Exercise as an Immune Adjuvant for Gamma Delta T-cell Therapies in Hematologic Malignancies

运动作为血液恶性肿瘤 Gamma Delta T 细胞疗法的免疫佐剂

• 批准号: 10577605
• 负责人: EMMANUEL KATSANIS
• 金额: $ 63.06万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10577605
• 关键词: Acute Lymphocytic Leukemia; Adjuvant; Adrenergic Agonists; Adrenergic Antagonists; Adrenergic Receptor; Agonist; Allogenic; Antigen Targeting; Antitumor Response; Biological; Blood; CAR T cell therapy; CD19 Antigens; CD19 gene; Cancer Patient; Cells; Cellular immunotherapy; Clinical Trials; Collecting Cell; Cross-Over Trials; Cyclic AMP; Cyclic AMP Receptors; Development; Diagnosis; Exercise; Exhibits; Flow Cytometry; Future; Gene Expression Profile; Genes; Genetic Transcription; Goals; Growth; Hematologic Neoplasms; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Homing; Human; Immunologic Adjuvants; In Vitro; In complete remission; Inferior; Infusion procedures; Interleukin-15; Interleukin-2; Isoproterenol; K-562; K562 Cells; Knock-in; Ligands; Lymphocyte; Membrane Proteins; Methods; Modeling; Mus; Nadolol; Non-Hodgkin' s Lymphoma; Oxidative Phosphorylation; Pathway interactions; Patients; Persons; Phenotype; Placebo Control; Population Heterogeneity; Proliferating; Randomized; Receptor Activation; Recurrent disease; Refractory Disease; Relapse; Rest; Signal Pathway; Signal Transduction; T cell therapy; T-Cell Receptor; T-Lymphocyte; TNFSF10 gene; Therapeutic; Tissues; Treatment Side Effects; Tumor Immunity; Up-Regulation; Variant; Vision; Zoledronate; antagonist; antigen test; arm; bisphosphonate; cancer cell; chimeric antigen receptor; chimeric antigen receptor T cells; cytokine release syndrome; cytotoxic; cytotoxicity; early phase clinical trial; efficacy testing; engineered T cells; experience; graft vs host disease; head-to-head comparison; improved; in vivo; leukemia; leukemia relapse; leukemia/lymphoma; manufacture; metabolic fitness; migration; mouse model; novel; peripheral blood; pre-clinical; preclinical study; programs; protein expression; receptor; response; single-cell RNA sequencing; transcriptome; transcriptomics; tumor; γδ T cells

SUMMARY: Exercise as an Immune Adjuvant for gd T-cell Therapies in Hematologic Malignancies gd T-cells are being considered as an alternative to standard CAR ab T-cells for treating leukemic relapse after hematopoietic stem cell transplantation (HSCT), largely due to their ability to function across MHC barriers without causing graft-versus-host disease (GvHD)1. gd T-cells can be readily expanded in vitro and in vivo using zoledronate (ZOL) and have demonstrated anti-tumor activity in preclinical and early phase clinical trials, but their efficacy against CD19-expressing tumors including acute lymphoblastic leukemia (ALL) and non-Hodgkin’s lymphoma (NHL) has been modest3. Recently, CD19 CAR gd T-cells were found to have profound effects against CD19+ tumors in vitro and in xenogeneic mice, albeit inferior to CD19 CAR ab T-cells, although CD19 CAR gd T-cells were more effective at eliminating CD19 negative escape variants5, 6. As such, if the natural cytotoxicity of gd T-cells could be enhanced they would become a highly attractive “off the shelf” therapeutic option for ALL and NHL. Our goal is to improve gd T-cell therapeutics by collecting “superior” gd T-cells that have been mobilized to peripheral blood by exercise or a synthetic b2-adrenergic receptor (AR) agonist and arming them with a CAR. We will build on several novel and important observations we have made: (i) a single exercise bout instantaneously mobilizes gd T-cells bearing a cytotoxic, co-stimulatory and tissue migration phenotype, allowing their ex vivo manufacture with ZOL+IL-2 to increase by 100-300%4; (ii) exercise expanded gd T-cells have higher in vitro cytotoxicity against several hematologic tumors4 and are more capable of inhibiting K562 leukemic growth in xenogeneic mice, particularly when combined with ZOL sensitization; (iii) exercise skews expanded gd T-cells toward an activated phenotype with heightened NKG2D, TRAIL, DNAM-1 and lowered NKG2A expression, and blocking these activating receptors, or their ligands on K562 cells, abrogates the exercise effects on gd T-cell cytotoxicity; and (iv) the mobilization of these superior gd T-cells with exercise is driven by b2-AR activation4. We hypothesize that exercise will also enhance the quality of CAR gd T-cells by mobilizing gd T-cells with sustained activation of cytotoxicity, co-stimulation, oxidative phosphorylation, homing and proliferation related genes, and that this mobilization will be precipitated by increased cAMP signaling. Our aims are: 1) Determine if a single exercise bout can improve the quality of CAR gd T-cells expanded from healthy donors. 2) Explore the transcriptomic basis for the enhanced expansion and cytotoxicity of exercise mobilized gd T-cells and expanded products. 3) Identify the b2-AR signaling pathways responsible for mobilizing gd T-cells with enhanced expansion and cytotoxicity potential. Our approach involves the use flow cytometry, xenogeneic mouse models, single cell RNA sequencing, and comparisons with CD19 CAR ab T-cells in human trials involving exercise with b-blockers and b-agonist infusion models. We expect these aims to identify underpinning mechanisms and pave the way for a clinical trial whereby exercise/b-agonist mobilized gd T-cells can be collected from donors and cancer patients to increase the potency of CAR T-cell therapies to treat refractory disease and relapse after HSCT.

总结：运动作为血液肿瘤gd T细胞治疗的免疫佐剂 gd T细胞被认为是标准CAR ab T细胞的替代品，用于治疗白血病复发。 造血干细胞移植（HSCT），主要是由于它们能够跨越MHC屏障发挥作用 而不引起移植物抗宿主病（GvHD）1. gd T细胞可以容易地在体外和体内扩增， 唑来膦酸盐（ZOL），并已在临床前和早期临床试验中显示出抗肿瘤活性，但 它们对表达CD 19的肿瘤的疗效，包括急性淋巴细胞白血病（ALL）和非霍奇金淋巴瘤（NHL）。 淋巴瘤（NHL）一直是温和的3。最近，CD 19 CAR gd T细胞被发现对免疫缺陷病毒具有深远的影响。 体外和异种小鼠中的CD 19+肿瘤，尽管不如CD 19 CAR ab T细胞，但CD 19 CAR gd T细胞在消除CD 19阴性逃逸变体方面更有效5，6。因此，如果天然细胞毒性 的gd T细胞可以被增强，它们将成为ALL的一种非常有吸引力的“现成”治疗选择。 还有NHL。我们的目标是通过收集动员的“上级”gd T细胞来改善gd T细胞治疗 通过运动或合成的β 2-肾上腺素能受体（AR）激动剂将它们输送到外周血，并用CAR武装它们。 我们将建立在几个新的和重要的意见，我们已经取得：（一）一个单一的演习回合 瞬时动员携带细胞毒性、共刺激和组织迁移表型的gd T细胞， 它们用ZOL+IL-2的离体生产增加100-300%4;（ii）运动扩增的gd T细胞具有更高的 对几种血液肿瘤的体外细胞毒性4，更能抑制K562白血病生长 在异种小鼠中，特别是当与ZOL致敏组合时;（iii）运动使扩增的gd T细胞偏斜 朝向NKG 2D、TRAIL、DNAM-1表达升高和NKG 2A表达降低的活化表型，和 阻断K562细胞上这些活化受体或其配体，可消除运动对gd T细胞的影响。 细胞毒性;和（iv）这些上级gd T细胞的运动动员是由β 2-AR激活驱动的4。我们 假设运动也将通过持续动员gd T细胞来提高CAR gd T细胞的质量， 细胞毒性、共刺激、氧化磷酸化、归巢和增殖相关基因的活化，以及 这种动员将通过增加的cAMP信号传导而加速。我们的目标是：1）确定是否一个单一的 锻炼可以改善从健康供体扩增的CAR gd T细胞的质量。2)探索 运动动员的gd T细胞的增强扩增和细胞毒性的转录组学基础， 产品. 3)鉴定负责动员增强扩增的gd T细胞的b2-AR信号通路 和细胞毒性潜力。我们的方法包括使用流式细胞术，异种小鼠模型，单细胞 RNA测序，以及在涉及使用b受体阻滞剂运动的人体试验中与CD 19 CAR ab T细胞的比较 和β-激动剂输注模型。我们期望这些目标能够确定基础机制并铺平道路 用于临床试验，由此可以从供体和癌症患者收集运动/β-激动剂动员的gd T细胞。 增加CAR T细胞疗法治疗HSCT后难治性疾病和复发的效力。