Lysyl oxidases are novel regulators of definitive hematopoiesis

赖氨酰氧化酶是最终造血的新型调节剂

• 批准号: 10640821
• 负责人: Elizabeth Coffey
• 金额: $ 7.38万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10640821
• 关键词: Address; Animals; Aorta; Biological; Blood; Bone Marrow Transplantation; Cell Proliferation; Cell Transplantation; Cells; Clinical; Collagen; Data; Dedications; Development; Dorsal; Embryo; Embryonic Development; Endothelial Cells; Endothelium; Engraftment; Environment; Enzymes; Extracellular Matrix; Extracellular Matrix Proteins; Family; Family member; Fibronectins; Gene Transduction Agent; Generations; Genetic Transcription; Goals; Growth Factor; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Specification; Hematopoietic Stem Cell Transplantation; Hematopoietic System; Hematopoietic stem cells; Immune; In Vitro; Individual; Integrins; Investigation; Laboratories; Life; Mediating; Mediator; Mentors; Mentorship; Mesenchymal; Modeling; Modification; Molecular; Multipotent Stem Cells; Non-Malignant; PTK2 gene; Patients; Pattern; Pharmaceutical Preparations; Physical environment; Pluripotent Stem Cells; Polarization Microscopy; Population; Process; Production; Proliferating; Property; Protein-Lysine 6-Oxidase; Protocols documentation; Regulation; Research; Resource Development; Role; Saint Jude Children' s Research Hospital; Scientist; Signal Induction; Signal Transduction; Source; Specific qualifier value; Stem Cell Development; Structural Protein; Techniques; Therapeutic; Training; Vertebrates; Visualization; Zebrafish; career; career development; cell behavior; crosslink; curative treatments; delivery vehicle; differentiation protocol; directed differentiation; experience; extracellular; gene therapy; genome editing; hematopoietic stem cell expansion; hemogenic endothelium; improved; in vivo; induced pluripotent stem cell; insight; leukemia; leukemia treatment; loss of function; migration; novel; reconstitution; second harmonic; self renewing cell; stem cell biology; training opportunity; vertebrate embryos

Project Summary/Abstract Hematopoietic stem cells (HSCs) are self-renewing, transplantable cells that have the capacity to reconstitute all blood lineages for life. Consequently, HSC transplantation (HSCT) is used in treatment of leukemia and other blood disorders. In principle, induced pluripotent stem cell (iPSC)-derived HSCs could provide an unlimited source of HSCs for transplant, patient-specific in vitro studies, drug studies and as a vector for delivery of gene therapy. However, to date, it has not been possible to generate bona fide HSCs from iPSCs with high engraftment potential and multilineage reconstitution. One means of generating HSCs from iPSCs would be to reproduce the normal embryonic specification of HSCs in vitro. HSCs arise from hemogenic endothelial (HE) cells in the embryonic dorsal aorta (DA). The extracellular matrix (ECM) provides the physical environment in which HSC precursors receive signals that induce their commitment to the hematopoietic lineage. Despite the likely importance of the ECM, we do not yet understand how or whether the ECM influences HSC specification. The lysyl oxidase (Lox) family of ECM-modifying enzymes controls ECM stiffness, display of growth factor signals, and other properties by catalyzing the cross-linking of ECM proteins including collagen and fibronectin. Using the zebrafish model of hematopoietic development, I have shown that Lox activity is required for specification of the earliest HSCs. My preliminary studies indicate that at least three Lox family members are required for proper specification and maturation of the definitive hematopoietic system. In Aim 1, I will systematically examine Lox family contribution to HSC specification. In Aim 2, I will determine the mechanism of Lox regulation of HSC specification to distinguish between the most likely possibilities: Lox-induced ECM stiffness regulates HSCs directly through integrin-ECM interactions, indirectly through modulating existing growth factor signals, or both. The research and training proposed in this F32 application will take place under the guidance of my mentor, Dr. Wilson Clements and my mentorship team at St. Jude Children’s Research Hospital. The proposal describes research that will provide exceptional training opportunities and generate original and meaningful scientific findings to launch my career as an independent scientist. The aims build on my prior experience while simultaneously providing training in the field of HSC biology. I will gain expertise in genome editing and cutting- edge techniques for visualization and analysis of ECM, including second harmonic generation (SHG) and polarized light microscopy. In addition, I will take advantage of numerous career development resources.

项目摘要/摘要 造血干细胞（HSC）是具有重新构造能力的自我更新的可移植细胞 所有的血统生命。因此，HSC移植（HSCT）用于治疗白血病和其他 血液疾病。原则上，诱导多能干细胞（IPSC）衍生的HSC可以提供无限的HSC 用于移植，特定于患者的体外研究，药物研究的HSC来源，作为基因递送的载体 治疗。但是，迄今 潜力和多核心重构。从IPSC中生成HSC的一种方法是重现 HSC在体外的正常胚胎规范。 HSC是由血肿内皮（He）细胞引起的 胚胎背主动脉（DA）。细胞外基质（ECM）提供了HSC的物理环境 前体获得了影响其对造血谱系的承诺的信号。尽管有可能 ECM的重要性，我们尚不了解ECM如何或如何影响HSC规范。 ECM修饰酶的赖氨酸氧化酶（LOX）家族控制ECM刚度，显示生长因子信号， 通过催化包括胶原蛋白和纤连蛋白在内的ECM蛋白的交联，以及其他特性。使用 斑马鱼的造血发育模型，我已经表明，LOX活动是规格的 最早的HSC。我的初步研究表明，正确需要至少三个LOX家庭成员 确定造血系统的规范和成熟。 在AIM 1中，我将系统地检查LOX家庭对HSC规范的贡献。 在AIM 2中，我将确定HSC规范的LOX调节机制以区分 最可能的可能性：LOX诱导的ECM刚度直接通过整合素ECM调节HSC 通过调节现有生长因子信号的间接相互作用，或两者兼而有之。 该F32申请中提出的研究和培训将在我的导师博士的指导下进行。 威尔逊·克莱门茨（Wilson Clements）和我的圣裘德儿童研究医院（St. Judd Children's Research Hospital）的Mentalship团队。提案描述 将提供出色的培训机会的研究，并产生原始而有意义的科学 发现我作为独立科学家的职业生涯。目的是基于我以前的经验 同样，在HSC生物学领域提供培训。我将获得基因组编辑和剪裁方面的专业知识 - ECM可视化和分析的边缘技术，包括第二次谐波生成（SHG）和 偏振光显微镜。此外，我将利用众多职业发展资源。