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.

项目概要/摘要 造血干细胞 (HSC) 是自我更新、可移植的细胞，具有重建能力 所有血统均经过终生测试，造血干细胞移植（HSCT）用于治疗白血病和其他疾病。 原则上，诱导多能干细胞 (iPSC) 衍生的 HSC 可以提供无限的治疗。 用于移植、患者特异性体外研究、药物研究以及作为基因传递载体的 HSC 来源 然而，迄今为止，还不可能从具有高植入率的 iPSC 中产生真正的 HSC。 从 iPSC 生成 HSC 的一种方法是复制 体外 HSC 的正常胚胎规格源自造血内皮 (HE) 细胞。 胚胎背主动脉 (DA) 细胞外基质 (ECM) 为 HSC 提供了物理环境。 尽管有可能，前体细胞接收到诱导其致力于造血谱系的信号。 由于 ECM 的重要性，我们尚不清楚 ECM 如何或是否影响 HSC 规范。 ECM 修饰酶的赖氨酰氧化酶 (Lox) 家族控制 ECM 硬度、生长因子信号的显示、 通过催化 ECM 蛋白（包括胶原蛋白和纤连蛋白）的交联来实现其他特性。 在斑马鱼造血发育模型中，我已经证明 Lox 活性是规范 我的初步研究表明，至少需要三名 Lox 家族成员才能获得适当的 HSC。 最终造血系统的规范和成熟。 在目标 1 中，我将系统地研究 Lox 家族对 HSC 规范的贡献。 在目标 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硬度通过整合素-ECM直接调节HSC 相互作用，间接通过调节现有的生长因子信号，或两者兼而有之。 此 F32 申请中提出的研究和培训将在我的导师 Dr. 威尔逊·克莱门茨和我在圣裘德儿童研究医院的指导团队描述了该提案。 研究将提供特殊的培训机会并产生原创且有意义的科学 这些发现以我之前的经验为基础，开启了我作为一名独立科学家的职业生涯。 同时提供 HSC 生物学领域的培训，我将获得基因组编辑和切割方面的专业知识。 用于 ECM 可视化和分析的边缘技术，包括二次谐波生成 (SHG) 和 此外，我将利用众多的职业发展资源。