Document text
Principal Investigator: Rohan Palanki
Organization: UNIVERSITY OF PENNSYLVANIA
Fiscal Year: 2024
Award: $36,681
Funding agency: National Heart Lung and Blood Institute
PROJECT SUMMARY
Congenital lung diseases, such as inherited surfactant protein syndromes, cystic fibrosis, and
alpha-1 antitrypsin deficiency, are a significant source of pediatric morbidity and mortality.
Treatment options for neonatal patients with lung disorders that present with respiratory failure
are limited to palliative care or pediatric lung transplant. As such, there is a clear clinical demand
for new therapies that allow for early correction of congenital lung diseases to reduce pediatric
morbidity and mortality. Recent advances in gene editing technologies, such as CRISPR-Cas9
systems, have unlocked the potential to correct pathogenic mutations and thereby treat congenital
disorders at their source. Performing gene editing in utero offers the added benefits of reversing
genetic abnormalities prior to the transition to postnatal life, when pulmonary function becomes
essential, and harnessing normal developmental properties of the fetus for more efficient
correction. Traditionally, viral vectors have been used to study in utero gene therapy in animal
models. Although these studies are encouraging, discovery of alternative, potentially safer,
delivery vehicles will advance the field toward clinical translation. Thus, this proposal aims to
investigate the potential of ionizable lipid nanoparticles (LNPs), a promising non-viral delivery
platform, for nucleic acid delivery to the mouse fetal lung. Fetal lung optimized lipid nanoparticles
(FLO-LNPs) will be generated through a multi-stage optimization scheme. In Aim 1, a diverse
library of 24 ionizable lipid structures will be screened to identify the ionizable lipid that best
delivers mRNA to the fetal lung. In Aim 2, LNP formulations will be optimized using a Design of
Experiments scheme for minimal toxicity and maximal delivery of a CRISPR-Cas9 systems in
mouse precision cut lung slices. In Aim 3, the optimized LNP formulation will be modified via
antibody conjugation and tested for cell-specific targeting in the fetal mouse lung. Ultimately, this
proposal – conducted as an interdisciplinary project between sponsors in the Department of
Bioengineering, Perelman School of Medicine, and Children’s Hospital of Philadelphia at the
University of Pennsylvania – will allow for the development of a novel LNP delivery platform that
can be applied in subsequent work to deliver in utero gene therapies for congenital lung disease.
Terms: <AAT deficiency><Acceleration><Affect><After Care><After-Treatment><Aftercare><Airway failure><Alamar Blue><AlamarBlue><Animal Model><Animal Models and Related Studies><Animals><Antibodies><Assay><BALB C Mouse><BALB/c><Binding><Bioassay><Biocompatible Materials><Biological Assay><Bioluminescence><Biomaterials><Biomedical Engineering><Birth><Blood><Blood Circulation><Blood Reticuloendothelial System><Bloodstream><C fetus><C. fetus><CD54 Antigens><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><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 technology><CRISPR/Cas9><CRISPR/Cas9 technology><Campylobacter fetus><Cas nuclease technology><Cell Body><Cells><Charge><Childhood><Children's Hospital><Cholesterol><Chromosomal, Gene, or Protein Abnormality><Chronologic Fetal Maturity><Clinical><Clinical Medical Sciences><Clinical Medicine><Clinical Treatment Moab><Clustered Regularly Interspaced Short Palindromic Repeats approach><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><Complex><Congenital Disorders><Cystic Fibrosis><Cytogenetic or Molecular Genetic Abnormality><DNA Therapy><Development><Disease><Disorder><Dose><Encapsulated><Endocytosis><Environment><Epithelial Cells><Epithelium><Evaluation><Experimental Designs><FDA approved><Fetal Age><Fetal Lung><Fetus><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Fluorescence><Formulation><Gene Transfer Clinical><Genes><Genetic Abnormality><Genetic Alteration><Genetic Change><Genetic Intervention><Genetic defect><Gestational Age><Guide RNA><Harvest><Hereditary><Hospital Admission><Hospitalization><Hour><ICAM-1><IVIS SpectrumCT><IVIS imaging><IVIS optical imaging><IVIS spectral imaging><IVIS spectrum><IVIS system><Immune system><Inbred BALB C Mice><Inherited><Injections><Intercellular adhesion molecule 1><Lead><Libraries><Life><Lipid Chemistry><Lipids><Literature><Luciferase Immunologic><Luciferases><Lung><Lung Diseases><Lung Grafting><Lung Respiratory System><Lung Transplantation><Lytotoxicity><Macrogols><Maleimides><Mediating><Mentorship><Messenger RNA><Mice><Mice Mammals><Microfluidics><Modeling><Molecular Abnormality><Molecular Interaction><Monoclonal Antibodies><Morbidity><Morbidity - disease rate><Mucoviscidosis><Murine><Mus><Mutation><Non-Polyadenylated RNA><Nucleic Acids><Palliative Care><Palliative Therapy><Palliative Treatment><Parturition><Pathogenicity><Pb element><Pediatric Hospitals><Pennsylvania><Phenotype><Philadelphia><Phosphatides><Phospholipids><Polyethylene Glycols><Polyethylene Oxide><Polyethyleneoxide><Polyoxyethylenes><Progenitor Cells><Property><Protein C Deficiency><Proteins><Pulmonary Diseases><Pulmonary Disorder><Pulmonary Graft><Pulmonary Surfactant Protein C><Pulmonary Surfactant-Associated Protein C><Pulmonary Surfactant-Associated Protein SP-C><Pulmonary Transplant><Pulmonary Transplantation><RNA><RNA Gene Products><RNA delivery><Reaction><Reporter><Respiration Disorders><Respiratory Disease><Respiratory Disorder><Respiratory Failure><Respiratory System Disease><Respiratory System Disorder><Respiratory physiology><Ribonucleic Acid><SP-C peptide><SP-C protein><Safety><Scheme><Slice><Source><Staining method><Stains><Structure><Surface><Surfactant Polypeptide SP-C><Syndrome><System><Testing><Toxic effect><Toxicities><Trachea><Trachea Proper><Training><Transfection><Universities><Vibrio fetus><Viral Vector><Work><a1-antitrypsin deficiency><adenoviral mediated><adenovirus-mediated><alpha 1-Antitrypsin Deficiency><alpha-1-anti-trypsin deficiency><alpha1-antitrypsin deficiency><antibody conjugate><bio-engineered><bio-engineers><biocompatibility><bioengineering><biological engineering><biological material><biomaterial compatibility><breathing disorder><cell type><clinical translation><clinically translatable><comfort care><conference><convention><curative intervention><curative therapeutic><curative therapy><curative treatments><cytokine><cytotoxicity><deliver mRNA><deliver messenger RNA><delivery system for mRNA><delivery vector><delivery vehicle><design><designing><developmental><disease of the lung><disorder of the lung><experience><fetal><flow cytophotometry><gRNA><gene editing platform><gene editing system><gene editing technology><gene editing tools><gene repair therapy><gene therapy><gene-based therapy><gene-editing toolkit><genetic therapy><genome editing><genome mutation><genomic editing><genomic therapy><genotoxicity><heavy metal Pb><heavy metal lead><in utero><in vivo><in vivo imaging system><lead optimization><lipid based nanoparticle><lipid nanoparticle><lung disorder><lung function><lung transplant><mAbs><mRNA><mRNA delivery><medical college><medical schools><messenger RNA delivery><model of animal><molecular aberrations><monoclonal Abs><mortality><mouse model><murine model><nano particle delivery><nanoparticle delivered><nanoparticle delivery><neonatal patient><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation><next generation therapeutics><novel><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><nucleic acid delivery><pediatric><post treatment><postnatal><pre-clinical><preclinical><pulmonary><pulmonary function><respiratory dysfunction><respiratory function><reverse genetics><school of medicine><stem cells><summit><surfactant><surfactant protein C><symposia><symposium><uptake><windpipe><µfluidic><α-1 anti-trypsin deficiency><α-1-antitrypsin deficiency><α1-Antitrypsin Deficiency>