Towards Improving Bone Marrow Transplantation via Inhibition of 15-PGDH

通过抑制 15-PGDH 改善骨髓移植

• 批准号: 10044072
• 负责人: Amar Desai
• 金额: $ 24.9万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-05 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10044072
• 关键词: Back; Biological Assay; Blood Platelets; Bone Marrow; Bone Marrow Cells; Bone Marrow Stem Cell; Bone Marrow Transplantation; CSF3 gene; CXCL12 gene; Cell Compartmentation; Cell Culture Techniques; Cells; Chemosensitization; Clinical; Colony-forming units; Complication; Data; Dinoprostone; Disease; Dose; EP4 receptor; Engraftment; Enzymes; Erythrocytes; Exposure to; Failure; Flow Cytometry; Generations; Genes; Hematopoiesis; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hemorrhage; Homing; Human; IL3 Gene; Immunohistochemistry; In Vitro; Individual; Knockout Mice; Maps; Marrow; Mediating; Mediator of activation protein; Methylcellulose; Modeling; Molecular; Mus; Natural regeneration; PPBP gene; PTPRC gene; Pathway interactions; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Population; Production; Prostaglandin E Receptor; Prostaglandins; Publishing; Recovery; Research Personnel; Role; Science; Signal Pathway; Signal Transduction; Speed; Stem cell transplant; Stromal Cells; Testing; Time; WFDC2 gene; Wild Type Mouse; Xenograft procedure; cell stroma; clinical development; compare effectiveness; curative treatments; cytokine; daughter cell; experience; hematopoietic stem cell niche; high risk; improved; in vivo; infection risk; inhibitor/antagonist; knock-down; knockout gene; mouse model; neutrophil; novel; peripheral blood; receptor; reconstitution; response; skills; small molecule; small molecule inhibitor; standard of care; stem cell population; stem cells; synergism; transcriptome; transcriptome sequencing; transplant model

Project Summary: Hematopoietic stem cell (HSC) transplantation (HST) is a curative treatment for many hematopoietic malignancies. However, a potentially fatal complication of HST is the high risk of infection and bleeding recipients are exposed to during the up to 4 weeks while awaiting regeneration of peripheral blood neutrophils and platelets. I and collaborators published in 2015 in Science the discovery of a small molecule, SW033291, that potently inhibits the prostaglandin degrading enzyme 15-PGDH and markedly speeds recovery from HST. As co-first author, I led studies that showed that treating with SW033291 during murine HST enhances transplanted stem cell homing to the bone marrow niche; markedly accelerates recoveries of neutrophils, platelets, and erythrocytes; and increases the effective dose of a bone marrow graft by 5-fold. I showed these effects are mediated by increased bone marrow PGE2, which acts on EP2 and EP4 PGE2 receptors to induce key hematopoietic cytokines, SCF and CXCL12, in bone marrow stromal cells. In this application I now propose elucidate the full cellular and signaling mediators of SW033291's effect on HST and to examine the interaction of SW033291 with G-CSF, the clinical standard of care for stimulating hematopoietic recovery after human HST. I will first comprehensively identify the molecular mediators of SW033291 effect (using RNA-seq) and identify the bone marrow cells in which these mediators are induced and act (e.g. identifying the novel 15-PGDH positive bone marrow cell that is first targeted by SW033291 and then the downstream cells in which SCF and CXCL12 are markedly induced). I will second interrogate the in vivo function of candidate mediators of SW033291 effect, first by testing the ability of this drug to potentiate HST when candidate SW033291 mediators (e.g. 15-PGDH, CXCL12, SCF, plus two new candidate mediators CXCL4 and CXCL7) are genetically deleted from both donor mouse stem cells and recipient mouse bone marrow stroma. I will further map the differential effects of these mediators on hematopoietic stem cells versus bone marrow stromal cells by examining the ability of SW033291 to potentiate HST when candidate mediators are selectively deleted only in donor HSCs or only in recipient bone marrow stromal cells. Using these models I will further dissect each mediator's role at each of three successive stages of SW033291 effect that are: i) potentiating homing of donor HSCs to the recipient marrow; ii) inducing hematopoietic cytokines in the marrow HSC niche; iii) potentiating stem cell generation of daughter cells. Third, I will examine the interaction of SW033291 with G-CSF, the standard of care used to augment neutrophil recovery in human HSC, following up preliminary data that shows an additive to synergistic effect of SW033291 combined with G-CSF, and characterizing the mechanism of this synergy through identification of target changes in the transcriptome and phosphoproteome of selected bone marrow populations from mice treated with these agents singly or in combination. Through these studies I will stepwise acquire the experience and skills that will launch me as new and fully independent investigator.

项目概要： 造血干细胞（HSC）移植（HST）是一种治疗许多造血干细胞缺乏的有效方法。 恶性肿瘤然而，HST的潜在致命并发症是受者感染和出血的高风险 在等待外周血中性粒细胞和血小板再生的过程中暴露于长达4周的时间。 我和合作者于2015年在《科学》杂志上发表了一种小分子SW 033291的发现， 抑制前列腺素降解酶15-PGDH并显著加速HST的恢复。作为共同第一 作者，我领导的研究表明，在小鼠HST期间用SW 033291治疗可增强移植干细胞 细胞归巢到骨髓龛;显著加速中性粒细胞、血小板和 红细胞;并将骨髓移植物的有效剂量增加5倍。我展示了这些效应 通过增加的骨髓PGE 2介导，其作用于EP 2和EP 4 PGE 2受体以诱导关键的 骨髓基质细胞中的造血细胞因子SCF和CXCL 12。在这个应用程序中，我现在建议 阐明SW 033291对HST作用的完整细胞和信号介质，并检查相互作用 SW 033291与G-CSF（刺激人HST后造血恢复的临床标准治疗）的比较。 我将首先全面鉴定SW 033291效应的分子介质（使用RNA-seq），并鉴定 这些介质在其中被诱导并起作用的骨髓细胞（例如鉴定新的15-PGDH阳性 首先被SW 033291靶向的骨髓细胞，然后是SCF和CXCL 12 明显的诱导）。我将第二次询问SW 033291效应的候选介质的体内功能， 首先通过测试当候选SW 033291介体（例如15-PGDH， CXCL 12、SCF，加上两个新的候选介体CXCL 4和CXCL 7）从两个供体中基因缺失 小鼠干细胞和受体小鼠骨髓基质。我将进一步描绘这些差异的影响， 通过检查SW 033291对造血干细胞与骨髓基质细胞的能力， 当候选介质仅在供体HSC中或仅在受体骨中被选择性删除时， 骨髓基质细胞使用这些模型，我将进一步剖析每个调解人的作用，在每三个连续的 SW 033291作用的阶段是：i）增强供体HSC向受体骨髓的归巢; ii）诱导 骨髓HSC小生境中的造血细胞因子; iii）增强子细胞的干细胞生成。第三、 我将检查SW 033291与G-CSF的相互作用，G-CSF是用于增加中性粒细胞 人HSC中的恢复，随访显示SW 033291协同效应的累加效应的初步数据 联合G-CSF，并通过鉴定靶点变化来表征这种协同作用的机制 在用这些药物治疗的小鼠的选定骨髓群体的转录组和磷酸化蛋白质组中， 试剂单独或组合。通过这些学习，我将逐步获得经验和技能， 让我成为新的独立调查员