter and Alzheimer’s mouse models (Aim 2). Our previous efforts in developing
FUS delivery for viral brain gene editing have helped us established the capability and all the pipelines needed
for editing performance validations. In this proposed research, we aim to expand the CRISPR delivery toolkits
from viral to nonviral systems and to explore the potential of nonviral CRISPR gene editing for treating
Alzheimer’s disease. The discoveries and findings will help us gain enough supports for larger, potentially IND-
enabling studies.
Terms: <AAV vector><AD dementia><AD model><Adverse effects><Alzheimer Type Dementia><Alzheimer beta-Protein><Alzheimer disease dementia><Alzheimer sclerosis><Alzheimer syndrome><Alzheimer's><Alzheimer's Amyloid beta-Protein><Alzheimer's Disease><Alzheimer's amyloid><Alzheimer's disease model><Alzheimers Dementia><Amyloid Alzheimer's Dementia Amyloid Protein><Amyloid Beta-Peptide><Amyloid Protein A4><Amyloid beta-Protein><Amyloid β><Amyloid β-Peptide><Amyloid β-Protein><Animal Model><Animal Models and Related Studies><Animal Testing><Animals><Aβ><BBB crossing><Bar Codes><Biologic Products><Biological Agent><Biological Products><Blood - brain barrier anatomy><Blood-Brain Barrier><Brain><Brain Nervous System><Brain region><CRISPR><CRISPR approach><CRISPR based approach><CRISPR based therapeutics><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR therapeutics><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas system><CRISPR/Cas technology><CRISPR/Cas therapeutics><CRISPR/Cas9><CRISPR/Cas9 technology><CRISPR/Cas9 therapeutics><CRISPR/Cas9 therapy><Cas nuclease technology><Cas9 based therapeutics><Charge><Clinical Trials><Closure by Ligation><Clustered Regularly Interspaced Short Palindromic Repeats><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats based therapeutics><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Clustered Regularly Interspaced Short Palindromic Repeats therapeutics><DNA><DNA Therapy><Deoxyribonucleic Acid><Development><Diffusion><Disease><Disorder><Encapsulated><Encephalon><Exploratory/Developmental Grant for Diagnostic Cancer Imaging><Failure><Focused Ultrasound><Formulation><Gene Transfer Clinical><Genes><Genetic Diseases><Genetic Intervention><Guide RNA><Hemato-Encephalic Barrier><Libraries><Ligase><Ligase Gene><Ligation><Light><Lipids><Liposomal><Liposomes><Liver><MT-bound tau><Mediating><Methods><Mice><Mice Mammals><Modeling><Mouse Strains><Murine><Mus><NIH><NIMH><NINDS><National Institute of Mental Health><National Institute of Neurological Diseases and Stroke><National Institute of Neurological Disorders and Stroke><National Institutes of Health><Nature><Nerve Cells><Nerve Degeneration><Nerve Unit><Neural Cell><Neurocyte><Neuron Degeneration><Neurons><Nucleic Acids><Performance><Photoradiation><Primary Senile Degenerative Dementia><R21 Award><Race><Races><Reporter><Reproducibility><Research><Ribonucleoproteins><Running><Safety><Sampling><Surface><Synthetases><System><TIL4><TLR2><TLR2 gene><TLR2 receptor><Techniques><Technology><Time><Toll-Like Receptor 2><Toll/Interleukin 1 Receptor-Like 4><Toll/Interleukin 1 Receptor-Like 4 Gene><Toll/Interleukin 1 Receptor-Like Protein 4><Toxic effect><Toxicities><Transfection><Transgenic Organisms><Translations><Treatment Efficacy><United States National Institutes of Health><Validation><Variant><Variation><Viral><Viral Genes><Viral Vector><a beta peptide><abeta><adeno-associated viral vector><adeno-associated virus vector><alzheimer model><amyloid beta><amyloid-b protein><barcode><behavior test><behavioral test><beta amyloid fibril><biologics><biopharmaceutical><biotherapeutic agent><blood-brain barrier crossing><bloodbrain barrier><bloodbrain barrier crossing><brain tissue><design><designing><determine efficacy><developmental><diffused><diffuses><diffusing><diffusions><efficacy analysis><efficacy assessment><efficacy determination><efficacy evaluation><efficacy examination><evaluate efficacy><examine efficacy><experience><gRNA><gene repair therapy><gene therapy><gene-based therapy><genetic condition><genetic disorder><genetic therapy><genomic therapy><hepatic body system><hepatic organ system><immunogenicity><improved><intervention efficacy><microtubule bound tau><microtubule-bound tau><model of animal><mouse model><murine model><nano><nano formulation><nano particle><nano-sized particle><nanoformulation><nanoparticle><nanosized particle><neural degeneration><neurodegeneration><neurodegenerative><neurological degeneration><neuronal><neuronal degeneration><operation><operations><post-doctoral training><postdoctoral training><primary degenerative dementia><racial><racial background><racial origin><screening><screenings><senile dementia of the Alzheimer type><soluble amyloid precursor protein><somatic cell gene editing><somatic cell genome editing><somatic gene editing><somatic genome editing><tau><tau Proteins><tau factor><therapeutic efficacy><therapy efficacy><transgenic><translation><ultrasound><validations><vector><τ Proteins>