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（由适用提供）：以同种异体血液和骨髓移植（BMT）的形式使用的养育T细胞疗法已被证明是血液学恶性肿瘤的少数治疗疗法之一。供体接种物中的成熟供体T细胞在调节治疗方案后持续存在的残留肿瘤细胞以及支持供体免疫重构基础上的残留肿瘤细胞中介导移植物肿瘤（GVT）反应中起着核心作用。然而，这种临床干预的全部开发受到移植物抗宿主病（GVHD）的发生的极大限制，该疾病仍然是BMT的主要并发症之一。急性GVHD的特征是受移植患者的皮肤，肝脏，胃肠道和淋巴组织的严重且可能致命的，组织损害，由供体T细胞介导的供体T细胞介导的宿主同种剂。因此，T细胞将T细胞归为GVHD靶器官及其通过整联蛋白，Selectins和趋化因子受体的调节被认为是潜在的新型部位，以改善或预防GVHD，同时仍允许GVT效应。 T细胞运输和归巢涉及事件的汇编，这需要在T细胞表面表达特定的粘附分子和趋化因子受体，以及其在炎症组织的高内皮静脉（HEV）上配体的空间和临时表现。实际上，许多研究重点是操纵``77整合蛋白的链链表达（在T细胞表面表达的肠道寄养受体）的7链表达发现，发现供体T细胞浸润改善了宿主肠肠上皮的浸润，这反过来又与各种GVHD Murine模型中改善的肠道组织损害相关。然而，由于缺乏可以安全有效地操纵供体造血干细胞接种物中T细胞的可用方法，这些研究的临床翻译受到了阻碍，从而下调了它们的A4€7整合蛋白的表达。因此，拟议的研究将集中于使用小发蛋白RNA（SHRNA）来检验以下假设：在移植之前，通过慢病毒感染稳定地敲低的供体T细胞的整合素链链链，可以长期降低GVHD的严重性，而不会影响那些供体T细胞的BENOFISE GVT潜力。还将评估7-shRNA转导的人T细胞的免疫功能和细胞粘附能力，作为将拟议方法转化为临床环境的第一步。公共卫生相关性：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，这是对许多血液恶性肿瘤的公认治疗方法。在当前的研究中，我们将研究新型遗传技术的实施，以重定向供体T细胞（GVHD的因果实体），从对患者器官（GVHD的标志）造成损害。这种方法将为BMT提供新的和潜在的积极性理论策略的许多可能性。更重要的是，拟议研究的结果可能会增加BMT的供体数量，因为移植的T细胞将被设计成较少的渗透到器官中，在这些器官中遇到自身与宿主细胞之间的差异时会造成不良伤害。当捐赠者无法完全匹配时，这种情况是特别发生的情况。