Erythropoietin in Non-Erythroid Cells: Function and Regulation

非红细胞中的促红细胞生成素：功能和调节

• 批准号: 8195889
• 负责人: JOSEF T PRCHAL
• 金额: --
• 依托单位: VA SALT LAKE CITY HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-10-01 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8195889
• 关键词: Address; Afghanistan; Aging; Alleles; Anemia; Animal Model; Atherosclerosis; Binding; Bioinformatics; Biological Assay; Blood; Blood Circulation; Blood Vessels; Brain; Breast; Cancer Patient; Cardiovascular Diseases; Cardiovascular system; Cell physiology; Cells; Cerebrovascular Disorders; ChIP-seq; Clinical; Clinical Research; Cognitive; Congenital Disorders; Crossbreeding; Cytomegalovirus; Data; Disease; EPO gene; EPOR gene; Elderly; Elements; Embryo; Endothelium; Erythrocytes; Erythroid; Erythroid Cells; Erythropoietin; Exhibits; Foundations; Future; Gene Expression Regulation; Gene Targeting; Generations; Genes; Genetic Transcription; Genotype; Germ Cells; Goals; Grant Review; Health; Healthcare; Hematopoietic; Hematopoietic stem cells; Human; Hypertension; Hypoxia; Hypoxia Inducible Factor; Hypoxia-Responsive Elements; Individual; Inherited; Institutes; Institution; Institutional Review Boards; Intervention; Introns; Investigation; Iraq; Ischemia; Kidney Failure; Knock-in Mouse; Knockout Mice; Knowledge; Malignant - descriptor; Malignant Neoplasms; Membrane; Methods; Modeling; Molecular; Mus; Mutation; Nervous system structure; Neurologic; Normal tissue morphology; Oligonucleotides; Oocytes; Ovary; Pathology; Patients; Persian Gulf; Pharmacologic Substance; Play; Polycythemia; Population; Positioning Attribute; Prevention; Prevention strategy; Principal Investigator; Production; Protocols documentation; Receptor Gene; Regulation; Relative (related person); Renal carcinoma; Reporter; Reporting; Research; Role; Signal Transduction; Site; Specificity; Stress; Stroke; Technology; Testing; Testis; Therapeutic; Tissues; Transcript; Transgenic Mice; Traumatic Brain Injury; Universities; Utah; Variant; Veterans; abstracting; bHLH-PAS factor HLF; base; chromatin immunoprecipitation; clinical effect; clinically significant; cognitive function; erythroid differentiation; gain of function; genome-wide; high risk; hypoxia inducible factor 1; improved; instrument; interest; mouse model; novel; progenitor; programs; promoter; receptor; recombinant human erythropoietin; response; transcription factor

PROJECT SUMMARY/ABSTRACT Erythropoietin (Epo) is the principle regulator of red cell production, as it controls the survival, proliferation, and differentiation of erythroid progenitors. Recombinant human Epo has been widely used to correct the anemia of renal failure as well as other types of anemias. However, diverse clinical effects of Epo in nonerythroid tissues have been reported and transcripts of EPO and/or its receptor (EPOR) genes are found in brain, endothelium, hematopoietic stem cells, breast, ovary, testes, and other tissues. Reported beneficial effects of Epo include its neuroprotective role in hypoxic stress observed in the ischemic brains of stroke patients, while other studies demonstrated that Epo administration can also be detrimental. Levels of Epo and EpoR, regulated by HIF-1 and/or HIF-2, are reported to be associated with decreased survival of cancer patients and increased cancer aggressiveness in several clinical studies, and some clinical studies of Epo therapy in cancer patient suggested that Epo administration can be detrimental. Further, EPO and EPOR transcripts have been found in some normal tissues as well as in many cancers. Due to the embryonic lethality of Epo or EpoR null mice and the absence of a proper animal model, the study of Epo function in nonerythroid cells has been limited. Previously, we generated two viable knock-in mouse models with reduced Epo signaling (these mice are anemic) and with augmented Epo signaling (these mice are polycythemic). These mice are the foundation for studying Epo function in nonerythroid tissue. We have already generated erythroid-specific Cre-expressing mice. These mice will be used to restore Epo signaling only in blood, thus rescuing the transgenic mice from anemia. These mice will be used for the generation of a gain-of-function EpoR mouse in tissues of interest such as brain, by crossbreeding with brain-specific cre mice. Hypoxia-inducible factors (HIF) are the key transcription factors that are induced by hypoxia (discovered from studies of EPO gene regulation) and that are elevated in cancer. HIFs are heterodimers that consist of an ¿- subunit (3 isotypes; HIF-1¿, HIF-2¿, HIF-3¿) and a common b-subunit. HIF-1¿ and HIF-2¿ have significant homology, and while they may share regulation of some of their target genes, in some instances their gene regulation is tissue- and gene-specific. The molecular basis of HIF regulation is under intense investigation. While Epo was the first identified gene with a defined oligonucleotide sequence for HIF binding that was designated as Hypoxia Responsive Element (HRE), the EpoR gene does not have a conserved HRE. We have studied the mechanism of HIF regulation of Epo transcription using chromatin immunoprecipitation (ChIP) and have found multiple HREs of the Epo gene. Our data suggest that these HREs are differentially utilized by HIF- 1 and HIF-2 in a tissue-specific manner. Based on this evidence, we propose to study HRE in selected tissues under hypoxic and normoxic conditions using a genome-wide Chip-Seq instrument purchased by our institution. IMPACT ON VETERANS HEALTHCARE: Aging veterans, as well as newly returning veterans from Persian Gulf region, suffer from cognitive, vascular, and malignant disorders. The data obtained from this project will be used to characterize augmented Epo/EpoR signaling reported in some human malignancies and may serve as potential therapeutic cancer targets, while in other situations (i.e., strokes) the therapeutic augmentation of Epo/EpoR in selected tissues may be beneficial. Our ongoing and future planned studies should clarify the effect of Epo on atherosclerosis, hypertension, cognitive function, and prevention of ischemia-induced tissue damage.

项目摘要/摘要 红细胞生成素（EPO）是红细胞产生的主要调节剂，因为它控制了生存，增殖， 和红细胞祖细胞的分化。重组人EPO已被广泛用于纠正 肾衰竭以及其他类型的贫血的贫血。但是，EPO在 已经报道了非五个组织，并且在EPO和/或其接收器（EPOR）基因的转录本中发现 脑，内皮，造血干细胞，乳房，卵巢，睾丸和其他组织。报道了有益的效果 EPO包括其在中风患者缺血大脑中观察到的神经保护作用， 尽管其他研究表明也可以确定EPO给药。 EPO和EPOR的水平， 据报道，受HIF-1和/或HIF-2调节，据报道与癌症患者的存活降低有关 在几项临床研究中提高了癌症的侵略性，以及癌症中EPO治疗的一些临床研究 患者建议可以确定EPO给药。此外，EPO和Epor成绩单已经 在某些正常组织以及许多癌症中发现。由于EPO或Epor Null的胚胎致死性 小鼠和缺乏适当的动物模型，肾脏刺激细胞中的EPO功能的研究一直是 有限的。以前，我们生成了两个可行的敲入小鼠模型，其EPO信号传导降低（这些小鼠 是贫血），具有增强的EPO信号传导（这些小鼠是多余的）。这些老鼠是基础 用于研究肾脏组织中的EPO功能。我们已经产生了红骨特异性的表达 老鼠。这些小鼠将仅在血液中恢复EPO信号，从而从 贫血。这些小鼠将用于生成感兴趣的组织中功能的Epor小鼠 作为大脑，通过与脑特异性CRE小鼠杂交。 低氧诱导因素（HIF）是由缺氧诱导的关键转录因子（从中发现 EPO基因调节的研究）和癌症中升高的研究。 HIF是由�组成的异二聚体 亚基（3个同型； HIF-1，HIF-2？，HIF-3？）和一个常见的B-subunit。 HIF-1和HIF-2？很重要 同源性，虽然它们可能对某些靶基因进行调节，但在某些情况下是其基因 调节是组织和基因特异性的。 HIF调节的分子基础正在进行严格的研究。 虽然EPO是第一个具有定义的HIF结合寡核苷酸序列的基因 Epor基因被指定为缺氧反应元件（HRE），没有保守的HRE。我们有 使用染色质免疫沉淀（CHIP）和 已经找到了多个EPO基因的HRE。我们的数据表明，这些HRE被HIF-使用不同 1和HIF-2以组织特异性方式。基于此证据，我们建议研究选定组织中的HRE 在低氧和常规条件下，使用我们机构购买的全基因组芯片式仪器。 对退伍军人医疗保健的影响：老年退伍军人以及波斯的新退伍军人 海湾地区患有认知，血管和恶性疾病。从该项目获得的数据将 用于表征某些人类恶性肿瘤中报道的增强的EPO/EPOR信号传导，可以用作 潜在的治疗癌症靶点，而在其他情况下（即中风）的治疗性增强 选定组织中的EPO/EPOR可能是有益的。我们正在进行的和未来的计划研究应该澄清 EPO对动脉粥样硬化，高血压，认知功能以及缺血诱导的组织的影响 损害。