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Principal Investigator: Bowen Li
Organization: UNIVERSITY OF TORONTO
Fiscal Year: 2024
Award: $270,000
Funding agency: National Heart Lung and Blood Institute
PROJECT SUMMARY
Cystic fibrosis (CF) is a debilitating genetic disorder that predominantly impacts the lungs and pancreas due to
mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Specifically, nonsense
mutations, which produce truncated, non-functional proteins, account for approximately 11% of all CF cases.
Currently, these mutations lack targeted, effective therapeutic solutions. Our research endeavors to address this
unmet clinical need by harnessing the capabilities of adenine base editors (ABEs) to correct premature
termination codons (PTCs) through translational readthrough, thereby restoring functional protein expression.
To facilitate efficient delivery, we aim to formulate lipid nanoparticles (LNPs) specifically engineered for
pulmonary ABE delivery. The project is segmented into three pivotal objectives: 1) the chemical refinement of
RNA-encoded ABEs to achieve optimal on-target gene-editing; 2) the development and fine-tuning of LNP
formulations to overcome challenges intrinsic to pulmonary delivery; and 3) comprehensive in vivo validation
using a CFTR nonsense mutation mouse model to assess both therapeutic efficacy and safety profiles. By
focusing on nanoparticle-mediated lung cell engineering, our approach lays the foundation for a groundbreaking,
non-viral base editing platform that could revolutionize treatment protocols for CF and other diseases stemming
from PTCs. Employing cutting-edge methodologies, including the optimization of mRNA sequences encoding
ABEs, gRNA design, and novel four-component combinatorial lipid chemistry, our project has the potential to
substantially advance genomic editor delivery to the pulmonary system. These innovations could reconfigure
therapeutic strategies for CF and other genetic diseases caused by nonsense mutations. The feasibility of this
high-impact work is reinforced by our lab's established expertise in these technologies, making it a viable
candidate for significantly advancing the nonviral delivery of genomic medicines and thereby enhancing the
therapeutic options for a wide array of lung diseases.
Terms: <1H-Purin-6-amine><3D cell culture><3D culture><Address><Adenine><Air><American><Assay><Bioassay><Biological Assay><CF patients><CFTR><CFTR Protein><Cell Culture Techniques><Cell Line><CellLine><Chemicals><Clinical><Combinatorial Synthesis><Cystic Fibrosis><Cystic Fibrosis Transmembrane Conductance Regulator><DNA Therapy><DNA editor><Development><Disease><Disorder><Encapsulated><Endosomes><Engineering><Epithelial Cells><Event><Formulation><Foundations><Future><Gel><Gene Transcription><Gene Transfer Clinical><Gene variant><Generations><Genes><Genetic Alteration><Genetic Change><Genetic Diseases><Genetic Intervention><Genetic Transcription><Genetic defect><Genomic medicine><Genomics><Guide RNA><Health Sciences><Hereditary Disease><Human><In Vitro><In vivo analysis><Inborn Genetic Diseases><Inherited disorder><Libraries><Lipid Chemistry><Lipids><Liquid substance><Lung><Lung Diseases><Lung Respiratory System><Mediating><Medical><Messenger RNA><Methodology><Mice><Mice Mammals><Modern Man><Molecular><Mucous body substance><Mucoviscidosis><Mucus><Murine><Mus><Mutation><Nasal><Nasal Passages Nose><Nebulizer><Non-Polyadenylated RNA><Nonsense Mutation><Nose><Other Genetics><Outcome><Pancreas><Pancreatic><Patient Care><Patient Care Delivery><Patients><Penetration><Performance><Process><Proteins><Pulmonary Diseases><Pulmonary Disorder><RNA><RNA Expression><RNA Gene Products><RNA Sequences><Receptosomes><Recovery of Function><Regulator Genes><Research><Respiratory System, Nose, Nasal Passages><Ribonucleic Acid><Risk><Safety><Stop Codon><Strains Cell Lines><System><Techniques><Technology><Termination Codon><Terminator Codon><Therapeutic><Transcription><Transcriptional Regulatory Elements><Translation Stop Signal><Treatment Efficacy><Treatment Protocols><Treatment Regimen><Treatment Schedule><Validation><Variant><Variation><Vitamin B4><Work><allele variant><allelic variant><base editing><base editor><care for patients><care of patients><caring for patients><cell culture><cell cultures><cell engineering><cellular engineering><combinatorial><cultured cell line><cystic fibrosis patients><cystic fibrosis transmembrane regulator><design><designing><determine efficacy><developmental><disease model><disease of the lung><disorder model><disorder of the lung><dosage><effective therapy><effective treatment><efficacy analysis><efficacy assessment><efficacy determination><efficacy evaluation><efficacy examination><entire genome><evaluate efficacy><examine efficacy><fluid><full genome><functional recovery><functional restoration><gRNA><gene editor><gene repair therapy><gene therapy><gene-based therapy><genetic condition><genetic disorder><genetic therapy><genetic variant><genome editor><genome mutation><genome sequencing><genomic therapy><genomic variant><hereditary disorder><heritable disorder><in vivo><in vivo evaluation><in vivo testing><inborn error><individuals with CF><individuals with cystic fibrosis><inherited diseases><inherited genetic disease><inherited genetic disorder><innovate><innovation><innovative><intervention efficacy><lipid based nanoparticle><lipid nanoparticle><liquid><lung disorder><mRNA><mouse model><mucous><murine model><nano particle><nano-sized particle><nanoparticle><nanosized particle><nebulization><nebulize><non-sense mutation><novel><patients with CF><patients with cystic fibrosis><premature><prematurity><process optimization><protein expression><pulmonary><regulatory gene><restore function><restore functionality><restore lost function><risk mitigation><safety assessment><screening><screenings><stem><therapeutic efficacy><therapeutically effective><therapy efficacy><three dimensional cell culture><trans acting element><uptake><validations><whole genome>