Generation of High Impact Resources for Erythropoietin Receptor Research

为促红细胞生成素受体研究生成高影响力资源

• 批准号: 8071128
• 负责人: Orson W Moe
• 金额: $ 17.61万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8071128
• 关键词: Address; Agonist; Animal Model; Apoptosis; Area; Binding; Binding Proteins; Biology; Cell membrane; Cell physiology; Clinical; Collaborations; Communities; Cytoplasmic Protein; DNA; Data; Discipline; EPOR gene; Endocrine; Erythropoiesis; Erythropoietin; Erythropoietin Receptor; Extracellular Domain; Funding; Future; Generations; Genetic Transcription; Health Benefit; Immune Sera; Ischemia; Knowledge; Laboratories; Lead; Length; Libraries; Mediating; Membrane Proteins; Messenger RNA; Methods; Modeling; Nature; Organ; Organogenesis; Peptoids; Principal Investigator; Process; Protein Binding; Proteins; Public Domains; Public Health; Reagent; Receptor Signaling; Regulation; Research; Research Personnel; Resource Sharing; Resources; Signal Pathway; Signal Transduction; Testing; Therapeutic; Therapeutic Agents; Tissues; Transcend; Transcript; Translating; Uncertainty; United States National Institutes of Health; Validation; Work; Wound Healing; angiogenesis; autocrine; carcinogenesis; drug discovery; follow-up; high risk; in vivo; novel; novel strategies; novel therapeutics; paracrine; polypeptide; protein expression; protein protein interaction; public health relevance; receptor; receptor binding; receptor expression; repaired; small molecule; therapeutic development; tool

DESCRIPTION (provided by applicant): Erythropoietin (EPO) signals via its receptor (EPOR) to exert widespread endocrine (erythropoiesis) and paracrine-autocrine (proliferation, apoptosis, angiogenesis, organogenesis, cyto-protection, repair, and carcinogenesis) actions. Progress in EPOR research has been hampered by distinct gaps in knowledge and tools: A) Uncertainty regarding the existence of "antisense EPOR polypeptides". B) Paucity of molecules (agonists or antagonists) binding to EPOR other than EPO itself. C) Incomplete knowledge of binding partners of EPOR through protein-protein interaction. We propose to propel the EPOR field forward by employing focused, short-term, intense efforts that close these gaps using conventional and novel approaches to generate high impact reagents and fundamental data that will be applicable to multiple disciplines. Specific Aims are to: 1) Follow up on the discovery of natural complementary antisense EPOR (asEPOR) transcripts that synergistically enhance EPOR protein expression induced by the sense EPOR mRNA, by determining whether asEPOR transcripts are translated into "antisense polypeptides" in vivo. 2) Identify peptoids that bind to, activate or inactivate EPOR using existing recombinatorial peptoid libraries; peptoid agonists and antagonists can serve as lead compounds for discovery of therapeutic agents. 3) Identify EPOR- binding membrane and cytoplasmic proteins using a novel approach where full length EPOR serves as bait. Identifying EPOR binding partners will open new horizons for manipulating EPOR signaling pathways. These Aims involve 3 investigators and 2 collaborating laboratories with different expertise and resources. With intense work in 2 years, we aim to deliver new reagents and knowledge, and generate resources for the entire EPOR research community. Reagents include 1) If in vivo asEPOR polypeptide is proven, specific antisera will be available. 2) EPOR-modifying peptoids to spark therapeutic drug discovery and for manipulating EPOR signaling in animal models. Knowledge includes 1) Proof or disproof of in vivo asEPOR polypeptides. Proof of asEPOR polypeptides will jumpstart a novel area of research, while disproof will focus future work on regulatory mechanisms of asEPOR transcripts. 2) A complete list of transmembrane and cytoplasmic EPOR binding proteins will be identified and validated to facilitate the study of EPOR signaling. Our approach is discovery in nature rather than hypothesis testing. PUBLIC HEALTH RELEVANCE: Erythropoietin receptor biology impacts basic biomedical as well as clinical disciplines. There is high potential for clinical therapeutic development; hence unequivocal public health benefit to having these results in the public domain. Knowledge and reagents from this project integrate immediately with other NIH funded research.

说明(申请人提供)：促红细胞生成素(EPO)通过其受体(EPOR)发出信号，发挥广泛的内分泌(红细胞生成)和旁分泌-自分泌(增殖、凋亡、血管生成、器官生成、细胞保护、修复和致癌)作用。EPOR研究的进展受到知识和工具方面的明显差距的阻碍：a)关于“反义EPOR多肽”存在的不确定性。B)缺乏与EPOR结合的分子(激动剂或拮抗剂)，而不是EPO本身。C)对通过蛋白质-蛋白质相互作用的EPOR结合伙伴的不完全了解。我们建议通过采用集中的、短期的、密集的努力来推动EPOR领域向前发展，这些努力利用传统和新的方法来缩小这些差距，以产生适用于多个学科的高影响试剂和基础数据。具体目的是：1)研究发现天然互补反义EPOR(AsEPOR)转录本，通过确定其在体内是否被翻译成反义多肽，协同增强正义EPOR mRNA诱导的EPOR蛋白表达。2)利用现有的重组类肽文库鉴定与EPOR结合、激活或失活的类肽；类肽激动剂和拮抗剂可以作为发现治疗药物的先导化合物。3)采用以全长EPOR为诱饵的新方法鉴定EPOR结合膜和细胞质蛋白。识别EPOR结合伙伴将为操纵EPOR信号通路开辟新的视野。这些目标涉及3名调查人员和2个拥有不同专门知识和资源的合作实验室。通过两年的紧张工作，我们的目标是提供新的试剂和知识，并为整个EPOR研究社区产生资源。试剂包括：1)如果EPOR多肽在体内被证实，将有特异的抗血清可用。2)在动物模型中修饰EPOR多肽以激发治疗药物的发现和操纵EPOR信号。知识包括1)体内作为EPOR多肽的证明或反证。对asEPOR多肽的证明将启动一个新的研究领域，而反证将把未来的工作重点放在asEPOR转录本的调控机制上。2)将鉴定和验证跨膜和细胞质EPOR结合蛋白的完整清单，以促进EPOR信号转导的研究。我们的方法是本质上的发现，而不是假设检验。 公共卫生相关性：促红细胞生成素受体生物学影响基础生物医学和临床学科。临床治疗开发具有很高的潜力；因此，将这些结果公之于众，无疑有利于公共卫生。来自该项目的知识和试剂立即与NIH资助的其他研究相结合。