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

基本信息

批准号：
10705051
负责人：
Branden S Moriarity
金额：
$ 23.24万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10705051
关键词：
Activated Natural Killer Cell Address Adult Affect Allogenic Animal Model Antigen Targeting Antigens Ascites B lymphoid malignancy Binding 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 Correlative Study Cytokine Signaling Data Development Diagnosis Disadvantaged Dose Electroporation Engineering Epithelial ovarian cancer Excision Future Genes Genetic Genetic Engineering Goals Guide RNA Human Immune Immunotherapy In Vitro 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 Cell Immunotherapy Natural Killer Cells Neoplasm Metastasis PARP inhibition Patients Peritoneal Phase I Clinical Trials Phase I/II Trial Physiological Preclinical Testing Process Proliferating 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 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 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 site-specific integration success therapy outcome tumor microenvironment tumor progression

项目摘要

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，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，但反应率同样，分子靶向治疗的反应率通常低于 20%。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 的合成致死性通过 PARP 抑制和 VEGF 阻断已得到证实。好处，但耐药机制已经浮出水面，这些新疗法的长期目标。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) 它们确实如此。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项根本性的改变。从健康捐赠者获得的 NK 细胞，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）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。我们专门针对 NK 细胞开发了嵌合抗原受体 (CAR) 基因。间皮素蛋白是一种常见的 EOC 肿瘤抗原，我们的中心假设是基因编辑可以实现。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提出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试验以及体内 EOC 细胞系和患者异种移植模型的测试。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此外，我们从这项研究中学到的东西将有助于改善复发性 EOC 的结果。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。