LTBR CARs as next-generation therapies for R/R lymphoma

LTBR CAR 作为 R/R 淋巴瘤的下一代疗法

• 批准号: 10635791
• 负责人: Catherine Sibyl Diefenbach
• 金额: $ 83.17万
• 依托单位: NEW YORK GENOME CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-10 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10635791
• 关键词: Alanine; Antigens; Antitumor Response; Autologous; B-Cell Activation; B-Cell NonHodgkins Lymphoma; B-Lymphocytes; Biological; Biological Assay; CAR T cell therapy; CD19 gene; CD28 gene; Cancer Burden; Cell Line; Cell physiology; Cell surface; Cells; Clinical; Clinical Trials; Data; Development; Disease; Engineering; FDA approved; Flow Cytometry; Genetic Transcription; Heterogeneity; Histology; Immunologic Memory; Immunology; Immunosuppression; In Vitro; In complete remission; Inflammatory; Intervention; Kinetics; Laboratory Study; Libraries; Lymphocyte; Lymphoma; Measures; Molecular Profiling; Mus; Mutagenesis; Myelogenous; Myeloid Cells; Myeloid-derived suppressor cells; Outcome; Patients; Population; Proteins; RNA; Refractory; Relapse; Research; Resistance; Sampling; Scanning; Signal Transduction; Specificity; Structure of germinal center of lymph node; T cell differentiation; T cell response; T-Cell Activation; T-Cell Lymphoma; T-Lymphocyte; Testing; Toxic effect; Translational Research; Tumor Burden; Variant; Work; Xenograft procedure; activated B cell like; cell killing; chimeric antigen receptor; chimeric antigen receptor T cells; clinical development; clinical efficacy; curative treatments; cytokine; engineered T cells; exhaust; exhaustion; immune resistance; immunological synapse; improved; improved outcome; in vivo; large cell Diffuse non-Hodgkin' s lymphoma; leukemia; lymphotoxin beta receptor; mouse model; multiple omics; next generation; novel; overexpression; peripheral blood; prospective; receptor; relapse patients; response; stemness; tumor; γδ T cells

PROJECT SUMMARY Up to 50% of patients with diffuse large B-cell lymphoma (DLBCL) relapse after first-line treatment. Chimeric antigen receptor (CAR) T-cells have recently emerged as a curative therapy for relapsed or refractory (R/R) DLBCL. However, only 35% of R/R DLBCL patients treated with CAR T-cells have a durable response, and survival is measured in months for patients who fail to benefit. Improvements in CAR T-cells are urgently required to improve outcomes. Recently, we identified the cell surface lymphotoxin beta receptor (LTBR) as a positive T- cell regulator that enhances CD19 CAR T-cell efficacy in vitro and in vivo. LTBR is typically expressed in a subset of myeloid cells but absent in lymphocytes; however when expressed in T-cells, LTBR induces proinflammatory cytokine release, and improves antigen-specific CAR T- and γδ T-cell responses with no appreciable off-target toxicity. Based on these observations, we hypothesize that LTBR can effectively potentiate anti-tumor activity in R/R lymphoma T-cells, reducing markers of T cell exhaustion and outperforming current FDA-approved CAR-Ts across R/R DLBCL subtypes. In Aim 1, we characterize differences in expression of T-cell differentiation, activation, and exhaustion markers and myeloid populations in 25 treatment-naive and 25 R/R DLBCL patient samples. To understand if LTBR can similarly improve CAR-T response in the R/R context, we will use single- cell profiling and functional assays to test autologous CD19+ cell killing, with and without LTBR. In Aim 2 we will evaluate the impact of DLBCL subtype on CAR T-cell activity by introducing LTBR and CAR T-cells into mice xenotransplanted with multiple germinal center B-cell (GCB) and activated B-cell (ABC) cell lines. Since T-cell activation and kinetics are further influenced by patient tumor burden, we will also investigate the efficacy of LTBR-CAR T-cell therapy in a high tumor burden context and test for durable immune memory after complete tumor regression. Recently, by fusing the intracellular signaling domain of LBTR directly to existing (CD28 and 4-1BB) CARs, we have developed a novel CAR construct with more potent antitumor response. In Aim 3 using comprehensive scanning mutagenesis of the LTBR domain, we will create a library of CAR variants and test their ability to improve tumor killing, resistance to exhaustion and cytokine section. We will also measure changes in T-to-B cell immune synapses and resistance to immunosuppression by myeloid-derived suppressor cells (MDSCs) in the most promising LTBR-CARs. This project is the first to comprehensively characterize T cells states in treatment-naive and R/R DLBCL and evaluate LTBR as a T-cell activating strategy to maximize intrinsic anti-tumor activity in R/R DLBCL. There is substantial potential for our work to serve as a bridge from laboratory studies to clinical trials and to help the 40,000 patients per year with R/R DLBCL and other B-cell NHLs.

项目概要 高达 50% 的弥漫性大 B 细胞淋巴瘤 (DLBCL) 患者在一线嵌合治疗后复发。 抗原受体 (CAR) T 细胞最近已成为复发或难治性 (R/R) 的治疗方法 然而，接受 CAR T 细胞治疗的 R/R DLBCL 患者中只有 35% 具有持久缓解，并且 对于未能受益的患者，生存期以月为单位，迫切需要改进 CAR T 细胞。 最近，我们确定了细胞表面淋巴毒素 β 受体 (LTBR) 为阳性 T-受体。 LTBR 是一种增强 CD19 CAR T 细胞体外和体内功效的细胞调节因子，通常在一个亚群中表达。 骨髓细胞中存在，但在淋巴细胞中不存在；然而，当在 T 细胞中表达时，LTBR 会诱导促炎症反应。 细胞因子释放，并改善抗原特异性 CAR T 和 γδ T 细胞反应，且没有明显的脱靶现象 基于这些观察，我们发现 LTBR 可以有效增强抗肿瘤活性。 R/R 淋巴瘤 T 细胞，减少 T 细胞耗竭标志物，性能优于当前 FDA 批准的 CAR-T 在目标 1 中，我们描述了 T 细胞分化表达的差异。 25 名初治患者和 25 名 R/R DLBCL 患者的激活和耗竭标志物以及骨髓细胞群 为了了解 LTBR 是否可以类似地改善 R/R 环境中的 CAR-T 反应，我们将使用单样本。 在目标 2 中，我们将使用和不使用 LTBR 进行细胞分析和功能测定，以测试自体 CD19+ 细胞杀伤作用。 通过将 LTBR 和 CAR T 细胞引入小鼠体内，评估 DLBCL 亚型对 CAR T 细胞活性的影响 异种移植多种生发中心 B 细胞 (GCB) 和活化 B 细胞 (ABC) 细胞系。 激活和动力学进一步受到患者肿瘤负荷的影响，我们还将研究其功效 高肿瘤负荷背景下的 LTBR-CAR T 细胞疗法，并在完成后测试持久免疫记忆 最近，通过将 LBTR 的细胞内信号结构域直接融合到现有的（CD28 和 4-1BB) CAR，我们在 Aim 3 中开发了一种具有更有效抗肿瘤反应的新型 CAR 结构。 LTBR结构域的全面扫描诱变，我们将创建CAR变体库并进行测试 他们提高肿瘤杀伤能力、抗衰竭能力和细胞因子部分的变化。 T-B 细胞免疫突触和髓源性抑制细胞对免疫抑制的抵抗 (MDSC) 在最有前途的 LTBR-CAR 中该项目是第一个全面表征 T 细胞的项目。 治疗初治和 R/R DLBCL 的状态，并评估 LTBR 作为 T 细胞激活策略以最大化内在 我们的工作具有作为实验室桥梁的巨大潜力。 研究到临床试验，每年帮助 40,000 名 R/R DLBCL 和其他 B 细胞 NHL 患者。