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

• 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
• 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

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，我们利用了植物表达系统的独特优势，即 缺乏添加唾液酸的能力，无法对植物进行 asialo-rhuEPO 基因改造 - a 不含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酸 酸残基的体内功能，我们可以发现的一个例外是 rhuEPO，它具有 兼有造血和组织保护功能。当 rhuEPO 用作抗细胞凋亡剂时 剂，其造血活性可能会导致不良的副作用，导致有害的增加 红细胞团。这种活性可以通过去除唾液酸来破坏。欧洲专利局 rhuEPO 的全酶促脱唾液酸化生成的产物已被证明可以保留 EPO 的细胞和组织保护特性，无造血活性。此类EPO 衍生物可作为抗凋亡剂进行探索。然而却没有任何表情 可以直接生产asialo-rhuEPO的系统。也不太可能用于商业用途 可用 rhuEPO（仅在哺乳动物细胞中产生）来制造 asialo-rhuEPO 由于其供应有限。根据现有的知识和技术，很可能 对植物进行基因改造，生产不含唾液酸的人类糖蛋白。我们有共同 在烟草植物中表达与 ER 信号肽融合的人 GalT 基因和 EPO 基因， 并尝试廉价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asialo-rhuEPO。该项目的结果也将提供见解 深入了解植物糖基化机制和植物抗凋亡机制 产生asialo-rhuEPO。此外，蛋白质纯化方法将 糖蛋白分析方法的建立对未来的植物非常有价值 糖工程项目。

项目成果

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