Targeting Notch2 in Hematopoietic Cell Therapy.

造血细胞治疗中的靶向 Notch2。

• 批准号: 10189683
• 负责人: Lan Zhou
• 金额: $ 59.34万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-12-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10189683
• 关键词: Adhesions; Adhesives; Affinity; Amino Acids; Apoptotic; Attenuated; Binding; Biology; Blood; Bone Marrow; CD34 gene; Cell Adhesion; Cell Communication; Cell Surface Receptors; Cell Therapy; Cell surface; Cells; Clinical; Defect; Disaccharides; Disease; Dose; Endothelial Cells; Endothelium; Engraftment; Epidermal Growth Factor; Extracellular Domain; Failure; Fucose; Funding; Gastroenterology; Genetic; Growth; Health; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Mobilization; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Homing; Human; Inflammation; Infusion procedures; Knowledge; Ligand Binding; Ligands; Location; Maintenance; Malignant - descriptor; Marrow; Modification; Mus; Myeloproliferative disease; Non-Malignant; Occupations; Oncogenic; Osteoblasts; Outcome; Patient-Focused Outcomes; Patients; Peptides; Physiological; Post-Translational Protein Processing; Protein Isoforms; Reagent; Recombinants; Recovery; Regimen; Regulation; Reporting; Research; Resources; Risk; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Stress; Structure; Supporting Cell; Testing; Threonine; Transactivation; Transferase; Transplantation; anticancer research; base; biological adaptation to stress; carcinogenesis; curative treatments; endoplasmic reticulum stress; hematopoietic cell transplantation; hematopoietic engraftment; hematopoietic stem cell niche; hematopoietic stem cell quiescence; improved; improved outcome; leukemia; leukemia initiating cell; mitochondrial metabolism; mouse model; notch protein; novel; pre-clinical; receptor; reconstitution; response; stem cell engraftment; stem cell niche; stem cell self renewal; stem cell survival; stem cell therapy; stem cells; stemness

R01HL103827-06A1 Resubmission (Zhou, Lan) 10.22.2018 Abstract Hematopoietic stem cell (HSC) transplantation is the only curative option for various malignant and a few nonmalignant diseases. A successful outcome is dependent on infusing an adequate number of functionally active mobilized hematopoietic progenitor cells (HPCs). Inadequate mobilization in patients showing poor responses to current mobilizing agents remains a clinical problem. In addition, transplantation using lower-then-desired doses of stem cells increases risks of stem cell engraftment failure. Hence, developing more efficacious and low-risk HPC mobilization regimens and strategies will greatly improve patient outcomes. We recently identified a novel role of Notch as a cell surface adhesion as well as signaling molecule to retain stem cell in the bone marrow microenvironment. Notch is a conserved cell surface receptor that regulates stemness, growth, and differentiation. Notch transactivation is the result of functional engagement of Notch receptors with Notch ligands, which is dependent on the posttranslational modification of Notch receptors with O-fucose and Fringe on the epidermal growth factor (EGF) modules of Notch extracellular domain. Structural studies have demonstrated that O-fucose attached to the specific threonine residue of the core ligand binding region of Notch EGF repeats functions as a surrogate amino acid to make specific and functional contact to Notch ligand DLL4 and JAG1, respectively. Elongation of O-fucose to a disaccharide (GlcNAcβ1- 3Fucose) by Fringe further increases Notch binding affinity to some Notch ligands. In the last funding cycle, we identified conservative Notch-ligand adhesive interaction and its modification by O- fucosylation for HSC quiescence and niche maintenance. We provided compelling evidences showing that deficiency of O-fucosylation leads to decreased HSC quiescence and HSC adhesion to bone marrow niche cells, and increased HSC egress from the marrow. We also reported that neutralizing DLL4 or JAG1 as well as blocking Notch2 significantly enhances HSC egress to the periphery. More, recently, we found transient Notch2 blockade results in superior engraftment and hematopoietic recovery of mobilized HSPC associated with enhanced HSPC activity and attenuated ER stress activation. Further, we found that Fringe-modified recombinant Notch peptides bearing the core ligand binding EGF repeats function as decoys for Notch ligand to induce HSC and progenitor cell egress in osteoblastic spheroids. We thus hypothesize that Notch2 interaction with DLL4 or JAG1 regulates ligand- and niche-specific HSPC retention. We further hypothesize that Notch2-targeted mobilization regimen may improve HCT outcome by mitigating ER stress response activation. We will test this hypothesis in three aims. In Aim 1, we will examine the differential roles of JAG1 and DLL4 expressed by the endothelial and the immature osteolineage cells in the regulation of HSC niche retention, location and mobilization. In Aim 2, we will assess the ligand binding affinity and the efficiency of Fringe-modified Notch peptides as decoys to enhance HSPC egress from HSC niche. In Aim 3, we will examine the significance and the mechanism by which blocking Notch2 potentiates HSPC activity through attenuating ER stress response. At the end of this study, we hope our findings will reveal novel aspects of Notch signaling in HSC niche biology and in ER stress regulation which may provide a resource for understanding Notch signaling in the control of HSC activity during stress hematopoiesis, with implications for identifying ways to improve HCT outcomes.

R01HL103827-06A1 重新提交（周兰）10.22.2018 抽象的 造血干细胞（HSC）移植是治疗多种恶性肿瘤的唯一治疗选择 少数非恶性疾病的成功取决于输注足够数量的药物。 功能活跃的造血祖细胞（HPC）动员不足。 此外，对当前动员剂的反应不佳仍然是一个临床问题。 使用低于所需剂量的干细胞进行移植会增加干细胞植入的风险 因此，开发更有效、低风险的 HPC 动员方案和策略将会失败。 我们最近发现了 Notch 作为细胞表面的新作用。 粘附以及信号分子将干细胞保留在骨髓微环境中。 是一种保守的细胞表面受体，调节干性、生长和分化。 反式激活是 Notch 受体与 Notch 配体功能性结合的结果，即 依赖于Notch受体的O-岩藻糖和Fringe的翻译后修饰 Notch细胞外域的表皮生长因子（EGF）模块的结构研究。 O-岩藻糖附着在核心配体结合区的特定苏氨酸残基上 Notch EGF 重复序列充当替代氨基酸，以进行特异性和功能性接触 Notch 配体 DLL4 和 JAG1，分别将 O-岩藻糖延伸为二糖 (GlcNAcβ1-)。 Fringe 的 3Fucose）进一步增加了 Notch 与某些 Notch 配体的结合亲和力。 循环中，我们确定了保守的Notch-配体粘附相互作用及其通过O-的修饰 我们提供了令人信服的证据表明 HSC 静止和生态位维持的岩藻糖基化。 O-岩藻糖基化的缺陷导致 HSC 静止状态和 HSC 与骨的粘附减少 我们还报道了中和作用。 DLL4 或 JAG1 以及阻塞 Notch2 显着增强了 HSC 到外围的出口。 最近，我们发现短暂的 Notch2 阻断可导致更好的植入和造血 动员的 HSPC 的恢复与 HSPC 活性增强和 ER 应激减弱相关 此外，我们发现Fringe修饰的重组Notch肽带有核心配体。 结合 EGF 重复序列作为 Notch 配体的诱饵，诱导 HSC 和祖细胞排出 因此，我们发现 Notch2 与 DLL4 或 JAG1 相互作用进行调节。 我们进一步研究了Notch2靶向动员。 方案可以通过减轻 ER 应激反应激活来改善 HCT 结果，我们将对此进行测试。 三个目标的假设 在目标 1 中，我们将检查 JAG1 和 DLL4 表达的差异作用。 通过内皮细胞和未成熟骨系细胞调节 HSC 生态位保留， 在目标 2 中，我们将评估配体结合亲和力和效率。 边缘修饰的 Notch 肽作为诱饵来增强 HSPC 从 HSC 生态位的流出。在目标 3 中，我们将。 研究阻断 Notch2 增强 HSPC 活性的意义和机制 通过减弱 ER 应激反应 在本研究结束时，我们希望我们的发现能够揭示新颖的内容。 HSC 生态位生物学和 ER 应激调节中的 Notch 信号传导方面可能提供 了解应激造血过程中 HSC 活性控制中的 Notch 信号传导的资源， 对于确定改善 HCT 结果的方法具有重要意义。