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.

根据该项目的第一个目标，工作分为两个领域。第一项研究探索了同种异体环境对癌症T细胞受体定向免疫治疗的调节，并确定了选择性抑制供体浆细胞样树突状细胞中的STAT 1可以降低GVHD的严重程度，同时保留抗肿瘤T细胞应答。这项工作最近刚刚被接受出版血液（Capitini等人）。该项目的第二部分研究了同种异体抗原表达对alloHSCT后抗肿瘤免疫应答抑制的隔室特异性作用，并利用了来源于转基因表达E2 aPBX 1小鼠的前体B细胞白血病系，E2 aPBX 1是一种复发性易位，存在于约5%的儿科ALL中（Bijl et al，Genes and Development，2005），并在我们的实验室中开发成可移植模型。这项工作发表在Biology of Blood and Marrow Transplantation（Shand et al）上。在使用这种可移植ALL的正在进行的工作中，我们已经证明ALL可以通过体内T细胞应答有效靶向，但是这种应答需要接种供体T细胞接种物。其次，同种异体抗原有助于T细胞输注后的治愈，但导致GVHD。最后，来自致敏供体的分选的T细胞群可以介导选择性移植物抗白血病应答。使用这个模型，我们已经开始研究白血病在骨髓中的早期进展以及这种进展对T细胞的影响。我们已经确定，白血病浸润区室中令人惊讶的大百分比的T细胞表达高水平的T细胞功能的负调节因子，程序性死亡1（PD-1）受体。另外的研究表明，PD-1+ T细胞的百分比与白血病累及的程度相关，并且PD-1+ T细胞还表达衰老表型的其他标志物，如T细胞免疫球蛋白和粘蛋白结构域3（Tim-3）。有趣的是，在早期白血病进展期间仔细评估T细胞已经表明，PD 1的诱导发生较早（在注射白血病后第5天），而其他T细胞衰老标志物如Tim-3和淋巴细胞活化基因3（LAG 3）的获得发生较晚，表明这些标志物在抗白血病潜力方面可能更具功能相关性。事实上，来自辐射肿瘤细胞致敏小鼠的T细胞也表达PD 1，但不表达Tim-3或LAG 3，并介导抗白血病作用。最后，来自ALL患者的人骨髓样品白血病样品（从我们的血液学肿瘤生物学研究获得）的初步数据显示了PD 1、Tim-3和LAG 3在T细胞亚群上的表达。总之，这些数据提供了关于ALL在骨髓微环境中的免疫抑制作用如何作为ALL靶向免疫治疗的佐剂被逆转的见解。在该项目的目标2下，我们从James Kochenderfer博士获得了一种鼠CD 19靶向嵌合抗原受体（CAR），与临床环境中使用的受体类似，可在正在进行的临床试验中诱导50-70%的化疗难治性或复发性ALL患者缓解。这些杂合受体（免疫球蛋白抗原结合域和T细胞受体信号传导组分）是在Fry实验室进行的项目ZIA BC 011565的主要重点。使用目标1中描述的CD 19和同种异体移植模型，我们研究了alloHSCT后与CAR治疗相关的免疫生物学。初步数据表明，表达CAR的T细胞可以引起由内源性同种异体反应性TCR介导的GVHD，但这仅在存在CAR抗原时发生，并且这通过CAR T细胞产生细胞因子（主要是IL-6，一种与临床中CD 19 CAR治疗后观察到的主要毒性有关的细胞因子）而加剧。根据该项目的第三个目标，并利用根据ALL对骨髓微环境（T细胞表达耗竭标志物）的影响开发的工具，我们已经开始研究骨髓生态位因子对ALL进展和治疗抗性的作用。我们已经开始基于基因组筛选结果的候选方法，其中发现前B细胞ALL的一个子集过表达胸腺基质基质受体（TSLPR），并且该受体的过表达与较高的复发风险相关。重要的是，TSLP细胞因子结合由TSLPR和IL-7受体α链组成的异二聚体受体。Fry实验室过去在IL-7的免疫生物学方面具有专业知识。使用我们的可移植ALL模型，我们已经证明，通过转导过度表达TSLPR导致白血病的早期进展加速和对化疗药物的耐药性。我们还发现TSLP是由骨髓基质细胞产生的，这表明TSLPR过表达在ALL中的生物学效应是配体依赖性的。我们目前正在探索是否中和TSLP可以逆转TSLPR过表达的影响，这具有潜在的治疗意义。