Gene Edited and Engineered Tumor Cell Therapeutics for Cancer

基因编辑和工程化肿瘤细胞治疗癌症

基本信息

批准号：
10589097
负责人：
Khalid A Shah
金额：
$ 44.23万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-15 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10589097
关键词：
Active immunity Animals Autologous Back Behavior Boston Brain Neoplasms CD8-Positive T-Lymphocytes CRISPR/Cas technology Cell Line Cell Therapy Cells Circulation Clinical Collaborations Cytotoxic agent Data Encapsulated Engineered Gene Engineering Ensure Excision Exhibits Extracellular Matrix Genes Genetic Engineering Glioblastoma Glioma Goals Home Homing Behavior Human Immune Immunity Immunization Immunocompetent Immunomodulators Immunosuppression Immunotherapy Interferon-beta Knock-out Ligands Macrophage Malignant Neoplasms Malignant neoplasm of brain Metastatic/Recurrent Modality Modeling Mus Natural Immunity Neoplasm Metastasis Patients Positron-Emission Tomography Pre-Clinical Model Primary Neoplasm Proteins Publishing Recurrence Recurrent tumor Resected Residual Neoplasm Residual state Resistance Safety Science Site Solid Neoplasm Surgically-Created Resection Cavity Testing Therapeutic Translating Treatment Efficacy Tumor Immunity Tumor Stem Cells adaptive immunity anti-tumor immune response cancer cell cancer therapy cancer type cell killing clinical translation cytotoxic design efficacy evaluation evidence base granulocyte humanized mouse imaging agent imaging biomarker immune modulating agents immunoregulation in vivo mouse model neoplastic cell novel therapeutic intervention novel therapeutics patient prognosis pre-clinical prevent receptor recruit stem cells temozolomide therapeutic genome editing tumor tumor growth tumor microenvironment vaccination strategy

项目摘要

SUMMARY Despite recent advances in therapeutic strategies, the prognosis for patients with highly malignant brain tumors, glioblastomas (GBM) remains poor, with a median survival of 12-19 months. Immunotherapy has emerged as a promising approach for different cancer types. However, its efficacy in GBM has been limited primarily by overall systemic immune suppression and the immune-suppressive tumor micro-environment. Recently, we have shown CRISPR/Cas9 engineered self-targeting re-purposed cancer cells specifically home to tumor cells and release targeted ligands that induce tumor cell killing which translates into survival benefits in mouse models of primary and metastatic tumors. Based on our exciting studies, we have gene edited and subsequently engineered syngeneic immunosuppressive GBM to express bi-functional immunomodulatory and cytotoxic protein, interferon (IFN)β and granulocyte macrophage stimulating factor (GMCSF), which is known to induce both innate and adaptive immunity. Our preliminary data reveal that repurposed immunosuppressive GBM cells do not proliferate in vivo and elicit an active immunity which prevents tumor recurrence. These results although promising, have raised fundamental questions for our tumor cell based gene edited therapy strategy to be characterized and tested extensively in immunocompetent mouse tumor models that mimic clinical settings of immunosuppressive, resected and recurrent immune-profiled GBM tumors. In this proposal, we will first develop and extensively characterize a platform of gene edited and engineered syngeneic immunosuppressive and active GBM therapeutic tumor cells (ThTC) and assess them for their mechanism based direct killing of parental GBM cells and their ability to elicit active anti-tumor immunity in primary and recurrent mouse GBMs. Based on our previous findings that GBM tumor resection promotes the recruitment of CD4/CD8 T cells and local delivery of synthetic extracellular matrix (sECM) encapsulated immunomodulators has therapeutic efficacy, we will test sECM-ThTC for their therapeutic efficacy in resected GBM mouse tumor models. We hypothesize that ThTC will lead to specific killing of residual GBM cells in the tumor resection cavity of primary and recurrent GBMs and elicit active immunity. To ease clinical translation, we will ultimately CRISPR/Cas9 gene edit and subsequently engineer patient derived resected primary tumor cells (hTC) to express human IFN and GMCSF (hThTC). These hThTC will be tested in recurrent GBM models generated from glioma stem cell (GSC) lines in humanized mice. The integration of the safety kill switch, HSV-TK in ThTC will ensure safety in our approach and the incorporation of genetically engineered imaging markers into both ThTC and GBMs will allow us to follow fate and efficacy in vivo and thus to fine tune the proposed approaches. We anticipate that our findings will have a major contribution towards developing novel ThTC based therapies for GBM and are likely to define a new treatment paradigm for patients with other cancers.

概括尽管治疗策略最近取得了进展，但高度恶性脑肿瘤患者的预后，胶质母细胞瘤 (GBM) 的治疗效果仍然较差，免疫疗法的中位生存期为 12-19 个月。然而，它对 GBM 的疗效主要受到整体的限制。最近，我们已经证明了免疫抑制和免疫抑制肿瘤微环境。 CRISPR/Cas9 工程改造自靶向重新定位癌细胞，专门针对肿瘤细胞并释放诱导肿瘤细胞杀伤的靶向配体，这转化为原发性肿瘤小鼠模型的生存获益基于我们令人兴奋的研究，我们进行了基因编辑并随后进行了工程改造。同基因免疫抑制 GBM 表达双功能免疫调节和细胞毒性蛋白、干扰素 (IFN)β 和粒细胞巨噬细胞刺激因子 (GMCSF)，已知可诱导先天性和我们的初步数据表明，重新调整用途的免疫抑制 GBM 细胞不会增殖。这些结果很有希望，并引起积极的免疫，从而防止肿瘤复发。提出了我们基于肿瘤细胞的基因编辑治疗策略的基本问题，以进行表征和主要在模拟免疫抑制临床环境的免疫活性小鼠肿瘤模型中进行测试，在本提案中，我们将首先开发并主要开发切除的和复发的免疫特征GBM肿瘤。表征基因编辑和工程同基因免疫抑制和活性 GBM 的平台治疗性肿瘤细胞 (ThTC) 并评估它们直接杀死亲本 GBM 细胞的机制以及它们在原发性和复发性小鼠 GBM 中引发主动抗肿瘤免疫的能力。研究发现，GBM 肿瘤切除促进了 CD4/CD8 T 细胞的募集和合成的局部递送细胞外基质（sECM）封装的免疫调节剂具有治疗功效，我们将测试sECM-ThTC 我们研究 ThTC 在切除 GBM 小鼠肿瘤模型中的治疗效果。特异性杀死原发性和复发性 GBM 肿瘤切除腔内残留的 GBM 细胞，并引发活性为了便于临床转化，我们最终将进行 CRISPR/Cas9 基因编辑和后续工程。患者来源的切除的原发性肿瘤细胞 (hTC) 表达人 IFN 和 GMCSF (hThTC)。将在人源化小鼠的神经胶质瘤干细胞（GSC）系生成的复发性 GBM 模型中进行测试。在 ThTC 中集成安全终止开关 HSV-TK 将确保我们方法的安全性以及 ThTC 和 GBM 中的基因工程成像标记将使我们能够追踪体内的命运和功效从而对所提出的方法进行微调，我们预计我们的发现将做出重大贡献。开发基于 ThTC 的 GBM 新型疗法，并可能为 GBM 定义新的治疗范式患有其他癌症的患者。