Proteotranscriptomic regulation of homing and engraftment in hematopoietic stem and progenitor cells for hematopoietic cell transplantation

项目摘要/摘要 迫切需要提高造血干细胞(HSCs)和祖细胞的功能能力 人脐血来源细胞(HPC)增强造血细胞的作用 移植(HCT)用于治疗造血功能紊乱。然而，很少有研究将重点放在 确定参与调节HSC和HPC归巢和植入的全球蛋白质和基因程序 以确定提高红细胞压积的治疗靶点。这项研究中提出的 应用程序将检查二肽的全球蛋白质靶点和依赖于二肽的全球基因程序 多肽酶4(DPP4)是一种调节性丝氨酸蛋白酶，在造血和红细胞压积中发挥重要作用。 我们实验室以前的工作已经证明，DPP4抑制通过增强CB具有临床实用价值 移植治疗造血功能紊乱的患者，尽管这种DPP4抑制必须在高剂量应用 观察不同剂量对脐血红细胞压积的影响。因此，充分了解DPP4的靶点和基因 抑制DPP4导致Hct改善的程序将允许阐明分子 靶向治疗的机制，或者通过靶向DPP4下游的通路来提高疗效 或通过确定可与DPP4结合用于进一步增强Hct的靶向途径。 此外，拟议的研究将检查在HSC和HPC特定亚群中丰富的基因程序 利用多模式单细胞RNA测序技术有效地定位到骨髓 将使我们能够通过细胞表面蛋白的表达和转录组来定义细胞。分析 富含HSC和HPC的基因程序可作为免疫缺陷患者骨髓的归宿 小鼠将允许阐明基因程序，这些程序可以通过抑制或稳定来增强 归位，允许通过新的治疗方式来提高红细胞压积。这项研究计划的目标是 阐明可靶向增强脐带血造血干细胞和造血干细胞归巢和植入的新机制。 研究人员的长期目标是提供对改善HCT的新方式的洞察力 治疗方面，全面提高对造血功能紊乱的治疗水平。成功完成 在此奖学金申请中概述的研究计划和职业发展计划将允许申请者 培养1)分析复杂蛋白质组、转录组和多模式组学数据集的专业知识，2)使用 这些数据用于开发新的方法来改进造血干细胞和造血干细胞的归巢和植入，并 了解造血调节，以及3)测试基因操作或药物治疗对 HCT。这将为申请者管理一个专注于分子的学术实验室做好准备 使用NEXT可用于改善造血障碍患者预后的机制 世代测序、蛋白质组学、基础细胞生物学和生物化学以及小鼠建模方法。

Project Summary/ Abstract There is a strong need to improve the functional capacity of hematopoietic stem cells (HSCs) and progenitor cells (HPCs) derived from human umbilical cord blood (CB) to enhance the efficacy of hematopoietic cell transplantation (HCT) for the treatment of disordered hematopoiesis. However, few studies have focused on defining the global protein and gene programs involved in regulating HSC and HPC homing to and engraftment in the bone marrow in order to identify therapeutic targets to improve HCT. The research proposed in this application will examine the global protein targets of and global gene programs dependent on Dipeptidyl peptidase 4 (DPP4), a regulatory serine protease that plays important roles in hematopoiesis and HCT. Previous work by our lab has demonstrated that DPP4 inhibition has clinical utility by enhancing CB transplantation for patients with disordered hematopoiesis, though this DPP4 inhibition must be applied at high doses to observe modest effects on CB HCT. Thus, fully understanding the targets of DPP4 and the gene programs affected by inhibiting DPP4, which leads to improved HCT, will allow for elucidation of molecular mechanisms to target therapeutically, either by targeting pathways downstream of DPP4 for improved efficacy or by identifying pathways that can be targeted in combination with DPP4 for further enhancement of HCT. Further, the proposed study will examine gene programs enriched in specific subpopulations of HSC and HPC that can efficiently home to the bone marrow utilizing multi-modal single cell RNA-sequencing technology that will allow us to define cells by both their cell surface protein expression and their transcriptome. Analysis of gene programs that are enriched in HSC and HPC that can home to the bone marrow of immune deficient mice will allow for elucidation of gene programs that can be targeted by inhibition or stabilization to enhance homing, allowing for novel therapeutic manners by which to enhance HCT. The goal of this research plan is to elucidate novel mechanisms that can be targeted to enhance homing and engraftment of CB HSCs and HPCs. The investigator's long-term goal is to provide insight into novel manners by which to improve HCT therapeutically for overall improvement of treatment for disordered hematopoiesis. Successful completion of the research plan and career development plan outlined in this fellowship application will allow the applicant to develop expertise in 1) analyzing complex proteomic, transcriptomic, and multi-modal omic data sets, 2) using these data to develop novel approaches to improve homing and engraftment of HSCs and HPCs and to understand hematopoietic regulation, and 3) testing the effects of genetic manipulations or drug treatments on HCT. This will prepare the applicant for a productive career managing an academic lab focused on molecular mechanisms that can be exploited to improve patient outcomes for disordered hematopoiesis using next generation sequencing, proteomics, basic cell biology and biochemistry, and mouse modelling approaches.