Ionizable Lipid Nanoparticles for Fetal Lung Targeting

用于胎儿肺靶向的可电离脂质纳米颗粒

• 批准号: 10464943
• 负责人: Rohan Palanki
• 金额: $ 5.18万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 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/10464943
• 关键词: 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; Epithelial Cells; Evaluation; 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; Reporter; Respiration Disorders; Respiratory Disease; Respiratory Failure; Respiratory physiology; Safety; Scheme; Slice; Source; Stains; Structure; Surface; Syndrome; System; Technology; Testing; Toxic effect; Trachea; Training; Universities; Viral Vector; Work; adenoviral-mediated; alpha 1-Antitrypsin Deficiency; base; biomaterial compatibility; cell type; clinical translation; curative treatments; cytokine; cytotoxicity; delivery vehicle; design; experience; experimental study; 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

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 仅限于姑息治疗或小儿肺移植，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 发病率和死亡率。基因编辑技术的最新进展，例如 CRISPR-Cas9。 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提供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， 运载工具将推动该领域走向临床转化。因此，该提案旨在 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。在目标 1 中，多样化。 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。