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.

项目摘要先天性肺疾病，例如遗传活性剂蛋白综合征，囊性纤维化和 α-1抗胰蛋白酶缺乏，是小儿发病率和死亡率的重要来源。肺部疾病的新生儿患者的治疗选择，呼吸衰竭仅限于姑息治疗或小儿肺移植。因此，有明确的临床需求对于允许早期纠正先天性肺部疾病以减少小儿的新疗法发病率和死亡率。基因编辑技术的最新进展，例如CRISPR-CAS9 系统，解锁了纠正致病突变的潜力，从而治疗先天性来源的疾病。在子宫内执行基因编辑提供了逆转的额外好处当肺功能变为产后寿命之前，遗传异常必不可少的，并利用胎儿的正常发育特性以提高效率更正。传统上，病毒载体已用于在动物的子宫基因治疗中研究尽管这些研究令人鼓舞，但发现了替代性，可能更安全，送货车将向临床翻译前进。这是该提议的目的研究可离子脂质纳米颗粒（LNP）的潜力，这是一种有希望的非病毒递送平台，用于核酸递送到小鼠胎儿肺。胎儿肺优化脂质纳米颗粒（FLO-LNP）将通过多阶段优化方案生成。在AIM 1中，潜水员将筛选24个可离子脂质结构的库，以识别最佳的电离脂质将mRNA输送到胎儿肺部。在AIM 2中，LNP公式将通过设计实验方案，用于最小毒性和在CRISPR-CAS9系统中的最大输送方案鼠标精密切割肺切片。在AIM 3中，优化的LNP公式将通过抗体结合并测试了胎儿肺中细胞特异性靶向的测试。最终，这个提案 - 作为部门的赞助商之间的跨学科项目生物工程，佩雷尔曼医学院和费城儿童医院宾夕法尼亚大学 - 将允许开发一个新颖的LNP交付平台可以在随后的工作中应用用于先天性肺疾病的子宫基因疗法。