Engineering Bone Marrow Cells for Immunotherapy

用于免疫治疗的工程骨髓细胞

• 批准号: 7071154
• 负责人: ESMA S YOLCU
• 金额: $ 21.53万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7071154
• 关键词: T lymphocyte; apoptosis; bioengineering /biomedical engineering; biological models; biotechnology; bone marrow; bone marrow transplantation; busulfan; genetic manipulation; histocompatibility; homologous transplantation; immune tolerance /unresponsiveness; immunomodulators; immunosuppressive; immunotherapy; isoantigen; laboratory mouse; leukocyte activation /transformation; sirolimus; surface antigens; therapy design /development; tissue mosaicism; transplantation immunology

DESCRIPTION (provided by applicant): The main objective of this proposal is to use a novel and practical approach to engineer bone marrow cells to display on their surface a modified form of FasL protein with potent apoptotic activity as an immunomodulatory agent and test the capacity of the engineered cells to engraft in allogeneic recipients without complications of graft-versus-host (GVH) or host-versus-graft (HVG) reactions. Allogeneic bone marrow transplantation (BMT) has the potential to cure a series of inherited and acquired hematological disorders and malignancies. BMT can also be used as a cell-based immunomodulatory approach to induce tolerance to foreign and auto-antigens for the prevention and/or treatment of foreign graft rejection and autoimmune disorders. The routine application of BMT as a therapeutic intervention in the clinic, however, is complicated by two different types of immunological reactions; HVG and GVH. HVG reaction is responsible for rejection of allogeneic BM transplants whereas GVH reaction is the major cause of graft versus- host disease (GVHD), which is a life-threatening complication of BMT without effective treatment modalities. T cells specific for alloantigens are the primary culprit in GVH and HVG reactions. Elimination of T cells from the donor BM inoculum or the graft recipient can curtail both types of reactions, lack of engraftment and immunoincompetence are the observed complications. Thus, strategies targeting specific and effective elimination of only the pathogenic T cells may have important implications for routine application of BMT to the clinic for the treatment of a variety of diseases. A novel technique, designated as ProtEx(tm), has recently been developed in our laboratory that allows for rapid (~ 2 hr) and efficient (100% of the targetted cells) display of exogenous proteins of interest on any cell without compromising the function of the cell or the proteins. In this application, ProtEx(tm) will be used to display a modified form of FasL protein with potent apoptotic activity on BM cells and the engineered cells will then be used for transplantation into allogeneic lethally irradiated mice to prevent GVHD and into minimally conditioned hosts to establish mixed chimerism and donor specific tolerance. The use of FasL as an immunomodulatory agent is based on the critical role this molecule plays in immune homeostasis and establishment of self tolerance. We hypothesize that mature T cells in the BM inoculum displaying FasL will respond to the host antigens, upregulate the death receptor Fas, and undergo apoptosis following the engagement of FasL with Fas on the same or a different cell, resulting in the prevention of GVHD. Similarly, recipient T cells reacting to the donor BM cells displaying FasL are expected to undergo apoptosis, leading to engraftment and establishment of mixed chimerism. Rapamycin, an apoptosis-inducing agent, and bisulfan, an alkylating agent that preferentially depletes early hematopoietic stem cells, will be used as adjuvant therapies to i) accentuate FasL-mediated apoptosis, ii) create "space" for donor stem cell engraftment, and iii) establish a clinically applicable protocol. Specific elimination of pathogenic T cells in lethally irradiated or minimally conditioned hosts is expected to results in allogeneic BM engraftment without complications of GVHD and immunoincompetence with strong therapeutic implications.

描述（由申请人提供）：该提案的主要目的是使用一种新颖的方法来工程骨髓细胞在其表面上显示具有有效的凋亡活性的修改形式的FASL蛋白，作为免疫调节剂，并测试工程细胞在同种异体中的工程细胞的能力，而无需Graft-versivers（GRAFT-Versivers）或宿主（HOST）（HOST）（HOST）（HOST）。 同种异体骨髓移植（BMT）具有治愈一系列遗传和获得的血液学疾病和恶性肿瘤的潜力。 BMT也可以用作一种基于细胞的免疫调节方法，以诱导外国和自身抗原的耐受性，以预防和/或治疗外国移植物排斥和自身免疫性疾病。但是，两种不同类型的免疫反应使BMT的常规应用在诊所中作为治疗干预措施变得复杂。 HVG和GVH。 HVG反应负责排斥同种异体BM移植物，而GVH反应是移植与宿主疾病（GVHD）的主要原因，这是BMT的生命危险并发症，没有有效的治疗方式。特异性同种抗原的T细胞是GVH和HVG反应中的主要罪魁祸首。观察到的并发症是消除供体BM接种物中的T细胞或接枝受体的接种者可以减少两种类型的反应，缺乏植入和免疫能力的并发症。因此，针对特定和有效消除病原T细胞的策略可能对BMT在诊所的常规应用以治疗多种疾病具有重要意义。最近在我们的实验室中开发了一种新型技术，该技术被指定为蛋白质（TM），该技术允许在任何细胞上迅速（约2小时）且有效地（100％靶向细胞）显示出感兴趣的外源性蛋白质，而不会损害细胞或蛋白质的功能。在此应用中，Protex（TM）将用于在BM细胞上显示具有有效的凋亡活性的FASL蛋白的改良形式，然后将使用工程细胞用于移植到同种异体辐照的小鼠中，以防止GVHD，以防止GGVHD，并将其转化为最小条件的宿主，以建立混合的Chimerism和供体特定的耐受性。将FASL用作免疫调节剂是基于该分子在免疫稳态和建立自耐力的关键作用。我们假设显示FASL的BM接种物中的成熟T细胞将对宿主抗原作出反应，上调死亡受体FAS，并在FASL与FAS与FAS与FAS互动相同或其他细胞后经历凋亡，从而导致GVHD的预防。同样，有望与表现出FASL的供体BM细胞反应的受体T细胞有望经历凋亡，从而导致植入和建立混合嵌合体。雷帕霉素是一种凋亡诱导剂，而Bisulfan是一种优先耗尽早期造血干细胞的烷基化剂，将用作i）强调Fasl介导的凋亡的辅助疗法，ii）为供体干细胞的“空间”建立了供体干细胞的“空间”。预计致命辐照或最低条件的宿主中病原T细胞的特异性消除会导致同种异体BM植入，而没有GVHD并发症并具有强烈​​的治疗意义。