Project 4 Treatment of Advanced Ovarian Cancer Using Gene-Edited NK CAR Cells

项目4 使用基因编辑的NK CAR细胞治疗晚期卵巢癌

• 批准号: 10452722
• 负责人: Branden S Moriarity
• 金额: $ 24.71万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2026-08-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10452722
• 关键词: Activated Natural Killer Cell; Address; Adult; Affect; Allogenic; Animal Model; Antigen Targeting; Antigens; Ascites; B lymphoid malignancy; Biometry; Blood; CAR T cell therapy; CD19 gene; CISH gene; CRISPR/Cas technology; Cell Line; Cell Survival; Cell Therapy; Cells; Cellular biology; Characteristics; Chromosomal Instability; Chronic; Clinic; Clinical; Clinical Trials; Clinical Trials Design; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Correlative Study; Cytokine Signaling; Data; Development; Diagnosis; Disadvantaged; Dose; Electroporation; Engineering; Epithelial ovarian cancer; Excision; Future; Genes; Genetic; Genetic Engineering; Goals; Guide RNA; Human; Hypersensitivity; Immune; Immunotherapy; Interleukin-15; Intracellular Signaling Proteins; K-562; Knock-in; Knock-out; Knockout Mice; Learning; Ligands; Malignant Neoplasms; Malignant neoplasm of ovary; Maximum Tolerated Dose; Measures; Membrane; Methods; Modification; Mutate; Mutation; NK Cell Activation; NK cell therapy; Natural Killer Cells; Neoplasm Metastasis; PARP inhibition; Patients; Peritoneal; Phase I Clinical Trials; Phase I/II Trial; Physiological; Preclinical Testing; Process; Proteins; Protocols documentation; RNA; Receptor Cell; Receptor Gene; Recombinant adeno-associated virus (rAAV); Recurrence; Refractory; Research; Resistance; Resources; Signal Pathway; Signal Transduction; Site; Solid Neoplasm; Specificity; Surface; Survival Rate; T cell therapy; T-Lymphocyte; Techniques; Testing; Toxic effect; Tumor Antigens; Vascular Endothelial Growth Factors; Viral; Woman; Work; arm; base; cancer cell; chimeric antigen receptor; cytokine; cytokine release syndrome; design; engineered NK cell; exhaustion; graft vs host disease; immune checkpoint blockade; improved; improved outcome; in vitro testing; in vivo; interleukin-21; mesothelin; molecular targeted therapies; novel; novel therapeutics; patient derived xenograft model; peripheral blood; phase 1 designs; programs; resistance mechanism; response; secondary outcome; side effect; success; therapy outcome; tumor microenvironment; tumor progression

ABSTRACT – PROJECT 4 Women diagnosed with advanced epithelial ovarian cancer (EOC) have a median 5-year survival rate of ~20%, and recurrent EOC remains essentially incurable. Novel immunotherapies, including immune checkpoint blockade and chimeric antigen receptor (CAR) T cell therapies have had some success, but response rates have been typically below 20%. Similarly, the response rates to molecularly targeted therapies have been limited, partly due to the lack of recurrently mutated targets and the high level of chromosomal instability characteristic of EOC. Synthetic lethality via PARP inhibition and VEGF blockade have demonstrated some benefit, but mechanisms of resistance have already surfaced for these new therapies. The long-term goal of our research program is to develop highly effective immune cell-based therapies capable of eliminating recurrent EOC. In Project 4, we will use advanced gene editing techniques that we have developed to produce hyperfunctional Natural Killer (NK) cells for treating advanced EOC. We focus on NK cells because i) they do not require antigen priming and ii) side effects, such as graft versus host disease and cytokine release syndrome, are minimal compared to T cell therapy. We propose to make two fundamental alterations to human NK cells obtained from healthy donors. The first genetic alteration removes an intracellular signaling protein that dampens NK activation (the CISH gene) using a modified CRISPR/Cas9 protocol we have developed that is highly effective in primary human NK cells. The second genetic alteration is to site-specifically integrate a chimeric antigen receptor (CAR) gene that we have developed specifically for NK cells. This CAR NK targets the mesothelin protein, a prevalent EOC tumor antigen. Our central hypothesis is that gene editing can be used to overcome signaling inhibition and improve NK cell targeting, persistence and function, resulting in improved therapeutic outcomes for women with advanced EOC. The work proposed in this project includes generating and testing the gene edited NK cells for enhanced function by co-culturing with ovarian cancer cells in vitro and testing in both cell line- and patient-derived xenograft models of EOC in vivo. The data from these preclinical tests will be used to guide the design of the Phase I clinical trial using these gene-edited NK cells. This project will determine whether gene-edited CAR NK cell immunotherapy in humans is safe and has the potential to improve outcomes in the setting of recurrent EOC. In addition, what we learn from this study will have the broader impact of potential future application to other malignancies, as the engineering techniques can be quickly adapted to remove different negative regulators and to target other tumor antigens with NK- specific CARs.

摘要 – 项目 4 被诊断患有晚期上皮性卵巢癌 (EOC) 的女性的中位 5 年生存率约为 20%， 复发性 EOC 基本上仍然无法治愈。新型免疫疗法，包括免疫检查点 封锁和嵌合抗原受体 (CAR) T 细胞疗法取得了一些成功，但反应率 通常低于 20%。同样，分子靶向治疗的反应率 有限，部分原因是缺乏经常突变的靶点和高度的染色体不稳定性 EOC 的特点。通过 PARP 抑制和 VEGF 阻断产生的合成致死率已被证明具有一定的杀伤力 好处，但这些新疗法的耐药机制已经浮出水面。长期目标是 我们的研究计划是开发高效的基于免疫细胞的疗法，能够消除 复发性EOC。在项目4中，我们将使用我们开发的先进基因编辑技术来生产 用于治疗晚期 EOC 的功能亢进的自然杀伤 (NK) 细胞。我们专注于 NK 细胞，因为 i) 它们确实 不需要抗原引发和 ii) 副作用，例如移植物抗宿主病和细胞因子释放 与 T 细胞疗法相比，该综合征的发生率极低。我们建议对人类进行两项根本性的改变 NK 细胞取自健康捐赠者。第一个基因改变去除了细胞内信号蛋白 使用我们开发的改良 CRISPR/Cas9 协议抑制 NK 激活（CISH 基因） 对原代人类 NK 细胞非常有效。第二个基因改变是位点特异性整合 我们专门为NK细胞开发的嵌合抗原受体（CAR）基因。这个CAR NK的目标 间皮素蛋白，一种常见的 EOC 肿瘤抗原。我们的中心假设是基因编辑可以 用于克服信号传导抑制并改善 NK 细胞的靶向性、持久性和功能，从而 改善晚期 EOC 女性的治疗结果。该项目建议的工作包括 通过与卵巢癌细胞共培养来生成和测试基因编辑的 NK 细胞以增强功能 体外和在细胞系和患者来源的 EOC 异种移植模型中进行体内测试。来自这些的数据 临床前测试将用于指导使用这些基因编辑 NK 细胞的 I 期临床试验的设计。 该项目将确定基因编辑的 CAR NK 细胞免疫疗法在人类中是否安全并具有 改善复发性 EOC 的结果的潜力。此外，我们从这项研究中学到的东西将 随着工程技术的发展，未来对其他恶性肿瘤的潜在应用具有更广泛的影响 可以快速适应去除不同的负调节因子并用 NK- 靶向其他肿瘤抗原 特定的 CAR。