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.

血细胞的终生生成需要由系统和系统指导的造血发育程序 传达这些细胞动态需求的局部信号如何在髓内传递。 PI 对此领域已发布和即将发布的大量信息仍知之甚少。 合作和单独的计划明确确立了聚糖在指导中的参与 造血祖细胞的命运、功能，特别是血小板生成和血小板功能。 总体假设是“细胞内在和外在聚糖介导的机制调节维持， 项目 1 将研究造血细胞的分化和功能。” 半乳糖基转移酶 β4GalT1、β1 整合素和糖胺聚糖 (GAG)/硫酸乙酰肝素蛋白聚糖 （HSPG）在稳态血小板生成和清髓应激后使用新型联合共享 项目 3 的“组学”和标准方法。β4GalT1 以前未知的调节作用 HSPG 的巨核细胞 (MK) 表达也将被研究，功能上定义的 MK 偏向。 造血干细胞将与项目2一起研究，特别是关于α2,6- 尽管缺乏生成该结构所需的 St6gal1 表达，但细胞表面仍存在唾液酸化。 2 将研究细胞外糖基化的作用，特别是由外在 ST6GAL1 介导的作用 组合“组学”方法（项目 1），如何调节造血生态位中的外源唾液酸化， 识别唾液酸化的细胞表面目标；并通过项目 3 了解新发现的 GAG 辅因子调节外源性 ST6GAL1 活性和临床骨髓增生异常综合征 (MDS)。 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具有高度异质性，但具有以下共同点： MK 发育不良以及血小板数量和功能改变，以及这些临床疾病的分析已共享 项目 3 将研究 GAG 与蛋白质的结构-功能关系。 骨髓微环境内，例如生长因子及其受体，以及糖基转移酶。 将使用多维方法探讨在促进血小板生成和细胞命运决定中的作用 识别不同的 GAG 序列 项目 3 将发现合成的 GAG 模拟物作为调节剂。 用于治疗用途的造血/血小板生成核心 A 将监督核心计划的管理。 B 将提供源自大量 RNA 样品和单细胞的 cDNA 文库的生成和测序，以及 Core C 将执行 GAG、蛋白质组学和蛋白质-GAG 相互作用的比较结构分析，以及 这三个项目紧密相连，并将使用所有核心。 该计划将发现增加血小板生成并帮助了解临床的新信息 以 MK 异常为特征的病症。