Using donor dendritic cells to optimize GvHD and GvL in allogeneic stem cell transplantation

使用供体树突状细胞优化同种异体干细胞移植中的 GvHD 和 GvL

• 批准号: 9893083
• 负责人: Edmund K Waller
• 金额: $ 37.87万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893083
• 关键词: Address; Adoptive Cell Transfers; Affect; Alloantigen; Allogenic; Allografting; Antigen-Presenting Cells; Biological Assay; Biological Models; Blood; Blood Component Removal; Bone Marrow; Bone Marrow Transplantation; CCR9 gene; CSF3 gene; Cell Transplantation; Cell Transplants; Cell physiology; Cells; Clinical; Clinical Trials; Communicable Diseases; Cytometry; Data; Dendritic Cells; Donor Selection; Engineering; Frequencies; Future; Generations; Goals; Graft Rejection; Growth; Harvest; Hematopoietic Stem Cell Mobilization; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Heterogeneity; Home environment; Homing; Human; Immune; Immunity; Immunologic Deficiency Syndromes; Immunologic Tests; Immunologics; Immunosuppressive Agents; In Vitro; Incidence; Inflammatory; Interleukin-12; Intervention; Lead; Luciferases; Lymphoid Tissue; Marrow; Medicine; Modeling; Molecular; Molecular Profiling; Mus; Opportunistic Infections; Organ; Organ Transplantation; Outcome; Pathway interactions; Peripheral; Phenotype; Population; Procedures; Property; Publishing; Recurrent disease; Regulatory T-Lymphocyte; Research; Sampling; Schedule; Severities; Site; Solid; Source; Stem cell transplant; T cell therapy; T-Cell Activation; T-Lymphocyte; Techniques; Testing; Thymus Gland; Transgenic Organisms; Transplant Recipients; Transplantation; Work; adaptive immunity; allotransplant; chronic graft versus host disease; clinical practice; cost effective; cytokine; experimental study; fluorescence imaging; graft vs host disease; graft vs leukemia effect; immunoregulation; improved; improved outcome; in vivo; mortality; mortality risk; mouse model; novel; novel strategies; post-transplant; pre-clinical; preclinical study; relapse risk; response; stem cell therapy; transcriptome sequencing

ABSTRACT: The overarching goal is to improve outcomes in allogeneic stem cell transplantation by identifying and enriching donor dendritic cell (DC) populations that reduce graft-versus-host disease and improve survival. The current proposal is grounded on published findings that the content of donor DC in the allograft and in the blood of allo-transplant recipients have a significant impact on clinical outcomes. Preclinical data from murine BMT model systems demonstrate that donor plasmacytoid dendritic cells (pDC) from marrow but not G-CSF mobilized grafts protect against GvHD, home to the thymus, and limit GvHD in an CCR9-, IL12-, and IDO- dependent manner. The question of optimizing immune cells in the graft is highly significant, as patients transplanted with more marrow donor pDC have 20% better survival, with less chronic GvHD, and without increased risk of relapse compared with recipients of fewer donor pDC. Unanswered questions that limit application of these exciting observations to widespread clinical practice include resolving heterogeneity of DC in marrow and cytokine-mobilized grafts to identify the critical subset(s) that confer favorable transplant outcomes; defining the mechanism by which donor DC regulate GvHD; and identifying cost-effective procedures for mobilization and harvesting optimal hematopoietic progenitor grafts requiring minimal ex vivo manipulation. To address these questions, and make progress towards our overall goal, we propose three Specific Aims: Aim 1. To define the mechanisms and molecular profile of donor DCs that limit GvHD. Hypothesis: Murine and human bone marrow contain immunosuppressive DC subset(s) that are present at higher frequencies than in G-CSF-mobilized grafts, that inhibit T cell activation in response to allo-antigen, and that limit GvHD. Aim 2. To enhance the immuno-regulatory potency of DCs in allotransplantation. Hypothesis: Treatment of donors with a short-course of Flt3-L will increase the numbers and immunological potency of immature donor DC that limit GvHD following transplantation to allogeneic recipients. Aim 3. To determine how homing and persistence of donor-derived DC affects GvHD and GvL. Hypothesis: donor pDC that home to hemato-lymphoid tissue in transplant recipients support tolerance via negative selection of allo-reactive T cells in the thymus and/or the generation of Treg. The proposed research will use single cell RNAseq and mass cytometry to define clusters of DC from mouse and human marrow and cytokine-mobilized grafts with favorable immunological properties, develop schedules of Flt3-L administration that increase the frequency and potency of immune-regulatory DC, and use bio- luminescent and fluorescent imaging to track in vivo homing, expansion, and function of donor DC in mouse BMT models. This research will yield a mechanistic understanding of how donor DC subset(s) regulate immunity after allogeneic stem cell transplant that will inform studies in solid organ transplantation and adoptive T cell therapies such as CART. Positive results can lead to a clinical trial of Flt3-L treatment of stem cell donors.

摘要：总体目标是通过识别异基因干细胞移植中的 以及富集减少移植物抗宿主病和提高存活率的供体树突细胞（DC）群体。 目前的建议是基于已发表的发现，即同种异体移植物和移植物中供体DC的含量 同种异体移植受者的血液对临床结果有显著影响。来自小鼠的临床前数据 骨髓移植模型系统证实，来自骨髓的供体浆细胞样树突状细胞（pDC）而不是G-CSF 动员的移植物保护免受GvHD，胸腺的家园，并在CCR 9-，IL 12-和IDO-中限制GvHD。 依赖的方式。优化移植物中免疫细胞的问题非常重要，因为患者 移植更多的骨髓供体pDC的存活率提高20%，慢性GvHD减少， 与较少供体pDC的接受者相比，复发风险增加。无法回答的问题限制了 这些令人兴奋的观察结果在广泛的临床实践中的应用包括解决DC的异质性 在骨髓和骨髓动员的移植物中，以确定提供有利移植的关键子集 结果;定义捐助DC监管GvHD的机制;并确定具有成本效益的程序 用于动员和收获需要最少离体操作的最佳造血祖细胞移植物。 为了解决这些问题，并朝着我们的总体目标取得进展，我们提出了三个具体目标： 目标1.目的探讨供体DC限制GvHD的机制和分子特征。假设：鼠 人骨髓含有以较高频率存在的免疫抑制DC亚群 比G-CSF动员的移植物，抑制T细胞活化应答同种异体抗原，并限制GvHD。 目标2.目的：增强树突状细胞在同种异体移植中的免疫调节作用。假设：治疗 短期Flt 3-L的供体的数量和免疫效力将增加未成熟的 供体DC限制移植到同种异体受体后的GvHD。 目标3.确定供体来源DC的归巢和持续性如何影响GvHD和GvL。 假设：在移植受体中归巢到血液淋巴组织的供体pDC通过以下途径支持耐受： 胸腺中同种异体反应性T细胞的阴性选择和/或Treg的产生。 这项研究将使用单细胞RNAseq和质谱细胞术来确定小鼠DC的簇 以及具有良好免疫特性的人骨髓和骨髓动员移植物， Flt 3-L给药，增加免疫调节DC的频率和效力，并使用生物- 用于追踪小鼠中供体DC的体内归巢、扩增和功能的发光和荧光成像 BMT模型。这项研究将产生一个机制的理解如何供体DC亚群调节免疫 在同种异体干细胞移植后，这将为实体器官移植和过继性T细胞移植研究提供信息。 治疗方法如CART。阳性结果可以导致Flt 3-L治疗干细胞供体的临床试验。