Wnt signaling in hematopoietic development

造血发育中的 Wnt 信号传导

• 批准号: 9220216
• 负责人: David Traver
• 金额: $ 52.34万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9220216
• 关键词: Affinity; Alleles; Anemia; Animal Model; Area; Autoimmunity; Binding; Biochemical; Biochemistry; Biological; Biological Assay; Biology; Blood; Blood Cells; Bone Marrow; Bone Marrow Transplantation; CRISPR/Cas technology; Cell Differentiation process; Cell Lineage; Cell Proliferation; Cell Therapy; Cells; ChIP-seq; Clinic; Cues; Data; Derivation procedure; Development; Developmental Biology; Developmental Process; Disease; Embryo; Endothelial Cells; Endothelium; Engineering; Epigenetic Process; Event; Fishes; Gene Expression; Genes; Genetic; Goals; Hematological Disease; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Human; Immune; Instruction; Knock-out; Life; Medicine; Methods; Molecular; Motivation; Outcome; Pathway interactions; Patients; Phenotype; Play; Pluripotent Stem Cells; Principal Investigator; Process; Property; Protein Biochemistry; Protocols documentation; Regulation; Research; Role; Series; Signal Transduction; Source; Specificity; Technology; Therapeutic; Tissue Engineering; Tissues; Transplantation; WNT Signaling Pathway; WNT9A gene; Wnt proteins; Work; Zebrafish; base; early embryonic stage; effective therapy; experimental study; extracellular; human pluripotent stem cell; improved; in vivo; insight; interest; knock-down; leukemia; next generation sequencing; overexpression; pluripotency; receptor; reconstitution; skills; stem cell biology; transcriptome sequencing

PROJECT SUMMARY Hematopoietic stem cells (HSCs) give rise to all terminally differentiated cells in the blood. The ability of HSCs to reconstitute these blood cell lineages for life underlies the efficacy of bone marrow transplantation therapy for treatment of various blood disorders, including leukemias, anemia, and autoimmunity. Although this is an established and effective treatment, two-thirds of patients in need of a transplant lack a matched donor. Therefore, alternative sources of therapeutic HSCs would be a boon to the field. Human pluripotent stem cells (hPSCs) represent a potential source for cell-based therapies, including the derivation of patient-specific transplantable HSCs, which would additionally circumvent immune rejection and alloreactivity, both major issues in the clinic. The proposed collaborative research leverages the expertise of two Principal Investigators with complementary research interests and skills in stem cell biology, zebrafish genetics and development, hematopoietic development, and Wnt biology and biochemistry. Using zebrafish as a model organism, they seek to identify and characterize the molecular cues, in particular Wnt signaling, that direct hematopoietic development during early embryonic stages. As indicated by their ongoing collaborative studies, the Wnt signaling pathway plays a critical role in the development of HSCs. The proposed studies will build on the observation that Wnt9a is specifically required for HSC formation, emergence and expansion by (1) determining the cognate Wnt9a receptor(s) by genetic, cell biological and biochemical means, (2) generating knockout and conditional knockout alleles of Wnt9a and candidate receptors and thereby assessing their spatial and temporal requirements, (3) identifying the Wnt producing and Wnt-responding cells, and (4) dissecting the gene expression and epigenetic changes that accompany HSC development. The long-term goal of these studies is to gain a better understanding of how HSCs develop in the embryo. Given that fundamental developmental processes, including blood formation, are highly conserved from fish to humans, the discoveries made in the course of the proposed work will directly inform current methods and protocols to derive HSCs from hPSCs. Therefore, successful completion of this research will have a profound impact on HSC derivation and expansion, and thereby will be instrumental in overcoming current obstacles to the effective treatment of diseases requiring bone marrow transplant therapy.

项目总结 造血干细胞(HSCs)是血液中所有终末分化细胞的来源。造血干细胞的能力 重建这些血细胞谱系是骨髓移植治疗效果的基础。 用于治疗各种血液疾病，包括白血病、贫血和自身免疫。虽然这是一个 尽管已经建立了有效的治疗方法，但三分之二需要移植的患者缺乏匹配的捐赠者。 因此，替代的治疗性造血干细胞来源将是该领域的福音。人多能干细胞 (HPSCs)代表了基于细胞的治疗的潜在来源，包括衍生患者特有的 可移植的造血干细胞，这将额外绕过免疫排斥和同种异体反应，这两个主要 诊所里的问题。 拟议的合作研究利用了两名首席调查员的专业知识，具有互补性 干细胞生物学、斑马鱼遗传和发育、造血学的研究兴趣和技能 发展，以及WNT生物学和生物化学。利用斑马鱼作为模式生物，他们试图识别 并描述了指导造血发育的分子信号，特别是Wnt信号 早期胚胎阶段。他们正在进行的合作研究表明，Wnt信号通路在 在肝星状细胞发育中的关键作用。拟议的研究将建立在观察到WNT9A是 对于HSC的形成、出现和扩展是特别需要的，通过(1)确定同源WNT9a 受体(S)通过遗传、细胞生物学和生化手段，(2)产生敲除和条件性 Wnt9a和候选受体的敲除等位基因及其空间和时间的评估 需求，(3)鉴定产生和响应Wnt的细胞，以及(4)解剖基因 伴随着HSC发育的表达和表观遗传学变化。 这些研究的长期目标是更好地了解造血干细胞是如何在胚胎中发育的。 鉴于基本的发育过程，包括血液形成，从鱼类到 人类，在拟议的工作过程中所做的发现将直接告知当前的方法和 从hPSC派生HSC的协议。因此，成功完成这项研究将具有深远的意义 对HSC派生和扩展的影响，从而将有助于克服目前的障碍 需要骨髓移植治疗的疾病的有效治疗。