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

项目总结/摘要 因此，迫切需要提高造血干细胞（HSC）和祖细胞的功能能力， 来源于人脐带血（CB）的造血干细胞（HPCs）以增强造血干细胞的功效 造血干细胞移植（HCT）用于治疗造血功能紊乱。然而，很少有研究关注 定义参与调节HSC和HPC归巢和植入的全局蛋白质和基因程序 在骨髓中进行研究，以确定改善HCT的治疗靶点。这项研究提出， 应用程序将检查依赖于二肽基的全球蛋白质靶点和全球基因程序 肽酶4（DPP 4），一种在造血和HCT中起重要作用的调节性丝氨酸蛋白酶。 我们实验室以前的工作已经证明，DPP 4抑制通过增强CB具有临床实用性， 因此，对于造血障碍的患者来说，这种DPP 4抑制必须在高浓度下应用。 剂量，以观察对CB HCT的适度影响。因此，充分了解DPP 4和基因的靶点， 受抑制DPP 4影响的程序，这导致了HCT的改善，将允许阐明分子机制。 治疗靶向机制，通过靶向DPP 4下游途径以提高疗效 或者通过鉴定可以与DPP 4组合靶向的途径来进一步增强HCT。 此外，拟议的研究将检查在HSC和HPC的特定亚群中富集的基因程序 利用多模式单细胞RNA测序技术， 将允许我们通过细胞表面蛋白表达和转录组来定义细胞。分析 富含HSC和HPC的基因程序，可以使免疫缺陷患者的骨髓归巢， 小鼠将允许阐明可以通过抑制或稳定来靶向增强的基因程序， 归巢，允许通过其增强HCT的新的治疗方式。这项研究计划的目标是 阐明了可以靶向增强CB HSC和HPC的归巢和植入的新机制。 研究者的长期目标是提供对改善HCT的新方式的深入了解 在治疗上用于整体改善对造血障碍的治疗。成功完成 本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人 培养以下方面的专业知识：1）分析复杂的蛋白质组学、转录组学和多模式组学数据集，2）使用 这些数据开发新的方法来改善HSC和HPC的归巢和植入， 了解造血调节，3）测试基因操作或药物治疗对 HCT。这将为申请人管理一个专注于分子生物学的学术实验室做好准备。 可以利用的机制，以改善患者的结果，为造血功能障碍，使用next 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.