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

• 批准号: 10386860
• 负责人: Khalid A Shah
• 金额: $ 45.11万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 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/10386860
• 关键词: Active immunity; Animals; Autologous; Back; Behavior; Blood Circulation; Boston; Brain Neoplasms; CD8-Positive T-Lymphocytes; CRISPR/Cas technology; Cell Line; Cell Therapy; Cells; Clinical; Collaborations; Cytotoxic agent; Data; Encapsulated; Engineered Gene; Engineering; Ensure; Excision; Exhibits; Extracellular Matrix; Genes; Genetic Engineering; Glioblastoma; Glioma; Goals; HSV-Tk Gene; Home; Homing Behavior; Human; Immune; Immunity; Immunization; Immunocompetent; Immunomodulators; Immunosuppression; Immunotherapy; Interferon-beta; Interferons; Knock-out; Ligands; Malignant Neoplasms; Malignant neoplasm of brain; Modeling; Mus; Natural Immunity; Neoplasm Metastasis; Patients; Positron-Emission Tomography; Pre-Clinical Model; Primary Neoplasm; Proteins; Publishing; Recurrence; Recurrent tumor; Resected; Residual Tumors; 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; base; cancer cell; cancer therapy; cancer type; cell killing; clinical translation; clinically translatable; cytotoxic; design; efficacy evaluation; evidence base; granulocyte; humanized mouse; imaging agent; imaging biomarker; immunoregulation; in vivo; macrophage; 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

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.

摘要 尽管治疗策略最近取得了进展，但高度恶性脑瘤患者的预后， 胶质母细胞瘤(GBM)仍然很差，中位生存期为12-19个月。免疫疗法已经成为一种 针对不同癌症类型的前景看好的方法。然而，它在GBM中的疗效主要受到总体上的限制。 全身免疫抑制与免疫抑制的肿瘤微环境。最近，我们展示了 CRISPR/Cas9工程自靶向重新定位的癌细胞特异性地定位于肿瘤细胞并释放 靶向配体诱导肿瘤细胞杀伤，从而转化为对小鼠原发肿瘤模型的生存益处 和转移性肿瘤。基于我们令人兴奋的研究，我们对基因进行了编辑，并随后进行了工程设计 同基因免疫抑制基因表达双功能免疫调节和细胞毒蛋白干扰素 干扰素β和粒细胞巨噬细胞刺激因子，已知可诱导先天和 适应性免疫。我们的初步数据显示，改变用途的免疫抑制的GBM细胞不会增殖 在体内，并诱导主动免疫，防止肿瘤复发。这些结果虽然很有希望，但也有 提出了我们的基于肿瘤细胞的基因编辑治疗策略要特色化和 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了广泛的测试， 切除和复发的免疫表型基底膜肿瘤。在这项建议中，我们将首先发展和广泛 基因编辑和工程化同种免疫抑制活性基底膜平台的鉴定 治疗性肿瘤细胞(THTC)及其对亲本GBM细胞的直接杀伤机制 以及它们在原发和复发小鼠肾小球系膜细胞中诱导主动抗肿瘤免疫的能力。基于我们之前的 基底膜肿瘤切除促进CD4/CD8T细胞的募集和局部合成 细胞外基质(SECM)包裹的免疫调节剂具有治疗效果，我们将测试SECM-THTC 在切除的GBM小鼠肿瘤模型中的治疗效果。我们假设THTC将导致 原发和复发基底膜肿瘤切除腔内残留基底膜细胞的特异性杀伤作用及活性 豁免权。为了简化临床翻译，我们最终将对CRISPR/Cas9基因进行编辑并随后进行工程 患者切除的原代肿瘤细胞表达人干扰素和巨噬细胞集落刺激因子(HThTC)。这些hThTC 将在人源化小鼠的胶质瘤干细胞(GSC)系产生的复发GBM模型中进行测试。这个 在THTC中集成安全终止开关HSV-TK将确保我们进场的安全性，并将 THTC和GBM中的基因工程成像标记将使我们能够在体内跟踪命运和疗效 从而对拟议的方法进行微调。我们预计，我们的发现将有重大贡献 为GBM开发基于THTC的新疗法，并可能定义一种新的治疗模式 患有其他癌症的患者。