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Plant-produced Asialo-erythropoietin and its Antiapoptotic Functions

植物性促红细胞生成素及其抗细胞凋亡功能

基本信息

批准号：
7693150
负责人：
Jiahua Xie
金额：
$ 10.44万
依托单位：
NORTH CAROLINA CENTRAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7693150
关键词：
Academia Adverse effects Advocate Amino Acids Animal Model Applications Grants Appointment Area Biological Process Biological Products Biomanufacturing Biomass Biomedical Engineering Bioreactors Biotechnology C-terminal Cardiac Myocytes Cardiotonic Agents Cell Culture Techniques Cell Death Cell Line Cells Codon Nucleotides Collaborations Communities Complex Data Development Discipline Educational process of instructing Erythrocytes Erythropoietin Faculty Foundations Fucose Funding Future Galactose Galactosyltransferases Gene Expression Genes Genetic Genetically Modified Plants Glycoproteins Goals Grant Harvest Hematopoietic Hormones Human Immunoblotting Industry Institutes Institution Kanamycin Resistance Knowledge Laboratories Left Link Mammalian Cell Mammals Measures Medical Methodology Methods Minority Mission Modification Molecular Molecular Biology Mus Neurons North Carolina Oxygen PC12 Cells Pattern Peptide Signal Sequences Pharmacologic Substance Plant Leaves Plants Play Polysaccharides Process Production Productivity Property Protein Chemistry Proteins Protocols documentation Publications Publishing Research Research Activity Research Institute Research Personnel Research Support Risk Role Science Seeds Sialic Acids Solid Structure Students System Techniques Technology Testing Tissues Tobacco Tobacco use Training Transgenic Organisms Transgenic Plants Universities Vision Work Xylose base career cost forest glycosylation hematopoietic tissue improved in vivo insight interest large scale production novel pathogen prevent professor protein purification public health relevance receptor recombinant human erythropoietin scale up segregation sugar symposium systems research vector

项目摘要

DESCRIPTION (provided by applicant): In this project, we are taking the distinct advantage of plant expression systems, which is the lack of the capacity to add sialic acids, to genetically modify plants for asialo-rhuEPO - a special glycoprotein without sialic acids. Although nearly all human glycoproteins require sialic acid residues for their in vivo functions, the one exception we could find is rhuEPO, which has both hematopoietic and tissue protective functions. When rhuEPO is used as an antiapoptotic agent, its hematopoietic activity can cause bad side effects by leading to harmful increases in red blood cell mass. This activity can be disrupted by removing sialic acids. Asialo-rhuEPO generated by total enzymatic desialylation of rhuEPO has been demonstrated to retain the cell- and tissue-protective properties of EPO without hematopoietic activity. This type of EPO derivative can be explored as the antiapoptotic agent. However, there is no expression system that can produce asialo-rhuEPO directly. It is also unlikely to use commercially available rhuEPO, which is produced only in mammalian cells, to make asialo-rhuEPO because of its limited supply. Based on available knowledge and techniques, it is likely to genetically modify plants to produce human glycoproteins without sialic acids. We have co- expressed human GalT gene and EPO gene fusing with ER-signal peptides in tobacco plants, and try to inexpensively produce asialo-rhuEPO with the potential of large scale production. We expect plant-produced asialo-rhuEPO will have similar cytoprotective function as the enzymatically produced asialo-rhuEPO. The results of this project will also provide insights into the plant glycosylation mechanism and the antiapoptotic mechanism of the plant- produced asialo-rhuEPO. In addition, the protein purification methodologies that will be established and the glycoprotein analysis methods will be very valuable for future plant glycoengineering projects. PUBLIC HEALTH RELEVANCE: We aim to genetically modify plants for production of asialo-rhuEPO - a special human protein. This type of biopharmaceutical protein can be explored as the neurotherapeutic and cardioprotective agent. Using a plant expression system, we expect to produce an inexpensive and functional asialo-rhuEPO to prevent cell death with the potential of large scale production.

描述（由申请人提供）：在这个项目中，我们利用植物表达系统缺乏添加唾液酸的能力的独特优势，对植物进行基因改造，以获得asialo-rhuEPO——一种不含唾液酸的特殊糖蛋白。尽管几乎所有人类糖蛋白的体内功能都需要唾液酸残基，但我们发现的一个例外是rhuEPO，它同时具有造血和组织保护功能。当 rhuEPO 用作抗细胞凋亡剂时，其造血活性会导致红细胞量有害增加，从而产生不良副作用。这种活性可以通过去除唾液酸来破坏。通过 rhuEPO 的全酶促去唾液酸化产生的 Asialo-rhuEPO 已被证明保留了 EPO 的细胞和组织保护特性，而没有造血活性。此类EPO衍生物可作为抗凋亡剂进行探索。然而，目前还没有可以直接产生asialo-rhuEPO的表达系统。由于供应有限，也不太可能使用市售的 rhuEPO（仅在哺乳动物细胞中产生）来制造 asialo-rhuEPO。根据现有的知识和技术，有可能对植物进行基因改造以生产不含唾液酸的人类糖蛋白。我们在烟草植物中共表达了与ER信号肽融合的人GalT基因和EPO基因，并尝试廉价地生产具有大规模生产潜力的asialo-rhuEPO。我们预计植物产生的 asialo-rhuEPO 将具有与酶法产生的 asialo-rhuEPO 类似的细胞保护功能。该项目的结果还将为植物糖基化机制和植物产生的asialo-rhuEPO的抗凋亡机制提供见解。此外，将建立的蛋白质纯化方法和糖蛋白分析方法对于未来的植物糖工程项目非常有价值。公共健康相关性：我们的目标是对植物进行基因改造，以生产 asialo-rhuEPO（一种特殊的人类蛋白质）。这种类型的生物制药蛋白可以作为神经治疗和心脏保护剂进行探索。使用植物表达系统，我们期望生产一种廉价且功能性的asialo-rhuEPO，以防止细胞死亡，并具有大规模生产的潜力。