Phase 1 Evaluation of Enhanced Natural Killer Cells as a Treatment Strategy in Non-Small cell Lung Cancer Patients Refractory to PD-1/PD-L1 Immune Checkpoint Inhibitors

增强型自然杀伤细胞作为对 PD-1/PD-L1 免疫检查点抑制剂耐药的非小细胞肺癌患者的治疗策略的 1 期评估

• 批准号: 10680537
• 负责人: MIGUEL A VILLALONA-CALERO
• 金额: $ 61.26万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-09 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10680537
• 关键词: A549; Acute Myelocytic Leukemia; Aftercare; Allogenic; Animals; Antigens; Biological; Blood; Cancer Etiology; Cancer Patient; Cell Line; Cell physiology; Cells; Cessation of life; Clinical; Clinical Trials; Combined Modality Therapy; Cytolysis; Data; Dependence; Disease; Dose; Engraftment; Epithelium; Gene Modified; Growth Factor Receptors; HLA Antigens; Human; IL18 gene; Immune; Immune checkpoint inhibitor; Immunotherapy; In Vitro; Interleukin-12; Interleukin-15; Interleukin-2; K-562; K562 Cells; Knock-out; Leukemic Cell; Lymphocyte Depletion; Malignant Neoplasms; Malignant neoplasm of lung; Messenger RNA; Monoclonal Antibody Therapy; Mus; Myeloid Leukemia; NK Cell Activation; Natural Killer Cells; Non-Small-Cell Lung Carcinoma; PDL1 inhibitors; Patients; Placebos; Proteins; Proto-Oncogene Proteins c-akt; Recurrent tumor; Refractory; Resistance; Role; Safety; Signal Transduction; Testing; Therapeutic; Time; Toxic effect; Tumor Antigens; Tumor Burden; Tumor Tissue; Tumor Volume; Umbilical Cord Blood; Umbilical cord structure; United States; Up-Regulation; Woman; anti-PD-L1; antibody-dependent cell cytotoxicity; cancer cell; cell killing; checkpoint inhibition; chimeric antigen receptor; cytokine; cytotoxicity; efficacy evaluation; efficacy study; experience; first-in-human; improved; in vivo; lung cancer cell; men; mouse model; myeloid leukemia cell; peripheral blood; phase 1 study; phase 1 testing; programmed cell death ligand 1; programmed cell death protein 1; retroviral transduction; safety and feasibility; safety study; treatment strategy; tumor; tumor progression

PROJECT SUMMARY: Lung cancer (LC), the most frequent cause of cancer deaths for men and women, is estimated to lead to 228,820 new cases and 135,720 deaths in the United States in 2020. In recent years inhibition of immune checkpoints, such as programmed cell death-1 (PD-1) and programmed cell death ligand- 1 (PD-L1), has been shown to provide survival benefits to patients with LC. However, most patients demonstrate either primary resistance or experience tumor recurrence and die of their disease. Our group has demonstrated that cancer cells upregulate PD-L1 on natural killer (NK) cells, immune cells that can target malignancies without the necessity of chimeric antigen receptors or prior antigen exposure and do not require matching to recipient's human leukocyte antigen for potential activity. Upregulation of PD- L1resulted in enhanced NK-cell function. Furthermore, the PD-L1 inhibitor atezolizumab (AZ) resulted in enhanced leukemic cell killing against myeloid leukemia lacking PD-L1 expression, and mice treated with selective cytokines (IL-12, IL-15, and IL-18) in combination with AZ showed a significant improvement in survival even in the absence of PD-L1 expression in their tumor tissue. We were able to express soluble IL-15 (sIL-15) tagged with a truncated epithelial growth factor receptor in umbilical cord NK cells in vitro, while upregulating endogenous PD-L1 expression on the NK cells. These transduced NK cells maintained greater than 30% antigen-specific tumor lysis compared to mock-transduced NK cells and demonstrated cytotoxicity against A549 Non-Small-Cell LC (NSCLC) cells. Human A549 NSCLC cells were subsequently injected in non-syngeneic mice and followed with treatment with these “enhanced” cordon blood CB NK cells. In comparison to treatment with mock-transduced NK cells, or NK cells expressing sIL-15 (sIL-15-NK) but without ex vivo activation, the enhanced CB NK cells induced substantial reduction in tumor volume. We also performed safety/toxicity in vivo studies of this approach and compared with AZ alone. Our data suggest that anti-PD-L1 mAb therapy has a unique therapeutic role in treating PD-L1 negative cancer, acting through PD-L1(+) NK cells. This activity is achieved independent of PD-1 activity and in the presence of NK-activating cytokines. We hypothesize that cytokine “enhanced” NK cells will provide clinical benefit to NSCLC patients and that the antitumor activity of this approach will be further enhanced by co- administration of AZ. To test this hypothesis and document the safety of this strategy, we propose additional in vivo safety and efficacy studies followed by a phase 1 study in which CB NK cells, genetically modified to express sIL-15, followed by ex-vivo expansion in the presence of IL-2, IL-18, and IL-12 will be administered either by themselves or combined with AZ, following lymphocyte depletion, to NSCLC patients whose tumor has previously progressed on or after treatment with PD-1/PD-L1 inhibitors.

项目摘要：肺癌 (LC) 是导致男性和女性癌症死亡的最常见原因。 预计 2020 年美国将出现 228,820 例新病例和 135,720 例死亡。 抑制免疫检查点，例如程序性细胞死亡-1 (PD-1) 和程序性细胞死亡配体- 1 (PD-L1) 已被证明可为 LC 患者提供生存益处，但是，大多数患者。 要么表现出原发性耐药，要么经历肿瘤复发和疾病死亡。 我们的团队已经证明癌细胞上调自然杀伤 (NK) 细胞（免疫细胞）上的 PD-L1 无需嵌合抗原受体或事先暴露抗原即可靶向恶性肿瘤 不需要与受体的人类白细胞抗原匹配即可上调 PD-的潜在活性。 L1 导致 NK 细胞功能增强，此外，PD-L1 抑制剂 atezolizumab (AZ) 导致 NK 细胞功能增强。 增强对缺乏 PD-L1 表达的骨髓性白血病的白血病细胞杀伤作用，以及用 选择性细胞因子（IL-12、IL-15 和 IL-18）与 AZ 联合使用可显着改善 即使肿瘤组织中没有 PD-L1 表达，患者也能存活。 我们能够在细胞中表达带有截短上皮生长因子受体标记的可溶性 IL-15 (sIL-15) 体外研究脐带 NK 细胞，同时上调 NK 细胞上内源性 PD-L1 的表达。 与模拟转导的相比，转导的 NK 细胞保持了超过 30% 的抗原特异性肿瘤溶解 NK 细胞并对 A549 非小细胞 LC (NSCLC) 细胞具有细胞毒性。 随后将细胞注射到非同基因小鼠体内，并用这些“增强型”进行治疗 Cordon 血 CB NK 细胞与模拟转导的 NK 细胞或表达 NK 细胞的治疗相比。 sIL-15 (sIL-15-NK) 但没有离体激活，增强的 CB NK 细胞诱导 我们还对这种方法进行了安全性/毒性体内研究，并与单独的 AZ 进行了比较。 我们的数据表明，抗 PD-L1 mAb 疗法在治疗 PD-L1 阴性方面具有独特的治疗作用 癌症，通过 PD-L1(+) NK 细胞发挥作用，这种活性的实现与 PD-1 活性无关。 我们认为细胞因子“增强”的 NK 细胞将提供临床作用。 对 NSCLC 患者有益，并且该方法的抗肿瘤活性将通过联合进一步增强 为了检验这一假设并记录该策略的安全性，我们提出了额外的建议。 体内安全性和有效性研究随后进行了第一阶段研究，其中 CB NK 细胞经过基因改造 表达 sIL-15，然后在 IL-2、IL-18 和 IL-12 存在的情况下进行离体扩增 单独使用或在淋巴细胞清除后与 AZ 联合治疗患有肿瘤的非小细胞肺癌 (NSCLC) 患者 既往在使用 PD-1/PD-L1 抑制剂治疗时或治疗后出现进展。