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: 10540181
• 负责人: MIGUEL A VILLALONA-CALERO
• 金额: $ 62.07万
• 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/10540181
• 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; Cytolysis; Data; Dependence; Disease; Dose; Epithelial; Gene-Modified; Growth Factor Receptors; HLA Antigens; Human; IL18 gene; Immune; Immune checkpoint inhibitor; Immunotherapy; In Vitro; Interleukin-12; Interleukin-15; Interleukin-18; Interleukin-2; K-562; K562 Cells; Knock-out; Lead; 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; Recurrence; 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; in vivo; lung cancer cell; men; mouse model; myeloid leukemia cell; patient safety; 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.

肺癌（LC）是男性和女性癌症死亡的最常见原因， 据估计，2020年美国将有228，820例新病例和135，720例死亡。近年来 抑制免疫检查点，如程序性细胞死亡-1（PD-1）和程序性细胞死亡配体- 1（PD-L1）已被证明为LC患者提供生存益处。然而，大多数患者 表现出原发性耐药或经历肿瘤复发并死于其疾病。 我们的研究小组已经证明，癌细胞上调自然杀伤（NK）细胞上的PD-L1， 可以靶向恶性肿瘤而不需要嵌合抗原受体或预先抗原暴露， 不需要与受体的人类白细胞抗原匹配以获得潜在活性。PD上调- L1能增强NK细胞的功能。此外，PD-L1抑制剂atezolizumab（AZ）导致 针对缺乏PD-L1表达的髓性白血病的增强的白血病细胞杀伤，以及用 选择性细胞因子（IL-12、IL-15和IL-18）与AZ组合显示出显著的改善， 即使在肿瘤组织中不存在PD-L1表达的情况下也能存活。 我们能够表达可溶性IL-15（sIL-15）标记的截短的上皮生长因子受体， 脐带NK细胞，同时上调NK细胞上的内源性PD-L1表达。这些 与模拟转导的NK细胞相比，转导的NK细胞维持大于30%的抗原特异性肿瘤溶解。 NK细胞，并对A549非小细胞LC（NSCLC）细胞表现出细胞毒性。人A549 NSCLC 随后将细胞注射到非同基因小鼠中， 脐血NK细胞。与用模拟转导的NK细胞或表达NK细胞的NK细胞处理相比， sIL-15（sIL-15-NK），但没有离体活化，增强的CB NK细胞诱导实质性减少， 肿瘤体积我们还对这种方法进行了体内安全性/毒性研究，并与单独使用AZ进行了比较。 我们的数据表明，抗PD-L1 mAb治疗在治疗PD-L1阴性患者中具有独特的治疗作用。 癌症，通过PD-L1（+）NK细胞发挥作用。这种活性独立于PD-1活性实现，并且在 存在NK活化细胞因子。我们假设细胞因子“增强”的NK细胞将提供临床应用 这种方法的抗肿瘤活性将进一步增强， 管理AZ。为了验证这一假设并证明这一策略的安全性，我们提出了额外的建议。 体内安全性和有效性研究，随后是1期研究，其中CB NK细胞经遗传修饰， 表达sIL-15，然后在IL-2、IL-18和IL-12存在下进行离体扩增 在淋巴细胞耗竭后，单独或与AZ联合治疗NSCLC患者， 既往在PD-1/PD-L1抑制剂治疗期间或治疗后发生疾病进展。