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 诊断为晚期上皮性卵巢癌（EOC）的女性中位5年生存率约为20%， 而复发性EOC基本上仍然无法治愈。新型免疫疗法，包括免疫检查点 阻断和嵌合抗原受体（CAR）T细胞疗法已经取得了一些成功，但反应率 通常低于20%。同样，分子靶向治疗的反应率也是 有限，部分原因是缺乏反复突变的靶点和高水平的染色体不稳定性。 EOC的特点。通过PARP抑制和VEGF阻断的合成致死性已经证明了一些 这些新疗法的好处，但耐药机制已经浮出水面。的长期目标 我们的研究计划是开发高效的免疫细胞疗法， 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 高功能性自然杀伤（NK）细胞用于治疗晚期EOC。我们专注于NK细胞，因为i）它们 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，我们建议对人类进行两个根本性的改变， 从健康供体获得的NK细胞。第一个基因改变去除了一个细胞内信号蛋白 使用我们开发的改良CRISPR/Cas9方案抑制NK激活（CISH基因）， 在原代人类NK细胞中非常有效。第二个基因改变是位点特异性整合一个 嵌合抗原受体（CAR）基因，我们已经开发了专门用于NK细胞。这款CAR NK靶向 间皮素蛋白，一种普遍的EOC肿瘤抗原。我们的核心假设是，基因编辑可以 用于克服信号传导抑制并改善NK细胞靶向、持久性和功能， 改善晚期EOC女性患者的治疗结果。本项目拟开展的工作包括 通过与卵巢癌细胞共培养产生并测试基因编辑的NK细胞的增强功能 在体外和体内在细胞系和患者来源的EOC异种移植模型中进行测试。这些数据 临床前试验将用于指导使用这些基因编辑的NK细胞的I期临床试验的设计。 该项目将确定人类基因编辑的CAR NK细胞免疫疗法是否安全并且具有 在复发性EOC的情况下改善结局的潜力。此外，我们从这项研究中学到的东西将 具有更广泛的潜在未来应用于其他恶性肿瘤的影响，因为工程技术 可以快速适应去除不同的负调节因子，并靶向其他肿瘤抗原， 特定的汽车