Neural Stem Cell Carriers for Glioblastoma Immunotherapy

用于胶质母细胞瘤免疫治疗的神经干细胞载体

• 批准号: 9297711
• 负责人: Irina V Balyasnikova
• 金额: $ 38.89万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9297711
• 关键词: Adenoviruses; Adoptive Transfer; Adult; Affinity; Aftercare; Allogenic; Alpha Cell; Animals; Antibodies; Antigens; Autoimmune Process; Autologous; Beds; Brain; Brain Neoplasms; CD3 Antigens; Cell Communication; Cell Line; Cell Surface Receptors; Cells; Clinical; Clinical Trials; Continuous Infusion; Coupled; Development; Diagnosis; Engineering; Ensure; Expectancy; FDA approved; Gene Expression; Gene Expression Profile; Generations; Genetic; Glioblastoma; Glioma; Goals; Human; Hybridomas; IL2 gene; Immune; Immune response; Immunotherapy; In Vitro; Kidney; Legal patent; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Mesenchymal; Modality; Modeling; Modification; Monoclonal Antibodies; Oncolytic; Patient-Focused Outcomes; Patients; Peripheral; Pharmaceutical Preparations; Phase; Phase I Clinical Trials; Pre-Clinical Model; Production; Proliferating; Recombinant Proteins; Recurrence; Research; Risk; Site; Specificity; T cell therapy; T-Cell Activation; T-Lymphocyte; TNF gene; Testing; Therapeutic; Therapeutic Agents; Therapeutic Effect; Tissues; Toxic effect; Translations; Tropism; Tumor Antigens; Xenograft Model; Xenograft procedure; brain cell; cancer cell; cancer therapy; cellular engineering; clinical application; cost; cytokine; cytotoxic; experimental study; improved; in vivo; killings; mouse model; neoplasm immunotherapy; neoplastic cell; nerve stem cell; novel therapeutics; outcome forecast; overexpression; pre-clinical; preclinical study; response; safety testing; selective expression; suicide gene; tumor; tumor progression

Glioblastoma (GBM) is the most aggressive malignant brain cancer in adults. People diagnosed with GBM have limited therapeutic options and short survival expectancies. A major problem with existing therapeutic approaches is their lack of specificity for neoplastic cells, which results in substantial treatment toxicity. Antibody-mediated specific targeting of tumor-associated antigens has been a successful strategy for cancer therapy as it limits the off-target effect of systemically infused drugs. Genetic modifications of such antibodies coupled with efficient delivery strategies can greatly improve the anti-tumor efficacy of these molecules. One such modification is bi-specific tandem single–chain antibodies (biscFv) that promote T-cell- tumor cell interactions that, in turn, kill the tumor cells. However, biscFv have short half-lives and fast clearance, necessitating frequent or continuous infusions to achieve therapeutic effect. We propose to overcome these hurdles through the generation of neural stem cells (NSCs) producing biscFv. NSCs are able to track brain tumor cells after systemic, local, and intranasal delivery, and efficiently deliver therapeutic payload to tumors sites in preclinical models of GBM. NSCs secreting biscFv can be directly mixed with autologous patients T cells for the production of a local immune response aimed at eradicating tumors. Recently, we developed and characterized a monoclonal antibody specifically targeting IL13R2, a cell surface receptor that is selectively expressed in glioma cells, but not normal brain cells or other tissues. We demonstrated that engineered single-chain antibody retains an exclusive specificity as well as a high affinity to IL13R2, and successfully re-targets engineered adenovirus and therapeutic CAR T cells to IL13R2- expressing glioma cells in pre-clinical models of GBM, in vitro and in vivo. In addition to being overexpressed in the majority of GBMs, IL13R2 expression has been associated with the highly aggressive mesenchymal subtype gene expression signature and poorer patient prognosis, all of which suggest that targeting IL13R2- expressing glioma cells could improve GBM patient outcomes. We hypothesize that NSCs engineered to secrete bi-specific tandem IL13Rα2xCD3 scFv antibody (biscFvNSCs) will promote anti-tumor immune response through the activation and engagement of T cells with GBM cells. Advancing this therapeutic for clinical application will be accomplished through R21 phase, during which we will focus on the detailed analysis and characterization of biscFvNSCs for production of functional biscFv IL13Rα2xCD3 and the ability to activate T cells and elicit cytotoxic effect against IL13Rα2-expressing glioma cells in vitro. During R33 phase, we will evaluate functional responses of biscFvNSCs in vivo, using immune-competent and patient-derived xenograft models of GBM. This will include the ability of biscFvNSCs to locally produce biscFv IL13Rα2xCD3, engage T and glioma cells, and elicit potent anti-glioma activity. During each phase, we will achieve quantitative milestones, which will further ensure the optimization of biscFvNSCs as a new therapeutic modality for GBM treatment.

胶质母细胞瘤（GBM）是成人中最具侵袭性的恶性脑癌。被诊断患有以下疾病的人 GBM 的治疗选择有限且预期生存期短。现有的一个主要问题 治疗方法的缺点是缺乏针对肿瘤细胞的特异性，这导致大量的治疗 毒性。抗体介导的肿瘤相关抗原的特异性靶向已成为治疗癌症的成功策略 癌症治疗，因为它限制了全身输注药物的脱靶效应。此类基因改造 抗体与有效的递送策略相结合可以大大提高这些抗体的抗肿瘤功效 分子。其中一种修饰是双特异性串联单链抗体 (biscFv)，可促进 T 细胞 肿瘤细胞相互作用，进而杀死肿瘤细胞。然而，biscFv 半衰期短且速度快 清除，需要频繁或连续输注才能达到治疗效果。我们建议 通过产生产生 biscFv 的神经干细胞 (NSC) 来克服这些障碍。 NSC 有能力 在全身、局部和鼻内递送后追踪脑肿瘤细胞，并有效地递送治疗 GBM 临床前模型中肿瘤部位的有效负载。分泌biscFv的NSC可以直接与 患者自体 T 细胞产生局部免疫反应，旨在根除肿瘤。 最近，我们开发并表征了一种专门针对细胞表面 IL13Rα2 的单克隆抗体。 选择性在神经胶质瘤细胞中表达的受体，但在正常脑细胞或其他组织中不表达。我们 证明工程单链抗体保留了独特的特异性以及对 IL13Rα2，并成功将工程腺病毒和治疗性 CAR T 细胞重新靶向 IL13Rα2- 在 GBM 临床前模型中体外和体内表达神经胶质瘤细胞。除了过度表达之外 在大多数 GBM 中，IL13Rα2 表达与高度侵袭性间充质细胞相关。 亚型基因表达特征和较差的患者预后，所有这些都表明靶向 IL13Rα2- 表达神经胶质瘤细胞可以改善 GBM 患者的预后。我们假设 NSC 被设计为 分泌双特异性串联IL13Rα2xCD3 scFv抗体（biscFvNSCs）将促进抗肿瘤免疫 通过 T 细胞的激活和与 GBM 细胞的接触来做出反应。推进这一治疗 临床应用将通过R21阶段完成，在此期间我们将重点关注详细的 用于生产功能性 biscFv IL13Rα2xCD3 的 biscFvNSC 的分析和表征以及 在体外激活 T 细胞并对表达 IL13Rα2 的神经胶质瘤细胞产生细胞毒性作用。在R33阶段， 我们将使用免疫活性和患者来源的细胞来评估 biscFvNSC 的体内功能反应 GBM 异种移植模型。这将包括 biscFvNSC 本地产生 biscFv IL13Rα2xCD3 的能力， 参与 T 细胞和神经胶质瘤细胞，并引发有效的抗神经胶质瘤活性。在每个阶段，我们将实现 定量里程碑，这将进一步确保 biscFvNSC 作为新治疗方式的优化 用于 GBM 治疗。