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: 8649079
• 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/8649079
• 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(申请人提供)：异基因造血干细胞移植(HSCT)是一种用于治疗晚期恶性血液病或其他血液疾病的治疗程序。造血干细胞移植涉及对患者进行化疗和/或放射治疗，然后移植来自兄弟姐妹或无血缘关系的匹配捐赠者的造血干细胞。因为移植受者可以通过随后注入健康的干细胞而从移植前调理方案的骨髓抑制效应中被拯救出来，所以可以进行非常高剂量的化疗，以努力克服恶性肿瘤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对可能仍然存在的任何残留癌症产生强大的免疫反应。然而，不幸的是，这些免疫反应并不是针对患病细胞的，正常的宿主组织也可能受到损害。这一过程被称为移植物抗宿主病(GVHD)，可导致严重的皮疹、严重腹泻或肝脏损伤。GVHD最常见的治疗方法是大剂量类固醇，这种疗法并不总是有效的，而且与大量副作用相关。因此，实验室正在积极探索新的 治疗这种难治性疾病的方法。最近，本研究的首席研究员CogHill博士发表的动物模型数据显示，缺乏淋巴转移受体CC-趋化因子受体7(CCR7)的供体细胞移植到受体小鼠体内时，GVHD反应会大大减弱。没有CCR7的细胞无法正常运输到受体淋巴结，并显示出在脾内扩张的能力减弱。然而，重要的是，缺乏CCR7的供体细胞能够在移植后建立强大的抗癌免疫效果，并且供体调节性T细胞(一组抗炎细胞)能够在没有CCR7的情况下预防GVHD。总之，这些发现表明，阻断CCR7的功能可能被证明是一种新的有效方法来预防GVHD，而不会抑制有益的抗癌作用，也不会干扰供者调节性T细胞下调过度侵略性免疫反应的能力。目前还没有阻断CCR7功能的药物可用于HSCT患者或用于动物模型研究。因此，本提案描述了开发针对CCR7的拮抗剂的战略。在这项工作中，我们将使用基因工程细胞系，这些细胞系在CCR7：CCL19和CCL21的天然配体存在的情况下产生颜色信号。有了这些细胞，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，寻找能够在我们的初级报告分析中阻止颜色生成的化合物。然后，将评估来自初级筛查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损害趋化性(细胞运动)的能力来评估其功能性，这一过程是由CCR7介导的。最终，有希望的化合物将通过检查它们在小鼠移植模型中抑制移植物抗宿主病的能力来评估其在体内的疗效。虽然超出了本提案的范围，但先导化合物可以继续使用药物化学方法进行提炼，以最大限度地提高其对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拮抗剂的发现工作外，还描述了机制研究，以更好地了解供体调节性T细胞在没有CCR7的HSCT后如何发挥功能，因为先前的研究表明，这种受体在其正常功能中起着关键作用。这份建议书正在申请K08职业发展指导奖。因此，该项目的设计目的是除了完成其具体的科学目标外，还为首席研究员提供实质性的教育内容。CogHill博士的长期TRM目标是经营自己的实验室，专注于GVHD的发病机制，并使用新型小分子疗法治疗和/或预防GVHD。虽然在血液学和肿瘤学方面有很强的背景，但这项工作将为科希尔博士提供一个机会，在免疫学、细胞信号和药物发现的早期阶段接受额外的深入指导。科希尔博士将从一个由乔纳森·塞罗迪博士和斯蒂芬·弗莱博士领导的专家指导小组那里进行第一手学习。乔纳森·塞罗迪博士和斯蒂芬·弗莱博士分别是淋巴细胞贩运和药物开发领域的世界专家。正式的教学课程也被提出，包括药理学、细胞生物学、授权术和负责任的研究行为的课程。通过这些经历，科格希尔博士将获得过渡到 一名独立的、由NIH资助的实验室研究员。