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

• 批准号: 7991155
• 负责人: Jenny Zilberberg
• 金额: $ 26.42万
• 依托单位: HACKENSACK UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7991155
• 关键词: Acute; Acute Graft Versus Host Disease; Adhesions; Affect; Alloantigen; Allogenic; Biological Assay; Biological Preservation; Blood; Blood donor; Bone Marrow Transplantation; 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

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T细胞治疗已被证明是治疗血液系统恶性肿瘤的少数治愈方法之一。在供体接种物中，成熟的供体T细胞在介导移植物抗肿瘤（GVT）反应中起着核心作用，这些反应针对的是调理方案后持续存在的残余肿瘤细胞，同时也促进了供体免疫重建。然而，这种临床干预的充分利用受到移植物抗宿主病（GVHD）的发生的极大限制，GVHD仍然是BMT的主要并发症之一。急性GVHD的特点是移植患者的皮肤、肝脏、胃肠道和淋巴组织受到严重的、潜在致命的组织损伤，由供体T细胞对宿主异体抗原产生反应介导。因此，T细胞归巢到GVHD靶器官，并通过整合素、选择素和趋化因子受体进行调控，已被认为是改善或预防GVHD的潜在新干预位点，同时仍然允许GVT的作用。T细胞运输和归巢涉及一系列事件，这些事件需要在T细胞表面表达特异性粘附分子和趋化因子受体，以及它们在炎症组织的高内皮小静脉（HEV）上的配体对应物的空间和时间表现。事实上，许多关注a4¿7整合素（一种在T细胞表面表达的肠道归巢受体）的7链表达的研究发现，供体T细胞向宿主肠道上皮的浸润减少，这反过来又与各种GVHD小鼠模型中肠道组织损伤减少有关。然而，由于缺乏一种安全有效地操纵供体造血干细胞接种中的T细胞下调其a4¿7整合素表达的方法，这些研究的临床转化受到了阻碍。因此，拟议的研究将侧重于使用小发夹RNA （shRNA）来验证一种假设，即在移植前通过慢病毒感染稳定地敲低供体T细胞的¿7整合素链表达，可以在不影响供体T细胞有益的GVT潜力的情况下长期降低GVHD严重程度。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，作为将所提议的方法转化为临床环境的第一步。