Stable shRNA-mediated gene silencing of the beta 7 integrin to ameliorate graft-v

稳定shRNA介导的β7整合素基因沉默可改善移植物v

基本信息

批准号：
8098181
负责人：
Jenny Zilberberg
金额：
$ 22.01万
依托单位：
HACKENSACK UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8098181
关键词：
Acute Acute Graft Versus Host Disease Adhesions Affect Alloantigen Allogenic Biological Assay Biological Preservation Blood Blood donor Bone Marrow Transplantation CD8B1 gene Cell Adhesion Cell Adhesion Molecules Cell Therapy Cell surface Cells Clinical Cytomegalovirus Engineering Event Frequencies Gastrointestinal tract structure Gene Expression Gene Silencing Genetic Techniques Graft-Versus-Tumor Induction Hematologic Neoplasms Hematopoietic stem cells High Endothelial Venule Homing Human Immune Individual Infection Infiltration Integrins Interferon Type II Intervention Intestinal Graft Versus Host Disease Intestines Knockout Mice Ligands Liver Lymphocyte Lymphoid Tissue Malignant - descriptor Malignant Neoplasms Mediating Mesentery Methodology Minor Histocompatibility Antigens Modeling Monoclonal Antibodies Morbidity - disease rate Mus Myeloid Leukemia Opportunistic Infections Organ Patients Peripheral Blood Lymphocyte Play Production Recurrent disease Regimen Regulation Residual Tumors Role SELL gene Selectins Severity of illness Site Skin Specificity Spleen Structure of aggregated lymphoid follicle of small intestine Surface T cell response T memory cell T-Lymphocyte Testing Therapeutic Tissues Translations Transplant Recipients Transplantation chemokine receptor conditioning cytotoxic enzyme linked immunospot assay gastrointestinal epithelium graft vs host disease integrin beta7 knock-down lymph nodes migration mortality mouse model neoplastic cell novel prevent public health relevance receptor reconstitution response small hairpin RNA trafficking tumor

项目摘要

DESCRIPTION (provided by applicant): Adoptive T cell therapy in the form of allogeneic blood and marrow transplantation (BMT) has proven to be one of the few curative treatments for hematological malignancies. Mature donor T cells in the donor inoculum play a central role in mediating graft-versus-tumor (GVT) responses against residual tumor cells that persist after conditioning regimens, and also in facilitating donor immune reconstitution. However the full exploitation of this clinical intervention is greatly limited by the occurrence of graft-versus-host disease (GVHD), which remains one of the main complications of BMT. Acute GVHD is characterized by severe, and potentially lethal, tissue damage to the skin, liver, gastrointestinal tract and lymphoid tissues of transplanted patients, mediated by donor T cells responding to host alloantigens. The homing of T cells to GVHD target organs and their regulation via integrins, selectins and chemokine receptors has therefore been recognized as potential novel sites for intervention to ameliorate or prevent GVHD while still allowing GVT effects. T cell trafficking and homing involves a compendium of events, that require the expression of specific adhesion molecules and chemokine receptors on the T cell surface, along with the spatial and temporal manifestation of their ligand counterparts on high endothelial venules (HEV) of inflamed tissues. Indeed, a number of studies focusing on the manipulation of the ¿7 chain expression of the a4¿7 integrin (an intestinal homing receptor expressed on the surface of T cells) found decreased infiltration of donor T cells to the host gut epithelium, which in turn was associated with decreased intestinal tissue damage in various GVHD murine models. The clinical translation of these studies is however hindered by the lack of an available approach that could safely and efficaciously manipulate T cells in the donor hematopoietic stem cell inoculum, to downregulate their expression of a4¿7 integrin. The proposed studies will thus focus on the use of small-hairpin RNA (shRNA) to test the hypothesis that stable knockdown expression of the ¿7 integrin chain of donor T cells via lentiviral infection, prior to transplant, can provide long-term reduction of GVHD severity without affecting the beneficial GVT potential of those donor T cells. The immune functionality and cell adhesion capabilities of ¿7-shRNA-transduced human T cells will also be assessed, as a first step towards translation of the proposed methodology to the clinical setting. PUBLIC HEALTH RELEVANCE: Graft-versus-host disease (GVHD) still remains one of the main complications associated with bone marrow transplantation (BMT), a well accepted treatment for a number of blood malignancies. In the current study we will investigate the implementation of novel genetic techniques to redirect donor T cells (the causal entities of GVHD) away from inflicting damage to the patient's organs (the hallmark of GVHD). This approach will open up a number of possibilities for new and potentially less aggressive therapeutic strategies for BMT. More importantly, the results from the proposed study could increase the number of donors for BMT, because transplanted T cells would be engineered to infiltrate less into the organs where they can cause undesirable harm upon encountering differences between themselves and the host cells; a situation that occurs specially when the donor cannot be fully matched.

描述（由申请人提供）：同种异体血液和骨髓移植（BMT）形式的过继性 T 细胞疗法已被证明是少数几种治疗血液恶性肿瘤的方法之一，供体接种物中的成熟供体 T 细胞在其中发挥着核心作用。介导针对预处理方案后持续存在的残留肿瘤细胞的移植物抗肿瘤（GVT）反应，并促进供体免疫重建。然而，这种临床干预措施的充分利用是非常重要的。移植物抗宿主病 (GVHD) 的发生仍然是 BMT 的主要并发症之一，急性 GVHD 的特点是对皮肤、肝脏、胃肠道和淋巴组织造成严重且可能致命的组织损伤。因此，T 细胞向 GVHD 靶器官的归巢及其通过整合素、选择素和趋化因子受体的调节被认为是潜在的新疗法。改善 GVHD 的干预位点，同时仍允许 T 细胞运输和归巢，涉及一系列事件，这些事件阻止 T 细胞表面上特定粘附分子和趋化因子受体的表达，以及它们的空间和时间表现。事实上，许多研究都集中在 ¿ 的操作上。 a4¿7整合素（一种在T细胞表面表达的肠道归巢受体）的7链表达发现，供体T细胞对宿主肠上皮的浸润减少，这反过来又与各种GVHD小鼠模型中肠道组织损伤的减少有关然而，由于缺乏一种可以安全有效地操纵供体造血干细胞接种物中的 T 细胞以下调其表达的可用方法，这些研究的临床转化受到阻碍因此，拟议的研究将集中于使用小发夹 RNA (shRNA) 来测试 ¿7 整合素稳定表达的假设。移植前，通过慢病毒感染供体 T 细胞的 7 整合素链，可以长期降低 GVHD 的严重程度，而不影响这些供体 T 细胞的有益 GVT 潜力 ¿还将评估 7-shRNA 转导的人类 T 细胞，作为将所提出的方法转化为临床环境的第一步。公共健康相关性：移植物抗宿主病（GVHD）仍然是与骨髓移植（BMT）相关的主要并发症之一，骨髓移植是许多血液恶性肿瘤的一种公认的治疗方法。在当前的研究中，我们将调查其实施情况。新颖的基因技术可以重定向供体 T 细胞（GVHD 的致病实体），使其免受对患者器官造成的损害（GVHD 的标志），这种方法将为治疗带来多种可能性。更重要的是，拟议研究的结果可能会增加 BMT 捐献者的数量，因为移植的 T 细胞将被设计成更少地渗透到器官中，而这些器官在遇到差异时可能会造成不良伤害。自身和宿主细胞；当供体不能完全匹配时特别发生的情况。