Peripheral T Cell tolerance by targeting AKT

通过靶向 AKT 实现外周 T 细胞耐受

• 批准号: 7842641
• 负责人: CLAUDIO ANASETTI
• 金额: $ 20.88万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-15 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7842641
• 关键词: 1-Phosphatidylinositol 3-Kinase; Acute Graft Versus Host Disease; Alloantigen; Allogeneic Bone Marrow Transplantation; Antigens; Apoptosis; Benign; Bone Marrow Transplantation; Cell Transplantation; Clinic; Clinical; DNA biosynthesis; Data; Development; Disease; Early treatment; Equilibrium; Generations; Goals; Grant; Growth; Hematopoietic; Human; Immune; Immunology; In Vitro; Knock-in Mouse; Knock-out; Laboratories; Literature; Malignant - descriptor; Marrow; Mediating; Methods; Modeling; Morbidity - disease rate; Mus; Nude Mice; Pathway interactions; Patients; Peripheral; Pharmacotherapy; Phosphorylation; Pre-Clinical Model; Prevention; Production; Protein-Serine-Threonine Kinases; Proto-Oncogene Proteins c-akt; Regulation; Regulatory T-Lymphocyte; Rest; Stem cells; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Thymocyte Development; Transgenic Organisms; Transplantation; base; cancer cell; clinically relevant; cytokine; design; disability; disorder prevention; efficacy testing; glucose metabolism; graft vs host disease; graft vs leukemia effect; in vivo; inhibitor/antagonist; mortality; novel; pathogen; pre-clinical; prevent; public health relevance; reconstitution; research study; response; small molecule; trafficking

DESCRIPTION (provided by applicant): The goal of our laboratory is to develop methods for the induction of peripheral T cell tolerance and prevention of graft-versus-host disease (GVHD) after human hematopoietic cell transplantation (HCT). Transplantation can cure a variety of benign and malignant hematopoietic disorders, but GVHD remains the primary case of transplant-related morbidity, disability and mortality, thereby limiting the use of HCT. Donor T cells that are included in the stem cell inoculum and recognize recipient alloantigens mediate GVHD. An ideal approach to prevent GVHD is to selectively deplete the alloactivated T cells with a short course of treatment early after transplantation, while sparing other T cells that would be responsible for immune reconstitution and control of pathogens. Depleting alloreactive T cells may tip the balance towards regulation and tolerance, if regulatory T cells (Tregs) are also spared. We have evaluated the effects of TCN on T cell activation and found that TCN suppresses DNA synthesis, proliferation and promotes apoptosis in vitro. Based on these preliminary data and the evidence in literature that AKT promotes T cell activation, survival, Th1 cytokine production but suppresses Treg generation, we reason that treatment of TCN would prevent the development of GVHD after allogeneic BMT. This grant would generate the required preliminary data for testing TCN for GVHD prevention in patients undergoing allogeneic BMT. If the proposed approach is effective in preventing GVHD in humans, more patients would benefit from BMT and achieve cure. PUBLIC HEALTH RELEVANCE: AKT, a phosphatidylinositol 3-kinase (PI3K)-dependent serine-threonine kinase, promotes glucose metabolism and survival of activated T cells and limits differentiation of regulatory T cells (Tregs). While AKT is critical for thymocyte development, some authors have proposed that AKT is not essential for survival of resting T cells. These functions of AKT make it an attractive target in controlling GVHD. The objective of this proposal is to control acute GVHD in pre- clinical models of allogeneic bone marrow transplantation (BMT) by targeting the AKT pathway. A small molecule AKT inhibitor, triciribine (TCN), suppress the phosphorylation of AKT thereby inhibiting its activity. Treatment with TCN potently inhibits growth in nude mice of human cancer cells that over-express AKT. We plan to develop sufficient preclinical data to allow the design of a clinical GVHD prevention trial in humans. Our central hypothesis is that treatment of TCN suppresses T cell activation, expansion and survival while promoting Treg generation in response to alloantigen in vivo, and thus prevents GVHD development after allogeneic BMT. To test this hypothesis, we will pursue the following two Specific Aims: 1) To determine the effects of TCN on GVHD, immune reconstitution and GVL activity after allogeneic BMT. Our preliminary data indicate that TCN suppresses T cell response to alloantigen and promotes apoptosis of antigen-activated T cells in vitro, suggesting that treatment of TCN may prevent GVHD after allogeneic BMT in vivo. In this Specific Aim, we will test the efficacy of TCN in the prevention of GVHD in pre-clinical murine models of BMT. Furthermore, we will evaluate the effects of TCN on marrow reconstitution and GVL activity, the beneficial effects of donor T cells. 2) To define the mechanisms of TCN in the prevention of GVHD. Multiple mechanisms may be involved in the prevention of GVHD by TCN. In this Specific Aim, we will evaluate the potential effects of TCN on T cell expansion, apoptosis, cytokine production, trafficking and Treg generation, which are critical factors in the development of GVHD. To accomplish these objectives, our team has expertise in immunology, bone marrow transplantation and drug therapy. Unique transgenic, knock-out and knock-in mice are available for all the planned experiments. The proposed studies will definitely evaluate the efficacy and mechanisms of TCN in controlling GVHD in clinically relevant models of BMT, and may point to a novel and more selective strategy to prevent GVHD in the clinic.

描述（由申请人提供）：我们实验室的目标是开发在人类造血细胞移植（HCT）后诱导外周T细胞耐受和预防移植物抗宿主病（GVHD）的方法。移植可以治愈多种良性和恶性造血疾病，但 GVHD 仍然是移植相关发病、残疾和死亡率的主要病例，从而限制了 HCT 的使用。干细胞接种物中包含的供体 T 细胞可识别介导 GVHD 的受体同种抗原。预防 GVHD 的理想方法是在移植后早期通过短期治疗选择性地消耗同种异体激活的 T 细胞，同时保留负责免疫重建和病原体控制的其他 T 细胞。如果调节性 T 细胞 (Treg) 也能幸免，耗尽同种反应性 T 细胞可能会导致调节和耐受的平衡发生变化。我们评估了TCN对T细胞活化的影响，发现TCN在体外抑制DNA合成、增殖并促进细胞凋亡。基于这些初步数据和文献证据表明 AKT 促进 T 细胞活化、存活、Th1 细胞因子产生，但抑制 Treg 生成，我们推断 TCN 治疗将阻止同种异体 BMT 后 GVHD 的发生。这笔赠款将为接受同种异体 BMT 的患者测试 TCN 预防 GVHD 所需的初步数据。如果所提出的方法能有效预防人类 GVHD，更多患者将受益于 BMT 并实现治愈。 公共健康相关性：AKT 是一种磷脂酰肌醇 3 激酶 (PI3K) 依赖性丝氨酸-苏氨酸激酶，可促进葡萄糖代谢和活化 T 细胞的存活，并限制调节性 T 细胞 (Treg) 的分化。虽然 AKT 对于胸腺细胞发育至关重要，但一些作者提出 AKT 对于静息 T 细胞的存活并不是必需的。 AKT 的这些功能使其成为控制 GVHD 的一个有吸引力的靶标。该提案的目的是通过靶向 AKT 通路来控制同种异体骨髓移植 (BMT) 临床前模型中的急性 GVHD。小分子 AKT 抑制剂曲西瑞滨 (TCN) 可抑制 AKT 的磷酸化，从而抑制其活性。 TCN 治疗可有效抑制裸鼠体内过度表达 AKT 的人类癌细胞的生长。我们计划开发足够的临床前数据，以便设计人类临床 GVHD 预防试验。我们的中心假设是，TCN 治疗可抑制 T 细胞活化、扩增和存活，同时促进体内同种异体抗原应答的 Treg 生成，从而防止同种异体 BMT 后发生 GVHD。为了检验这一假设，我们将追求以下两个具体目标： 1) 确定 TCN 对同种异体 BMT 后 GVHD、免疫重建和 GVL 活性的影响。我们的初步数据表明，TCN 抑制 T 细胞对同种异体抗原的反应，并在体外促进抗原激活的 T 细胞凋亡，这表明 TCN 治疗可以预防体内同种异体 BMT 后的 GVHD。在此具体目标中，我们将在临床前 BMT 小鼠模型中测试 TCN 在预防 GVHD 方面的功效。此外，我们将评估 TCN 对骨髓重建和 GVL 活性的影响，以及供体 T 细胞的有益作用。 2) 明确TCN预防GVHD的机制。 TCN 预防 GVHD 可能涉及多种机制。在此具体目标中，我们将评估 TCN 对 T 细胞扩增、凋亡、细胞因子产生、运输和 Treg 生成的潜在影响，这些是 GVHD 发生的关键因素。为了实现这些目标，我们的团队拥有免疫学、骨髓移植和药物治疗方面的专业知识。独特的转基因、基因敲除和基因敲入小鼠可用于所有计划的实验。拟议的研究肯定会评估 TCN 在临床相关 BMT 模型中控制 GVHD 的功效和机制，并可能为临床预防 GVHD 提供一种新颖且更具选择性的策略。