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体干细胞移植来改善同种异体干细胞移植的结果 丰富供体树突状细胞（DC）群体，减少移植物抗宿主病并提高生存率。 目前的提案基于已发表的研究结果，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重大影响。来自小鼠的临床前数据 BMT 模型系统证明供体浆细胞样树突状细胞 (pDC) 来自骨髓，而非 G-CSF 动员的移植物可防止胸腺所在地的 GvHD，并限制 CCR9-、IL12- 和 IDO- 中的 GvHD 依赖方式。优化移植物中的免疫细胞的问题非常重要，因为患者 移植更多骨髓供体的 pDC 的存活率提高 20%，慢性 GvHD 较少，并且没有 与较少供体 pDC 的受者相比，复发风险增加。未回答的限制性问题 将这些令人兴奋的观察结果应用于广泛的临床实践，包括解决 DC 的异质性 在骨髓和细胞因子动员的移植物中，以确定赋予有利移植的关键子集 结果；定义供体 DC 调节 GvHD 的机制；并确定具有成本效益的程序 用于动员和收获最佳的造血祖细胞移植物，需要最少的离体操作。 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. 明确供体 DC 限制 GvHD 的机制和分子特征。假设：鼠类 人类骨髓中含有出现频率较高的免疫抑制 DC 亚群 与 G-CSF 动员的移植物相比，它抑制 T 细胞响应同种异体抗原的活化，并限制 GvHD。 目标2.增强DC在同种异体移植中的免疫调节能力。假设：治疗 短期 Flt3-L 供体的数量将增加未成熟细胞的数量和免疫效力 供体 DC 限制移植至同种异体受体后的 GvHD。 目标 3. 确定供体来源 DC 的归巢和持久性如何影响 GvHD 和 GvL。 假设：供体 pDC 位于移植受者体内的血液淋巴组织中，通过以下方式支持耐受性： 胸腺中同种异体反应性 T 细胞的阴性选择和/或 Treg 的产生。 拟议的研究将使用单细胞 RNAseq 和质谱流式技术来定义小鼠 DC 簇 以及具有良好免疫特性的人骨髓和细胞因子动员移植物，制定时间表 Flt3-L 给药可增加免疫调节 DC 的频率和效力，并使用生物- 发光和荧光成像追踪小鼠供体 DC 的体内归巢、扩增和功能 骨髓移植模型。这项研究将从机制上理解供体 DC 子集如何调节免疫 同种异体干细胞移植后，将为实体器官移植和过继性 T 细胞研究提供信息 疗法，如 CART。积极的结果可能会导致对干细胞捐赠者进行 Flt3-L 治疗的临床试验。