Ionizable Lipid Nanoparticles for Fetal Lung Targeting

用于胎儿肺靶向的可电离脂质纳米颗粒

基本信息

批准号：
10708763
负责人：
Rohan Palanki
金额：
$ 5.27万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10708763
关键词：
Acceleration Affect Aftercare AlamarBlue Animal Model Animals Antibodies Binding Biocompatible Materials Biological Assay Bioluminescence Biomedical Engineering Birth Blood Blood Circulation CRISPR/Cas technology Campylobacter fetus Cells Charge Childhood Cholesterol Clinical Clinical Medicine Complex Congenital Disorders Cystic Fibrosis Development Disease Dose Encapsulated Endocytosis Environment Epithelial Cells Epithelium Evaluation Experimental Designs FDA approved Fetal Lung Fetus Flow Cytometry Fluorescence Formulation Genes Gestational Age Guide RNA Harvest Hospitalization Hour Immune system Inbred BALB C Mice Inherited Injections Intercellular adhesion molecule 1 Lead Libraries Life Lipid Chemistry Lipids Literature Luciferases Lung Lung Transplantation Lung diseases Maleimides Mediating Mentorship Messenger RNA Microfluidics Modeling Molecular Abnormality Monoclonal Antibodies Morbidity - disease rate Mus Mutation Nucleic Acids Palliative Care Pathogenicity Pediatric Hospitals Pennsylvania Phenotype Philadelphia Phospholipids Polyethylene Glycols Property Protein C Deficiency Proteins Pulmonary Surfactant-Associated Protein C RNA RNA delivery Reaction Reporter Respiration Disorders Respiratory Disease Respiratory Failure Respiratory physiology Safety Scheme Slice Source Stains Structure Surface Syndrome System Technology Testing Toxic effect Trachea Training Transfection Universities Viral Vector Work adenoviral mediated alpha 1-Antitrypsin Deficiency antibody conjugate biomaterial compatibility cell type clinical translation curative treatments cytokine cytotoxicity delivery vehicle design experience fetal gene therapy genome editing genotoxicity in utero in vivo in vivo imaging system lead optimization lipid nanoparticle medical schools mortality mouse model nanoparticle delivery neonatal patient next generation novel novel therapeutics nucleic acid delivery postnatal pre-clinical pulmonary function reverse genetics stem cells surfactant symposium uptake

项目摘要

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.

项目概要先天性肺部疾病，例如遗传性表面活性蛋白综合征、囊性纤维化和 α-1抗胰蛋白酶缺乏症是儿科发病和死亡的重要根源。患有呼吸衰竭的肺部疾病新生儿患者的治疗选择仅限于姑息治疗或儿科肺移植，因此存在明确的临床需求。寻找能够及早纠正先天性肺部疾病的新疗法，以减少儿科疾病的发生基因编辑技术（例如 CRISPR-Cas9）的最新进展。系统，已经释放了纠正致病突变的潜力，从而治疗先天性在子宫内进行基因编辑可以带来逆转疾病的额外好处。过渡到产后生活之前的遗传异常，此时肺功能变得至关重要，并利用胎儿的正常发育特性来提高效率传统上，病毒载体已用于动物子宫基因治疗的研究。尽管这些研究令人鼓舞，但发现了可能更安全的替代方案，因此，该提案旨在推动该领域的临床转化。研究可电离纳米脂质颗粒（LNP）的潜力，这是一种有前途的非病毒传递平台，用于将核酸递送至小鼠胎肺优化的脂质纳米颗粒。（FLO-LNP）将通过多阶段优化方案生成。目标 1 是一种多样化的方案。将筛选 24 个可电离脂质结构库，以确定最适合的可电离脂质在目标 2 中，LNP 配方将使用以下设计进行优化。 CRISPR-Cas9 系统毒性最小和最大递送的实验方案在目标 3 中，将通过修改优化的 LNP 配方。抗体偶联并在胎儿小鼠肺中测试了细胞特异性靶向。提案——作为一个跨学科项目在该部门的赞助者之间进行生物工程、佩雷尔曼医学院和费城儿童医院宾夕法尼亚大学——将允许开发一种新颖的 LNP 交付平台可应用于后续工作中，以在子宫内进行先天性肺病基因治疗。