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（通过应用程序证明）：造血生长因子的投资生物效应和行动机制继续为（DYS）系统提供无关紧要的视力（DYS）系统是一个信息的重要范式。红斑基因细胞（EPC）的调节。 -32]实验室表征了epor/jak2/stat5诱导的“ erythroferrone” TNF细胞因子作为肝素抑制剂[NAT基因46：678-84]。基于修改（PTM）的LC-MS/MS/MS/MS Prot Eomics，发现有趣的EPO/EPOR/JAK2动作的新介质包括50多个以前与EPO与EPO相关的因素，该因子与EPO依赖性依赖性类别的分子适配器Hroid hroid shroid cytoskeplet蛋白，，，激酶和磷酸化和细胞周期和生存因子。（和已知的）EPO目标和换能器将被挖掘出来（与专家生物信息学家协作）。在HEPC的S，下脊椎动物中作为EPO信号传感器演变的适配器。然后，SA＃3专注于EPO作用的新下游介体，硫氧还蛋白的蛋白质（TXNIP）EPO在C端PT/PS位点的TXNIP，以及ITHENS的表达。血红蛋白细胞将通过分析EPC的生长，miRNA种群和代谢特性来确定新的EPO靶标在功能上可能与MPN和/或EPO恶化癌症结局的潜力有关。

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）

Brief report: serpin Spi2A as a novel modulator of hematopoietic progenitor cell formation.

DOI：
10.1002/stem.1778
发表时间：
2014-09
期刊：
STEM CELLS
影响因子：
5.2
作者：
Li, Lei;Byrne, Susan M.;Rainville, Nicole;Su, Su;Jachimowicz, Edward;Aucher, Anne;Davis, Daniel M.;Ashton-Rickardt, Philip G.;Wojchowski, Don M.
通讯作者：
Wojchowski, Don M.

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。

DOI：
10.1074/jbc.270.12.6619
发表时间：
1995
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Taxman,DJ;Wojchowski,DM
通讯作者：
Wojchowski,DM

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