Rapid egg bioreactor construction for Fabry disease therapeutics

用于法布里病治疗的快速鸡蛋生物反应器构建

基本信息

批准号：
10481061
负责人：
Qilong Ying
金额：
$ 25.95万
依托单位：
ATGC, INC.
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：
2022-08-08 至 2024-08-07
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10481061
关键词：
Affect Alpha-galactosidase Animals Biological Assay Bioreactors Blastoderm CRISPR/Cas technology Cell Culture Techniques Cell Line Cell Transplantation Cells Chicken Cells Chickens Chimera organism Chimerism Clinical Trials Code Collaborations Color Custom DNA cassette Development Drug Costs Egg White Engineering Enzymes FDA approved Fabry Disease Feathers Female Funding Future Generations Gene Targeting Gene Transfer Techniques Genetic Transcription Genotype Goals Gonadal structure Healthcare Healthcare Systems Heritability Human In Vitro Knock-in Live Birth Lysosomal Storage Diseases Mainstreaming Mammalian Cell Mammalian Oviducts Methods Mus Oryctolagus cuniculus Ovalbumin Patients Pattern Pharmaceutical Preparations Phase Price Process Production Productivity Rattus Recombinant Proteins Recombinants Secure Security Seminal fluid Serum Signal Transduction Structure of primordial sex cell System Therapeutic Tissues Transgenes Transgenic Animals Transgenic Organisms Transplantation United States National Institutes of Health Validation Wolman Disease Work chicken egg clinical development cost cost effective egg embryonic stem cell enzyme replacement therapy genome editing inhibitor male novel novel strategies pre-clinical protein expression stem cells success therapeutic enzyme therapeutic protein trait

项目摘要

Abstract Fabry disease is a lysosomal storage disease (LSD) affecting 1 in 40,000 to 117,000 live births. The current treatment is enzyme replacement therapy (ERT) with recombinant human α-galactosidase A (GalA). A major healthcare burden from this treatment is the high drug cost at $350k per patient-year. To reduce the drug cost for Fabry disease, we propose to develop a rapid approach to construct cost- effective chicken egg bioreactor for GalA production, which has the potential to produce 50 mg recombinant protein per egg for less than $10. Previously, a similar egg bioreactor was developed for another LSD, lysosomal acid lipase (LAL) deficiency. The drug sebelipase alfa for LAL deficiency has been the only FDA- approved therapeutics from transgenic chicken eggs. The traditional blastoderm transgenesis of chicken is limited by its efficiency and thousands of chickens will be screened to obtain a positive clone. Another mainstream approach with primordial germ cells (PGCs) suffers from the fact that PGCs contribute to the germline but not somatic cells, and only generate male G0 chimeras, delaying the assessment of chimerism until six months later when their semen is genotyped. Our novel approach will generate chimeric roosters and hens with both somatic and germline chimerism, which greatly accelerate transgenic chicken construction. In Phase I, we will engineer chicken cell lines with GalA expression cassette knock-in at the highly transcribed OVA locus and construct the chimeric chickens. The chimeric hens will provide eggs for rapid assessment of GalA abundance and activity. This validation will show the feasibility of this approach for the future generation of chicken with heterozygous and homozygous OVA-GalA knock-in.

抽象的法布里病是一种溶酶体储存疾病（LSD），影响40,000至117,000个活产。这当前的治疗是用重组人α-半乳糖苷酶A（GALA）的酶替代疗法（ERT）。这种治疗方法的主要医疗保健伯恩（Burnen）是每年患者$ 350K的高药物成本。为了降低Fabry病的药物成本，我们建议开发一种快速的方法来构建成本 - 有效的鸡蛋生物反应器用于盛会的生产，有可能产生50毫克重组每个鸡蛋的蛋白质少于10美元。以前，为另一个LSD开发了类似的卵生物反应器，溶酶体酸脂肪酶（LAL）缺乏症。用于LAL缺乏症的药物皮脂酶Alfa是唯一的FDA- 转基因鸡蛋的批准疗法。鸡的传统胚胚型转基因受其效率和数千只鸡的限制将被筛选以获得一个正克隆。原始生殖细胞（PGC）的另一种主流方法遭受了PGCS对种系的贡献，但不足的事实，而仅产生雄性G0 嵌合体，将嵌合的评估延迟到六个月后的精液被基因分型。我们的新颖的方法将用躯体和种系嵌合产生嵌合式公鸡和母鸡，这大大加速了转基因鸡结构。在第一阶段，我们将使用Gala表达式盒式盒式盒在高度转录OVA基因座并构造嵌合鸡。嵌合母鸡将为快速提供鸡蛋评估晚会抽象和活动。此验证将显示这种方法的可行性未来一代鸡肉，杂合和纯合子ova-gala敲击。