Wnt signaling in hematopoietic development

造血发育中的 Wnt 信号传导

• 批准号: 9886256
• 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 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9886256
• 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 Therapy; Cells; ChIP-seq; Clinic; Cues; Data; Derivation procedure; Development; Developmental Biology; Developmental Process; Disease; Embryo; Endothelial Cells; Engineering; Epigenetic Process; Event; Fishes; Gene Expression; Genes; Genetic; Goals; Hematological Disease; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic System; Hematopoietic stem cells; Human; Immune; Instruction; Knock-out; Life; Medicine; Methods; Molecular; Motivation; Outcome; Pathway interactions; Patients; Phenotype; Play; Principal Investigator; Process; Property; Protein Biochemistry; Protocols documentation; Regulation; Research; Role; Series; Signal Transduction; Source; Specificity; Stem Cell Development; Therapeutic; Tissue Engineering; Tissues; Transplantation; WNT Signaling Pathway; WNT9A gene; Wnt proteins; Work; Zebrafish; base; blood formation; conditional knockout; early embryonic stage; effective therapy; experimental study; extracellular; hematopoietic stem cell expansion; hematopoietic stem cell formation; hemogenic endothelium; human pluripotent stem cell; human stem cells; improved; in vivo; insight; interest; knock-down; leukemia; next generation sequencing; overexpression; pluripotency; receptor; reconstitution; skills; stem cell biology; stem cell proliferation; 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.

项目摘要 造血干细胞（HSCs）在血液中产生所有终末分化细胞。HSC的能力 重建这些血细胞谱系以维持生命是骨髓移植治疗有效性的基础 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 在现有的有效治疗方法中，三分之二需要移植的患者缺乏匹配的供体。 因此，治疗性HSC的替代来源将是该领域的布恩。人多能干细胞 人PSC（hPSC）代表了基于细胞的疗法的潜在来源，包括患者特异性造血干细胞的衍生。 可移植的HSC，这将另外规避免疫排斥和同种异体反应性，两者都是主要的 诊所中的问题。 拟议的合作研究利用了两位主要研究者的专业知识， 研究兴趣和技能在干细胞生物学，斑马鱼遗传学和发展，造血 发展，Wnt生物学和生物化学。他们以斑马鱼为模式生物， 并表征在造血发育过程中指导造血发育的分子线索，特别是Wnt信号传导， 早期胚胎阶段正如他们正在进行的合作研究所表明的那样，Wnt信号通路在细胞内起着重要的作用。 在造血干细胞的发育中起着关键作用。拟议的研究将建立在Wnt 9a是 HSC形成、出现和扩增所特别需要的，通过（1）确定同源Wnt9a 通过遗传、细胞生物学和生物化学手段，（2）产生敲除和条件性的 敲除Wnt9a和候选受体的等位基因，从而评估它们的空间和时间 （3）鉴定Wnt产生细胞和Wnt应答细胞，以及（4）剖析Wnt产生细胞的基因， 表达和表观遗传变化伴随HSC的发展。 这些研究的长期目标是更好地了解HSC如何在胚胎中发育。 鉴于基本的发育过程，包括血液形成，从鱼类到哺乳动物都是高度保守的， 人类，在拟议的工作过程中取得的发现将直接告知当前的方法， 从hPSC衍生HSC的方法。因此，本研究的成功完成将产生深远的影响 影响HSC的衍生和扩展，从而将有助于克服目前的障碍， 有效治疗需要骨髓移植治疗的疾病。