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.

总结 尽管最近在治疗策略方面取得了进展，但高度恶性脑肿瘤患者的预后， 胶质母细胞瘤（GBM）仍然很差，中位生存期为12-19个月。免疫疗法已经成为一种 有希望的方法来治疗不同类型的癌症。然而，其在GBM中的疗效主要受到总体 系统性免疫抑制和免疫抑制性肿瘤微环境。最近，我们发现 CRISPR/Cas9工程化的自我靶向重新利用的癌细胞特异性地回到肿瘤细胞并释放 靶向配体诱导肿瘤细胞杀伤，这转化为原发性肝癌小鼠模型中的生存益处， 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， 表达双功能免疫调节和细胞毒性蛋白干扰素同基因免疫抑制性GBM (IFN)β和粒细胞巨噬细胞刺激因子（GMCSF），已知其诱导先天性和 适应性免疫我们的初步数据显示，重新利用的免疫抑制GBM细胞不增殖， 并引发防止肿瘤复发主动免疫。这些结果虽然令人鼓舞， 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， 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测试， 切除和复发的免疫特征GBM肿瘤。在这一建议中，我们将首先发展和广泛地 表征基因编辑和工程化的同基因免疫抑制和活性GBM的平台 治疗性肿瘤细胞（ThTC），并评估它们基于直接杀死亲本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。基于我们之前 GBM肿瘤切除术促进了CD 4/CD 8 T细胞的募集和合成T细胞的局部递送。 细胞外基质（sECM）包裹的免疫调节剂具有治疗功效，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会导致 特异性杀伤原发性和复发性GBM的肿瘤切除腔中的残留GBM细胞，并引发活性 免疫力为了简化临床翻译，我们最终将对CRISPR/Cas9基因进行编辑， 患者来源的切除的原代肿瘤细胞（hTC）以表达人IFN γ和GMCSF（hThTC）。这些hThTC 将在人源化小鼠中由神经胶质瘤干细胞（GSC）系产生的复发性GBM模型中进行测试。的 在ThTC中集成安全终止开关HSV-TK将确保我们的方法的安全性， 将基因工程成像标记物植入ThTC和GBM中将使我们能够在体内跟踪命运和功效 从而微调所提出的方法。我们预计我们的发现将对 为GBM开发基于ThTC的新疗法，并可能为GBM定义新的治疗模式。 其他癌症患者。