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：
8326101
负责人：
Jiahua Xie
金额：
$ 10.4万
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 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8326101
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 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 receptor recombinant human erythropoietin scale up segregation sugar symposium systems research vector

项目摘要

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.

在这个项目中，我们正在利用植物表达系统的明显优势，这是缺乏添加唾液酸的能力，可以使植物改变植物的asialo -rhuepo -a 没有唾液酸的特殊糖蛋白。尽管几乎所有人类糖蛋白都需要唾液其体内功能的酸残留物，我们可以找到的一个例外是Rhuepo，它具有造血和组织保护功能。当rhuepo用作抗凋亡特工，其造血活性会通过导致有害增加而导致不良副作用红细胞质量。可以通过去除唾液酸来破坏这种活性。 asialo-rhuepo 已证明由Rhuepo的总酶脱酰化产生没有造血活性的EPO的细胞和组织保护特性。这种类型的EPO 衍生物可以作为抗凋亡剂探索。但是，没有表达可以直接产生Asialo-Rhuepo的系统。它也不太可能在商业上使用仅在哺乳动物细胞中生产的Rhuepo可用来制作asialo-rhuepo 由于其供应有限。基于可用的知识和技术，它很可能基因修饰植物以产生无唾液酸的人类糖蛋白。我们有共同在烟草植物中与ER信号肽融合的人类Galt基因和EPO基因融合，并尝试廉价地生产具有大规模生产的潜力的Asialo-Rhuepo。我们预计植物生产的asialo-rhuepo将具有类似的细胞保护功能酶促产生的asialo-rhuepo。该项目的结果还将提供见解进入植物糖基化机制和植物的抗凋亡机制生产Asialo-Rhuepo。另外，蛋白质纯化方法将是建立和糖蛋白分析方法对于将来的植物非常有价值糖工程项目。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Two-step purification procedure for recombinant human asialoerythropoietin expressed in transgenic plants.

DOI：
10.1016/j.ijbiomac.2014.10.033
发表时间：
2015-01
期刊：
International journal of biological macromolecules
影响因子：
8.2
作者：
Kittur FS;Arthur E;Nguyen M;Hung CY;Sane DC;Xie J
通讯作者：
Xie J

Differential expression of a novel gene EaF82a in green and yellow sectors of variegated Epipremnum aureum leaves is related to uneven distribution of auxin.

一种新基因 EaF82a 在杂色 Epipremnum aureum 叶子的绿色和黄色部分中的差异表达与生长素的不均匀分布有关。

DOI：
10.1111/ppl.12219
发表时间：
2014
期刊：
Physiologia plantarum
影响因子：
6.4
作者：
Hung,Chiu-Yueh;Umstead,MakendraL;Chen,Jianjun;Holliday,BronwynM;Kittur,FarooqahmedS;Henny,RichardJ;Burkey,KentO;Xie,Jiahua
通讯作者：
Xie,Jiahua