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EPO regulated erythropoiesis

EPO 调节红细胞生成

基本信息

批准号：
9896668
负责人：
DON Michael WOJCHOWSKI
金额：
$ 38.06万
依托单位：
UNIVERSITY OF NEW HAMPSHIRE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-25 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9896668
关键词：
Anemia Apoptosis Attenuated C-terminal Categories Cathepsins Cell Cycle Cell Membrane Permeability Cell Survival Cell membrane Cells Clinical Complex Cytoprotection Cytoskeletal Proteins Data Databases Development Dose Effector Cell Erythroblasts Erythroid Erythroid Progenitor Cells Erythropoiesis Event Genets Growth and Development function Hematopoiesis Hematopoietic Cell Growth Factors Hemoglobin Human Investigation Iron JAK2 gene Laboratories Link Lysosomes Malignant Neoplasms Mediator of activation protein Metabolic MicroRNAs Modeling Molecular Molecular Mechanisms of Action Mus Myeloproliferative disease New Agents Outcome Pathway interactions Peptides Phosphoric Monoester Hydrolases Phosphorylation Site Phosphotransferases Phosphotyrosine Population Post-Translational Protein Processing Property Protein Kinase Protein Tyrosine Phosphatase Proteomics Rattus Receptor Signaling Regulation Role Serpins Signal Transduction Site Stat5 protein Structure System TNF gene TXNIP gene Transducers Ubiquitin Vertebrates base cell growth clinically significant cytokine hepcidin insight knock-down meetings mutant novel public health relevance receptor side effect small hairpin RNA success tool transcriptome

项目摘要

DESCRIPTION (provided by applicant): Investigations of hematopoietic growth factor bio-effects and action mechanisms continue to provide important insight into (dys) regulated blood cell formation. The EPO receptor (EPOR) system is an informative and clinically significant paradigm. Recent studies applying contemporary approaches indicate major gaps in the field's understanding of EPO/EPOR/JAK2 signal transducers and their regulation of erythroid progenitor cell (EPC) formation. To illustrate, the PI has recently identified a novel EPOR/JAK2/STAT5 pathway in which an EPO-induced Spi2A serpin cytoprotects EPCs against executioner cathepsins as leached from ROS-compromised lysosomes [JEM 210:225-32]. And Dr. T. Ganz's laboratory has characterized an EPOR/JAK2/STAT5-induced "Erythroferrone" TNF cytokine as a hepcidin suppressor [Nat Genet. 46:678-84]. Via major supporting studies for this R01 renewal, we've applied post-translational modification (PTM) based LC-MS/MS proteomics to discover intriguing new mediators of EPO/EPOR/JAK2 action. These include 50+ factors not previously linked to EPO-dependent erythropoiesis within diverse functional categories of molecular adaptors, erythroid cytoskeletal proteins, kinases & phosphatases, and cell cycle & survival factors. SA#1 will extend our PTM-directed proteomic investigations in human erythroid precursor cells to include broad-based targets as modified at pY, T*PP and ubiquitin motifs, together with analyses of more select signaling nodes for S/T kinases, survival/apoptosis factors and cell cycle targets. Networks for hundreds of specifically activated PTM events for novel (and known) EPO targets and transducers will be mined (with collaborating expert bioinformaticists). SA#2 focuses on defining the functional roles and action mechanisms of three interrelated new EPO targets as upstream effectors of EPOR/JAK2 complexes. Two, C1ORF186/"RHEX" and C1ORF150, are novel molecular adaptors that have evolved as EPO signal transducers in hEPC's (but are absent among mice, rats, lower vertebrates). Third, PTPN18 is a protein tyrosine phosphatase which we demonstrate to increase JAK2 activation, decrease EPOR turnover and limit pY-RHEX formation. Approaches will include GOF, shRNA LOF, and mutant rescue studies in UT7epo cells and primary hEPCs. SA#3 then focuses on a new downstream mediator of EPO action, Thioredoxin-Interacting Protein (TXNIP). EPO modulates TXNIP at C-terminal pT/pS sites, and heightens its expression. TXNIP knockdown attenuates EPC growth, and notably accelerates primary erythroid precursor development to KIT-low, GPA-high hemoglobinizing erythroblasts. Mechanistically how TXNIP acts as a novel EPO agent will be determined by analyzing EPC growth, survival, ROS, miRNA populations and metabolic properties. Overall, studies will reveal important new mediators of EPO-dependent human erythropoiesis. Certain may be druggable with potentials to lessen EPO dosing, and limit EPO's major adverse side effects. Other new EPO targets may functionally relate to MPNs and/or to EPO's potential to worsen cancer outcomes.

描述（由申请人提供）：造血生长因子生物效应和作用机制的研究继续为（dys）调节血细胞形成提供重要见解。EPO受体（EPOR）系统是一个信息丰富且具有临床意义的范例。应用当代方法的最新研究表明，该领域对EPO/EPOR/JAK 2信号转导子及其对红系祖细胞（EPC）形成的调节的理解存在重大差距。为了说明，PI最近鉴定了一种新的EPOR/JAK 2/STAT 5途径，其中EPO诱导的Spi 2A丝氨酸蛋白酶抑制剂细胞保护EPC免受从ROS受损的溶酶体中浸出的执行组织蛋白酶的影响[JEM 210：225-32]。还有T医生Ganz的实验室已经将EPOR/JAK 2/STAT 5诱导的“Erythroferrone”TNF细胞因子表征为铁调素抑制剂[Nat Genet. 46：678-84]。通过对R 01更新的主要支持研究，我们应用了基于翻译后修饰（PTM）的LC-MS/MS蛋白质组学来发现EPO/EPOR/JAK 2作用的有趣的新介质。这些包括50多个先前未与EPO依赖性红细胞生成相关的因子，这些因子在分子衔接子、红细胞细胞骨架蛋白、激酶和磷酸酶以及细胞周期和存活因子的不同功能类别内。SA#1将扩展我们在人类红系前体细胞中的PTM指导的蛋白质组学研究，以包括在pY，T*PP和泛素基序处修饰的广泛靶点，以及对S/T激酶，存活/凋亡因子和细胞周期靶点的更多选择信号传导节点的分析。将挖掘数百个新的（和已知的）EPO靶点和转换器的特异性激活PTM事件的网络（与合作的生物信息学专家）。SA#2重点定义了作为EPOR/JAK 2复合物上游效应子的三个相互关联的新EPO靶点的功能作用和作用机制。其中两个，C1 ORF 186/“RHEX”和C1 ORF 150，是新的分子衔接子，在hEPC中进化为EPO信号转导子（但在小鼠、大鼠、低等脊椎动物中不存在）。第三，PTPN 18是一种蛋白酪氨酸磷酸酶，我们证明它可以增加JAK 2的激活，减少EPOR的周转，并限制pY-RHEX的形成。方法将包括在UT 7 epo细胞和原代hEPCs中的GOF、shRNA LOF和突变体拯救研究。SA#3然后关注EPO作用的新下游介质，硫氧还蛋白相互作用蛋白（TXNIP）。EPO在C-末端pT/pS位点调节TXNIP，并提高其表达。TXNIP敲低减弱EPC生长，并显著加速初级红细胞前体发育为KIT低、GPA高的血红蛋白化成红细胞。TXNIP作为一种新型EPO药物的机制将通过分析EPC生长、存活、ROS、miRNA群体和代谢特性来确定。总体而言，研究将揭示EPO依赖性人红细胞生成的重要新介质。某些可能是可药用的，有可能减少EPO的剂量，并限制EPO的主要不良副作用。其他新的EPO靶点可能在功能上与MPN和/或EPO恶化癌症结果的潜力有关。

项目成果

期刊论文数量（48）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A dimeric peptide with erythropoiesis-stimulating activity uniquely affects erythropoietin receptor ligation and cell surface expression.

具有红细胞生成刺激活性的二聚肽独特地影响红细胞生成素受体连接和细胞表面表达。

DOI：
10.1016/j.exphem.2016.04.015
发表时间：
2016
期刊：
Experimental hematology
影响因子：
2.6
作者：
Verma,Rakesh;Green,JenniferM;Schatz,PeterJ;Wojchowski,DonM
通讯作者：
Wojchowski,DonM

Phosphorylatable and epitope-tagged human erythropoietins: utility and purification of native baculovirus-derived forms.

可磷酸化和表位标记的人类促红细胞生成素：天然杆状病毒衍生形式的用途和纯化。

DOI：
10.1016/1046-5928(92)90063-3
发表时间：
1992
期刊：
Protein expression and purification
影响因子：
1.6
作者：
Quelle,DE;Lynch,KJ;Burkert-Smith,RE;Weiss,S;Whitford,W;Wojchowski,DM
通讯作者：
Wojchowski,DM

Dynamic ligand modulation of EPO receptor pools, and dysregulation by polycythemia-associated EPOR alleles.

EPO 受体库的动态配体调节以及红细胞增多症相关 EPOR 等位基因的失调。

DOI：
10.1371/journal.pone.0029064
发表时间：
2012
期刊：
PloS one
影响因子：
3.7
作者：
Singh S;Verma R;Pradeep A;Leu K;Mortensen RB;Young PR;Oyasu M;Schatz PJ;Green JM;Wojchowski DM
通讯作者：
Wojchowski DM

Comparative analysis of the locus control region of the rabbit beta-like gene cluster: HS3 increases transient expression of an embryonic epsilon-globin gene.

兔β样基因簇基因座控制区的比较分析：HS3增加胚胎ε-珠蛋白基因的瞬时表达。

DOI：
10.1093/nar/21.5.1265
发表时间：
1993
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Hardison,R;Xu,J;Jackson,J;Mansberger,J;Selifonova,O;Grotch,B;Biesecker,J;Petrykowska,H;Miller,W
通讯作者：
Miller,W

Erythropoietin-induced transcription at the murine beta maj-globin promoter. A central role for GATA-1.

促红细胞生成素在鼠β大球蛋白启动子处诱导转录。