Molecular and Clinical Glycobiology of the Bone Marrow Environment

骨髓环境的分子和临床糖生物学

• 批准号: 10545004
• 负责人: Karin Maria Hoffmeister
• 金额: $ 245.99万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10545004
• 关键词: Abnormal megakaryocyte; Accidents; Address; Affect; Affinity Chromatography; Automobile Driving; Biology; Blood; Blood Cell Count; Blood Cells; Blood Platelets; Bone Marrow; Burn injury; Cell surface; Cells; Cellular Structures; Clinical; Cues; Data; Development; Dimensions; Disease; Disease Progression; Dysmyelopoietic Syndromes; Dysplastic Megakaryocyte; Environment; Functional disorder; Galactosyltransferases; Gene Expression; Generations; Glycobiology; Glycosaminoglycans; Goals; Growth Factor; Health; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Heparan Sulfate Proteoglycan; Human; Iatrogenesis; Individual; Integrins; Joints; Knowledge; Libraries; Life; Maintenance; Marrow; Mediating; Megakaryocytes; Molecular; Mus; Myelodysplastic/Myeloproliferative Disease; Myeloproliferative disease; Platelet Count measurement; Polysaccharides; Population; Process; Production; Proteins; Proteomics; Publishing; RNA; Radio; Regulation; Role; Sampling; Sialyltransferases; Signal Transduction; Source; Specimen; Stress; Structure; Structure-Activity Relationship; Sulfate; Therapeutic; Therapeutic Uses; Thrombocytopenia; Thrombopoiesis; Time; cDNA Library; cofactor; comparative; extracellular; glycosylation; glycosyltransferase; insight; mimetics; mouse model; new therapeutic target; novel; platelet function; pre-clinical; programs; protein expression; receptor; restoration; sialylation; stem cells; sugar

The life-long production of blood cells requires hematopoietic developmental programs guided by systemic and local signals to convey the dynamic need for these cells. How these signals are delivered within the intra-medullar niches remain poorly understood. Extensive published and to-be-published information by the PIs of this Program, collaborative and individually, clearly established the involvement of glycans in the guidance of hematopoietic progenitor cell fate, function, and especially in thrombopoiesis and platelet functionality. The overarching hypothesis is “cell-intrinsic and extrinsic glycan-mediated mechanisms regulate maintenance, differentiation, and function of hematopoietic cells.” Project 1 will investigate the roles of the galactosyltransferase β4GalT1, β1 integrin, and glycosaminoglycans (GAGs)/heparan sulfate proteoglycans (HSPGs) in thrombopoiesis at steady-state and following myeloablative stress using novel combined shared “omics” and standard approaches with Project 3. A previously unknown role of β4GalT1 to regulate megakaryocyte (MK) expression of HSPGs will also be investigated. A functionally defined MK-biased hematopoietic stem cell will be investigated together with Project 2, especially with respect to the heavily α2,6- sialylated cell surface despite the absence of St6gal1 expression necessary to generate this structure. Project 2 will investigate the role of extracellular glycosylation, especially that mediated by extrinsic ST6GAL1 using the combined “omics” approach (Project 1), how extrinsic sialylation in the hematopoietic niche is regulated, identifying the cell surface targets of sialylation; and with Project 3, understanding how the newly discovered GAG cofactor modulates extrinsic ST6GAL1 activity. Clinical Myelodysplastic Syndromes (MDS) and Myeloproliferative Neoplasms (MPN) and preclinical mouse models will be used in a first-time assessment into the glycobiology of these marrow diseases of highly heterogenous presentations but with the commonality of dysplastic MKs and altered platelet numbers and function, and analysis of these clinical diseases is shared across all three Projects. Project 3 will investigate the structure–function relationships of GAGs with proteins within the marrow microenvironment, such as growth factors and their receptors, and glycosyltransferases. Their roles in promoting thrombopoiesis and cell fate decisions will be interrogated using a multi-dimensional approach to identifying distinct GAG sequences. Project 3 will discover synthetic GAG mimetics as modulators of hematopoiesis/thrombopoiesis for therapeutic use. Core A will oversee the administration of the program, Core B will provide generation and sequencing of cDNA libraries derived from bulk RNA samples and single cells, and Core C will perform comparative structural analysis of GAGs, proteomics and protein-GAG interactions, and quantitative proteomics of protein expression. The three projects are intimately intertwined and will use all Cores. This program will uncover novel information to increase platelet production and help understand clinical conditions characterized by MK abnormalities.

终身产生的血细胞需要由全身和 局部信号传达这些细胞的动态需求。这些信号是如何在Medullar内部传递的 壁ni的理解仍然很少。 PIS的广泛发布和发布的信息 计划合作和单独的计划清楚地确定了聚糖在指导下的参与 造血祖细胞命运，功能，尤其是在血小板和血小板功能中。 总体假设是“细胞中性和外部聚糖介导的机制，可调节维持， 造血细胞的分化和功能。”项目1将研究 半乳糖基转移酶β4GALT1，β1整合素和糖胺聚糖（GAGS）/硫酸乙酰肝素蛋白聚糖 （HSPGS）在稳态下的血小板中，并使用新型组合共享 项目3的“ OMICS”和标准方法。 还将研究HSPG的巨核细胞（MK）表达。功能定义的MK偏置 造血干细胞将与项目2一起研究，尤其是关于严重的α2,6-- 尽管没有ST6GAL1表达来产生这种结构，但硫化细胞表面尽管没有ST6GAL1表达。项目 2将研究细胞外糖基化的作用，尤其是使用外部ST6GAL1介导的作用 联合的“ OMICS”方法（项目1），如何调节造血生态裂的外在溶解度 识别乙型苷的细胞表面靶标；并了解项目3，了解新发现的方式 GAG辅因子调节外部ST6GAL1活性。临床骨髓发育异常综合征（MDS）和 骨髓增生性肿瘤（MPN）和临床前小鼠模型将用于首次评估 这些骨髓疾病的糖生物学是高度异质表现的，但具有 发育不良的MK和血小板数量和功能变化，分析这些临床疾病 在所有三个项目中。项目3将研究插孔与蛋白质的结构 - 功能关系 在骨髓微环境中，例如生长因子及其受体以及糖基转移酶。他们的 在促进血小板和细胞脂肪决策中的作用将使用多维方法进行询问 识别不同的插孔序列。项目3将发现合成的插科打模拟物作为调节剂 用于治疗的造血/血小板。核心A将监督该计划的管理，核心 b将提供源自大量RNA样品和单个细胞的cDNA文库的生成和测序，以及 核心C将对插科打a，蛋白质组学和蛋白质 - 蛋白质相互作用的比较结构分析以及 蛋白质表达的定量蛋白质组学。这三个项目紧密地交织在一起，将使用所有核心。 该程序将发现新的信息以增加血小板的产生并帮助了解临床 Mk异常的特征条件。