ALL immunobiology and the bone marrow niche

ALL 免疫生物学和骨髓生态位

• 批准号: 8938043
• 负责人: Terry Fry
• 金额: $ 37.7万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8938043
• 关键词: Acute Lymphocytic Leukemia; Adjuvant; Alloantigen; Allogeneic Bone Marrow Transplantation; Allogenic; Antigens; Area; B-Cell Acute Lymphoblastic Leukemia; Binding; Biological Preservation; Biology; Blood; Bone Marrow; Bone Marrow Transplantation; CD19 gene; Cancer Etiology; Cell physiology; Cessation of life; Child; Childhood Acute Lymphocytic Leukemia; Chronic Lymphocytic Leukemia; Clinic; Clinical; Clinical Trials; Data; Dendritic Cells; Development; Disease remission; Environment; Genes; Genomics; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Homologous Transplantation; Hybrids; Immune; Immune response; Immune system; Immunobiology; Immunoglobulins; Immunologics; Immunosuppression; Immunotherapeutic agent; Immunotherapy; Incidence; Inferior; Infusion procedures; Injection of therapeutic agent; Interleukin-6; Interleukin-7; Laboratories; Ligands; Lymphocyte Activation; Malignant Childhood Neoplasm; Malignant Neoplasms; Mediating; Modeling; Mucins; Mus; Myeloid Leukemia; Outcome; Patients; Phenotype; Population; Production; Publications; Publishing; Reaction; Receptor Signaling; Recurrence; Refractory; Relapse; Resistance; Risk; Role; STAT1 gene; Sampling; Severities; Sorting - Cell Movement; Stromal Cells; Study models; Supportive care; T cell response; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; TSLP gene; Therapeutic; Tissues; Toxic effect; Transgenic Organisms; Transplantation; Vaccination; Work; antigen binding; base; cancer therapy; chemotherapeutic agent; chemotherapy; chimeric antigen receptor; cytokine; exhaustion; graft vs host disease; graft vs leukemia effect; high risk; human TSLP protein; human data; immunoglobulin receptor; in vivo; insight; leukemia; mortality; neoplastic cell; overexpression; prevent; programs; receptor; response; screening; senescence; tool; tumor

Under the first aim of this project work has been divided into 2 areas. The first explored modulation of cancer T cell receptor directed immunotherapy by the allogeneic environment and identified that selective inhibition of STAT1 in donor plasmacytoid dendritic cells could reduce the severity of GVHD with preservation of anti-tumor T cell responses. This work has just recently been accepted for publication Blood (Capitini et al). The second part of the project studied compartment-specific effects of alloantigen expression on inhibition of antitumor immune responses following alloHSCT and utilized a precursor B cell leukemia line derived from mice with transgenic expression of E2aPBX1, a recurring translocation present in approximately 5% of pediatric ALL (Bijl et al, Genes and Development, 2005) and developed in our laboratory into a transplantable model. This work was published in Biology of Blood and Marrow Transplantation (Shand et al). In ongoing work using this transplantable ALL we have demonstrated that ALL can be effectively targeted by a T cell response in vivo but that this response requires vaccination of the donor T cell inocula. Second, allogeneic antigens contribute to the cure following T cell infusion but results in GVHD. Finally, sorted populations of T cells from primed donors can mediate selective graft versus leukemia responses. Using this model we have begun studying the early progression of the leukemia in bone marrow and the impact of this progression on T cells. We have identified that a surprisingly large percentage of T cells in leukemia-infiltrated compartments express high levels of the negative regulator of T cell function, programmed death 1 (PD-1) receptor. Addition studies have shown that the percentage of PD-1+ T cells correlates with the extent of leukemic involvement and that PD-1+ T cells also express other markers of a senescent phenotype such as T cell immunoglobulin and mucin domain 3 (Tim-3). Interestingly, careful assessment T cells during early leukemia progression have shown that the induction of PD1 occurs early (by day 5 after injection of leukemia) whereas acquisition of other T cells senescent markers such as Tim-3 and Lymphocyte Activation Gene 3 (LAG3) occur later suggesting that these markers may be more functionally relevant in terms of antileukemic potential. Indeed, T cells from irradiated tumor cell primed mice also express PD1 but not Tim-3 or LAG3 and mediate an antileukemic effect. Finally, preliminary a data from human bone marrow samples leukemia samples from patients with ALL (obtained from our Hematologic Malignancy Biology Study) have shown expression of PD1, Tim-3 and LAG3 on a subset of T cells. In summary, this data provides insights into how immunosuppressive effects of ALL in the bone marrow microenvironment may be reversed as an adjuvant to ALL-targeted immunotherapy. Under Aim2 of this project we have obtained a murine CD19-targeted chimeric antigen receptor (CAR) from Dr. James Kochenderfer analogous to those used in the clinical setting to induce remission in 50-70% of patients with chemotherapy-refractory or recurrent ALL in ongoing clinical trials. These hybrid receptors (Immunoglobulin antigen binding domain and T cell receptor signaling components) are the primary focus of project ZIA BC 011565 being conducted in the Fry laboratory. Using the CD19 and the allogeneic transplant models described in Aim 1 we have study the immunobiology associated with CAR therapy after alloHSCT. Preliminary data ahs demonstrated that CAR expressing T cells can cause GVHD mediated by the endogenous alloreactive TCR but that this occurs only when there CAR antigen present and that this is exacerbated by CAR T cell production of cytokines (mainly IL-6, a cytokine implicated in major toxicity observed after CD19 CAR therapy in the clinic). Under the third Aim of this project and utilizing tools developed under Aim where the effect of ALL on the bone marrow microenvironment (T cell expression of exhaustion markers) we have begun studying the role of bone marrow niche factors on the progression and therapeutic resistance of ALL. We have started with a candidate approach based on the results of genomic screening where it was found that a subset of pre B cell ALL overexpresses the thymic stromal stromal receptor (TSLPR) and that overexpression of this receptor is associated with a higher risk of relapse. Importantly, the TSLP cytokine binds to a heterodimeric receptor comprised of TSLPR and the IL-7 receptor alpha chain. The Fry laboratory has past expertise in the immunobiology of IL-7. Using our transplantable ALL model we have demonstrated that overexpression of TSLPR by transduction results in accelerated early progression of leukemia and resistance to chemotherapeutic agents. We have also shown that TSLP is produced by bone marrow stromal cells suggesting that the biologic effect of TSLPR overexpression in ALL is ligand-dependent. We are currently exploring whether neutralization of TSLP can reverse the impact of TSLPR overexpression which has potential therapeutic implications.

在这项项目的第一个目标下，工作已分为两个领域。首次通过同种异体环境对癌症T细胞受体进行了定向免疫疗法的调节，并确定在供体浆细胞类动物树突状细胞中对STAT1的选择性抑制可以降低GVHD的严重程度，并保留抗肿瘤T细胞反应。这项工作刚刚被接受出版血液（Capitini等人）。 The second part of the project studied compartment-specific effects of alloantigen expression on inhibition of antitumor immune responses following alloHSCT and utilized a precursor B cell leukemia line derived from mice with transgenic expression of E2aPBX1, a recurring translocation present in approximately 5% of pediatric ALL (Bijl et al, Genes and Development, 2005) and developed in our laboratory into a transplantable 模型。这项工作发表在血液和骨髓移植生物学上（Shand等）。在正在进行的工作中，我们已经证明，所有人都可以在体内有效地靶向所有人，但是这种反应需要对供体T细胞接种物进行疫苗接种。其次，同种异体抗原在输注T细胞后有助于治愈，但导致GVHD。最后，从启动供体中分类的T细胞种群可以介导选择性移植物与白血病反应。使用该模型，我们已经开始研究骨髓中白血病的早期进展以及该进展对T细胞的影响。我们已经确定，白血病室中的T细胞中的T细胞中有很大比例的T细胞表达了T细胞功能的负调节剂，即编程死亡1（PD-1）受体。添加研究表明，PD-1+ T细胞的百分比与白血病受累程度相关，并且PD-1+ T细胞还表达了衰老表型的其他标记，例如T细胞免疫球蛋白和粘蛋白结构域3（TIM-3）。有趣的是，在白血病早期进展过程中仔细评估的T细胞表明，PD1的诱导发生早期（到注射白血病后的第5天），而获得其他T细胞的衰老标记物（如TIM-3和淋巴细胞激活基因3（LAG3））后来提出这些标记可能在这些标记中可能具有更多功能相关的蚂蚁。实际上，来自辐照肿瘤细胞的T细胞还表达PD1，但不表达TIM-3或LAG3，并介导抗白血病作用。最后，来自人类骨髓样品的初步A数据（从我们的血液系统恶性肿瘤生物学研究中获得）的患者的白血病样品显示了PD1，TIM-3和LAG3在T细胞的一部分上的表达。总而言之，这些数据提供了有关所有在骨髓微环境中如何作为对所有目标免疫疗法的佐剂的辅助性的见解。在该项目的AIM2下，我们从James Kochenderfer博士获得了鼠CD19靶向的嵌合抗原受体（CAR），类似于临床环境中使用的鼠类CD19抗原受体（CAR），以诱导50-70％的化学疗法 - 反应 - 恐怖症或重新出现的患者的缓解。这些杂交受体（免疫球蛋白抗原结合结构域和T细胞受体信号传导成分）是在FRY实验室进行Zia BC 011565的主要重点。使用AIM 1中描述的CD19和同种异体移植模型，我们研究了与AllOHSCT后与CAR治疗相关的免疫生物学。 Preliminary data ahs demonstrated that CAR expressing T cells can cause GVHD mediated by the endogenous alloreactive TCR but that this occurs only when there CAR antigen present and that this is exacerbated by CAR T cell production of cytokines (mainly IL-6, a cytokine implicated in major toxicity observed after CD19 CAR therapy in the clinic).在该项目的第三个目标下，并利用了在所有目标中开发的工具，即所有对骨髓微环境的影响（T细胞表达疲劳标记物的T细胞表达）我们已经开始研究骨髓小裂因子对所有人的进展和治疗性耐药性的作用。我们从基于基因组筛查的结果开始的候选方法开始，发现全部B细胞的一部分过表达了胸腺基质受体（TSLPR），并且该受体的过表达与复发的较高风险有关。重要的是，TSLP细胞因子与由TSLPR和IL-7受体α链组成的异二聚体受体结合。 Fry实验室在IL-7的免疫生物学方面具有过去的专业知识。使用我们的移植所有模型，我们已经证明，通过转导的过表达TSLPR会导致白血病的早期进展和对化学治疗剂的抗性加速。我们还表明，TSLP是由骨髓基质细胞产生的，这表明TSLPR过表达的生物学作用在所有方面都是配体依赖性的。我们目前正在探索TSLP的中和是否可以扭转具有潜在治疗意义的TSLPR过表达的影响。