Targeting CC-Chemokine Receptor 7 for the prevention of graft-versus-host disease

靶向 CC-趋化因子受体 7 预防移植物抗宿主病

• 批准号: 8449704
• 负责人: JAMES M COGHILL
• 金额: $ 13.23万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8449704
• 关键词: Accounting; Address; Adverse effects; Allogenic; Animal Model; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Antigens; Attenuated; Biological Assay; Biology; Blocking Antibodies; Blood; Bone Marrow; CC chemokine receptor 7; CCL19 gene; CCL21 gene; Cell Adhesion Molecules; Cell Line; Cell Surface Receptors; Cell physiology; Cells; Cellular biology; Chemicals; Chemotaxis; Clinical; Collaborations; Collection; Color; Complication; Data; Dendritic Cells; Development; Diarrhea; Disease; Disease model; Donor person; Dose; Drug resistance; Drug usage; Effectiveness; Effector Cell; Environment; Exanthema; Exhibits; Funding; Future; Generations; Goals; Hematologic Neoplasms; Hematological Disease; Hematology; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; High Dose Chemotherapy; Human; Immune; Immune response; Immunology; Immunosuppressive Agents; In Vitro; Inflammatory; Infusion procedures; Instruction; K-Series Research Career Programs; Knock-out; Laboratories; Lead; Learning; Ligands; Lymphatic; Lymphocyte; Lymphocyte Homing Receptors; Lymphoid; Lymphoid Tissue; Major Histocompatibility Complex; Malignant - descriptor; Malignant Neoplasms; Mediating; Mentors; Modality; Modeling; Movement; Mus; Nature; Non-Malignant; Normal Cell; Organ; Outcome; Pathogenesis; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Phase; Play; Population; Prevention; Principal Investigator; Process; Property; Protein Kinase; Publishing; Radiation; Reaction; Regimen; Regulatory T-Lymphocyte; Reporter; Research Personnel; Residual Cancers; Role; Running; Secondary to; Siblings; Signal Transduction; Site; Specificity; Spleen; Staging; Stem cell transplant; Stem cells; Steroids; Structure; System; T-Lymphocyte; Therapeutic; Therapeutic procedure; Tissues; Toxic effect; Transplant Recipients; Transplantation; United States National Institutes of Health; Work; base; chemokine receptor; chemotherapy; conditioning; design; didactic education; disorder prevention; drug candidate; drug development; drug discovery; experience; genetically modified cells; graft vs host disease; high throughput screening; human stem cells; immunoregulation; in vivo; irradiation; knockout animal; leukemia; liver injury; lymph nodes; novel; novel strategies; oncology; prevent; receptor; response; responsible research conduct; skills; small molecule; trafficking; tumor

DESCRIPTION (provided by applicant): Allogeneic hematopoietic stem cell transplantation (HSCT) is a therapeutic procedure that is used to treat advanced hematologic malignancies or other blood disorders. HSCT involves the administration of chemotherapy and/or radiation to patients, followed by the transplant of hematopoietic stem cells from a sibling or unrelated matched donor. Because transplant recipients are "rescued" from the bone marrow suppressing effects of the pre-transplant conditioning regimen by the subsequent infusion of healthy stem cells, very high doses of chemotherapy can be administered in an effort to overcome any drug resistance displayed by the malignancy. In addition, mature donor cells present in the stem cell product can mount potent immune responses against any residual cancer that might still be present. Unfortunately, however, these immune reactions are not specific for diseased cells, and normal host tissues can be damaged as well. This process is referred to as graft-versus-host disease (GVHD), and can result in a severe skin rash, profound diarrhea, or liver injury. GVHD is most often treated with high doses of steroids, a therapy that is not always effective and associated with substantial side effects. As a result, laboratories are actively exploring new approaches for treating this difficult disorder. Recently Dr. Coghill, the principal investigatorof the present proposal, published animal model data showing that donor cells lacking the lymph node trafficking receptor CC-Chemokine receptor 7 (CCR7) generate greatly attenuated GVHD responses when transplanted into recipient mice. Cells without CCR7 failed to traffic normally to recipient lymph nodes, and demonstrated a reduced capacity to expand within the spleen. Importantly, however, donor cells lacking CCR7 were able to mount potent anti-cancer immune effects after transplant, and donor regulatory T cells (a population of anti-inflammatory cells) were able to prevent GVHD in the absence of CCR7. Collectively, these findings indicated that blocking the function of CCR7 might prove to be a new and effective approach for preventing GVHD without inhibiting beneficial anti-cancer effects or interfering with the ability of donor regulatory T cells to down-regulate overly aggressive immune responses. There are currently no drugs that block the function of CCR7 that are available for use in HSCT patients or for study in animal models. As a result, this proposal describes a strategy for developing antagonists against CCR7. For this work, we will use genetically engineered cell lines which generate a color signal in the presence of the natural ligands for CCR7: CCL19 and CCL21. With these cells, we will robotically screen a collection of approximately 120,000 small molecules for agents with CCR7 antagonist properties, looking for compounds that are able to block color generation in our primary reporter assay. Actives from the primary screen would then be assessed for cellular toxicity and for specificity against CCR7. Lead compounds would next be evaluated for functionality by assessing their ability to impair chemotaxis (cellular movement) in vitro, a process that is mediated by CCR7. Ultimately, promising compounds would be evaluated for efficacy in vivo by examining their capacity to inhibit GVHD in mouse transplant models. While beyond the scope of the present proposal, lead compounds could be continually refined using medicinal chemistry approaches to maximize their potency and selectivity against CCR7, and developed into drugs for use in human patients undergoing stem cell transplantation. In addition to this CCR7 antagonist discovery effort, mechanistic studies are described to better understand how donor regulatory T cells function after HSCT without CCR7, as previous studies had suggested a critical role for this receptor in their normal function. This proposal is being submitted for a K08 mentored career development award. As a result, the project is designed to provide a substantial educational component for the principal investigator in addition to accomplishing its specific scientific objectives. Dr. Coghill's long-trm goal is to run his own laboratory, with a focus on GVHD pathogenesis and the use of novel, small molecule therapeutics for its treatment and/or prevention. While possessing a strong background in hematology and oncology, this work will offer Dr. Coghill an opportunity to receive additional in-depth instruction in immunology, cell signaling, and the early phases of drug discovery. Dr. Coghill will learn first-hand from an expert mentoring team led by Drs. Jonathan Serody and Stephen Frye, world experts in the fields of lymphocyte trafficking and drug development respectively. A formalized didactic curriculum is also proposed, including coursework in pharmacology, cell biology, grantsmanship, and the responsible conduct of research. Through these experiences, Dr. Coghill will obtain the skills necessary to transition to an independent, NIH funded laboratory investigator.

描述（由申请人提供）：同种异体造血干细胞移植（HSCT）是一种用于治疗晚期血液恶性肿瘤或其他血液疾病的治疗方法。 HSCT 涉及对患者进行化疗和/或放疗，然后移植来自兄弟姐妹或不相关的匹配供体的造血干细胞。由于通过随后输注健康干细胞，移植受者可以从移植前调理方案的骨髓抑制作用中“解救”出来，因此可以施用非常高剂量的化疗，以克服恶性肿瘤所表现出的任何耐药性。此外，干细胞产品中存在的成熟供体细胞可以针对任何可能仍然存在的残留癌症产生有效的免疫反应。然而不幸的是，这些免疫反应并不是针对患病细胞的特异性，正常宿主组织也可能受到损害。这个过程被称为移植物抗宿主病（GVHD），可能导致严重皮疹、严重腹泻或肝损伤。 GVHD 最常采用高剂量类固醇治疗，这种疗法并不总是有效，并且会产生大量副作用。因此，实验室正在积极探索新的 治疗这种困难疾病的方法。 最近，本提案的首席研究员 Coghill 博士发表的动物模型数据显示，缺乏淋巴结运输受体 CC-趋化因子受体 7 (CCR7) 的供体细胞在移植到受体小鼠体内时，会产生大大减弱的 GVHD 反应。没有 CCR7 的细胞无法正常运输至受体淋巴结，并且在脾脏内扩张的能力降低。然而，重要的是，缺乏 CCR7 的供体细胞能够在移植后产生有效的抗癌免疫作用，而供体调节性 T 细胞（一群抗炎细胞）能够在缺乏 CCR7 的情况下预防 GVHD。总的来说，这些发现表明，阻断 CCR7 的功能可能被证明是预防 GVHD 的一种新的有效方法，而不抑制有益的抗癌作用或干扰供体调节性 T 细胞下调过度攻击性免疫反应的能力。 目前尚无阻断 CCR7 功能的药物可用于 HSCT 患者或动物模型研究。因此，该提案描述了开发 CCR7 拮抗剂的策略。在这项工作中，我们将使用基因工程细胞系，在 CCR7 天然配体存在的情况下产生颜色信号：CCL19 和 CCL21。利用这些细胞，我们将自动筛选大约 120,000 个小分子的集合，寻找具有 CCR7 拮抗剂特性的药物，寻找能够在我们的主要报告基因测定中阻止颜色生成的化合物。然后将评估来自初级筛选的活性物质的细胞毒性和针对 CCR7 的特异性。接下来将通过评估先导化合物在体外削弱趋化性（细胞运动）的能力（由 CCR7 介导的过程）来评估其功能。最终，将通过检查它们在小鼠移植模型中抑制 GVHD 的能力来评估有前景的化合物的体内功效。虽然超出了本提案的范围，但可以使用药物化学方法不断精炼先导化合物，以最大限度地提高其针对 CCR7 的效力和选择性，并开发成用于接受干细胞移植的人类患者的药物。除了 CCR7 拮抗剂发现工作之外，还描述了机制研究，以更好地了解在没有 CCR7 的 HSCT 后供体调节性 T 细胞如何发挥作用，因为之前的研究表明该受体在其正常功能中发挥着关键作用。 该提案正在提交以获得 K08 指导的职业发展奖。因此，该项目除了实现其特定的科学目标之外，还旨在为主要研究者提供实质性的教育内容。 Coghill 博士的长期目标是经营自己的实验室，重点研究 GVHD 发病机制以及使用新型小分子疗法进行治疗和/或预防。尽管拥有深厚的血液学和肿瘤学背景，这项工作将为 Coghill 博士提供在免疫学、细胞信号传导和药物发现的早期阶段接受额外深入指导的机会。 Coghill 博士将向由 Drs. Coghill 博士领导的专家指导团队学习第一手资料。乔纳森·塞罗迪（Jonathan Serody）和斯蒂芬·弗莱（Stephen Frye）分别是淋巴细胞贩运和药物开发领域的世界专家。还提出了正式的教学课程，包括药理学、细胞生物学、资助和负责任的研究行为等课程。通过这些经验，Coghill 博士将获得过渡到 NIH 资助的独立实验室调查员。