Developing Gene Editing Therapeutics, Biodegradable Polymeric Delivery Vehicles, and High-throughput Platforms for the Treatment of Cystic Fibrosis

开发用于治疗囊性纤维化的基因编辑疗法、可生物降解的聚合物递送载体和高通量平台

• 批准号: 10301702
• 负责人: Alexandra Sarah Annukka Piotrowski-Daspit
• 金额: $ 10.98万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10301702
• 关键词: 3-Dimensional; Amines; Base Pairing; Biocompatible Materials; Biodistribution; Biological; Biology; Biomedical Engineering; Blood Circulation; CRISPR/Cas technology; Cell Line; Cells; Chemicals; Chlorides; Clinical Trials; Communities; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; DNA; DNA Repair; Development; Development Plans; Diagnosis; Disease; Disease model; Doctor of Philosophy; Drug Delivery Systems; Encapsulated; Engineering; Esters; Family; Genes; Genetic; Genetic Diseases; Goals; Guide RNA; Half-Life; Health; Hereditary Disease; Impairment; In Vitro; Interdisciplinary Study; Knowledge; Libraries; Lung; Lung diseases; Lung infections; Mentors; Methods; Modeling; Mutation; Nanotechnology; Nonsense Codon; Oligonucleotides; Organoids; Other Genetics; Patients; Peptide Nucleic Acids; Physiological; Physiology; Polymer Chemistry; Polymers; Postdoctoral Fellow; Proteins; Public Health; RNA-Directed DNA Polymerase; Reagent; Regulator Genes; Reporter; Research; Research Personnel; Safety; Scientist; Screening procedure; Site; Structure-Activity Relationship; System; Technology; Testing; Therapeutic; Therapeutic Agents; Tissue Engineering; Tissues; Training; Translations; Treatment Efficacy; United States; Universities; Work; base; biodegradable polymer; burden of illness; career; career development; cell type; clinical translation; clinically relevant; curative treatments; cystic fibrosis patients; design; design and construction; disease-causing mutation; endonuclease; engineered nucleases; experience; experimental study; genome editing; high throughput screening; high throughput technology; improved; in vitro Model; in vivo; innovation; nanomedicine; nanoparticle; novel; nuclease; nucleic acid delivery; nucleic acid-based therapeutics; response; screening; skills; therapeutic development; therapeutic genome editing; three dimensional cell culture; three-dimensional modeling; tool; uptake

Project Summary Cystic fibrosis (CF) is a progressive genetic disorder caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. Premature stop codon mutations including W1282X are among the most severe and there are no curative treatments for patients. Genome editing agents could offer promising therapeutics applicable to all CF patients. Engineered nucleases including CRISPR/Cas9 systems that can catalyze correction of disease-causing mutation(s) have shown promise and entered clinical trials. To mitigate aberrant nuclease activity and reduce off-target effects, prime editing technology combines a catalytically impaired Cas9 endonuclease fused with an engineered reverse transcriptase programmed with a prime editing guide RNA (pegRNA) that also encodes the desired edit. As an alternative technology, triplex-forming peptide nucleic acids (PNAs) have no intrinsic nuclease activity and stimulate endogenous DNA repair with low off-target effects when bound adjacent to the target site and co-delivered with donor DNA oligonucleotides. Despite advances in gene editing technology, in vivo delivery remains a primary barrier to clinical translation. The goal of the proposed research is to develop a genome editing-based therapeutic strategy for treating the W1282X nonsense CF mutation as well as high-throughput technologies for identifying effective vehicles for in vivo therapeutic nucleic acid delivery. In Aim 1, PNA- and CRISPR/Cas9 prime editing-based gene editing reagents will be designed to correct the W1282X mutation, encapsulated into poly(amine-co-ester) (PACE) nanoparticles (NPs), and tested in vitro and in vivo. In Aim 2, novel PACE materials will be developed for in vivo delivery of nucleic acid-based therapeutics to the lungs and assessed using high-throughput in vivo platforms to determine the structure-function relationships guiding physiological fate. In Aim 3, physiologically relevant 3D culture models will be developed as high-throughput screening tools to assess delivery and efficacy of CF therapies. Overall, the proposed interdisciplinary research is highly clinically relevant, furthering the translation of promising gene editing/nucleic acid therapeutics for CF and other genetic diseases. Dr. Piotrowski-Daspit received her Ph.D. in Chemical and Biological Engineering and is currently a postdoctoral fellow in the Department of Biomedical Engineering at Yale University. Thus far, she has been developing polymeric NPs for nucleic acid delivery and high-throughput in vivo tools. The career development plan outlines a comprehensive strategy for acquiring the technical, conceptual, and professional skills required to complete the proposed studies and launch an independent research career. The proposed training would enable her to gain significant experience in therapeutic development for CF and integrate her into the CF research community. The training plan, together with her background in biomedical engineering, biomaterials and drug delivery, will place her among a select group of scientists with the skills and breadth of knowledge necessary to effectively pursue interdisciplinary work on nucleic acid delivery and editing of genetic disorders.

项目总结