Enhancing cell therapy for brain tumors

摘要 多形性胶质母细胞瘤（GBM）是最致命的原发性脑癌，标准治疗基于 手术、放疗和化疗可促进约15个月的总生存期。这 导致人们对基于免疫的方法重新产生兴趣。虽然增强免疫力是成功的 在其他恶性肿瘤如黑色素瘤和白血病/淋巴瘤中，它们在治疗脑肿瘤中的功效 非常有限。肿瘤抗原异质性，肿瘤部位浸润淋巴细胞数量有限， 以及检查点抑制剂药物不能穿过血脑屏障都是 GBM的有效免疫疗法。GBM患者的免疫力改变进一步导致穷人 这些肿瘤及其对疫苗和检查点抑制剂的相对耐药性。利用健康 以过继细胞疗法形式的供体免疫细胞可能提供更有效的替代方案。但受限 可以获得关于效应物群体的特性的信息，所述效应物群体在肿瘤中发挥有效的抗肿瘤活性。 大脑约翰霍普金斯大学的里克·琼斯博士利用单倍相合供体 脑肿瘤的单倍骨髓移植提供了一个机会，以测试输注的健康供体免疫的动力学 细胞，提供了供体免疫细胞可以对抗脑肿瘤的一瞥。 以前的经验与单倍骨髓移植表明，效应细胞的先天免疫-自然杀伤细胞 (NKs)和不变的自然杀伤T细胞（iNKT）介导的抗肿瘤活性， 复发先天免疫细胞在脑肿瘤环境中也是潜在有利的，这提供了非常好的治疗效果。 对与（例如）基于CD 19-CAR T细胞的疗法相关的不良事件几乎没有耐受性。最后， iNKT和NK细胞都可以在趋化因子CCL 2梯度后主动迁移到GBM部位， 由肿瘤细胞和周围的肿瘤相关巨噬细胞（TAM）释放。萨沃尔多博士在 北卡罗来纳州大学在嵌合抗原受体（CAR）的开发方面拥有独特的专业知识 转导的T细胞和iNKT细胞和博士Bollard在儿童国家有丰富的经验， 基因修饰的T细胞和NK细胞。因此，在这个合作提案中，我们假设， 来自健康供体的非耐受化先天免疫细胞（NK和iNKT）将促进抗- GBM患者的肿瘤免疫力，并可开发为有效的细胞疗法。这一总体 假设将在3个特定目标中进行检验，其中Jones博士（目标1）将比较 iNKT和从健康供体与GBM患者分离的NK细胞。在Aim 2中，Savoldo博士和 研究小组将评估健康供体iNKT和NK细胞的抗肿瘤功效，这些细胞被修饰以表达抗肿瘤抗体。 B7 H3嵌合抗原受体（CAR），而在Aim 3中，CNMC的Bollard和Cruz博士克服了抑制性B7 H3嵌合抗原受体（CAR）。 通过靶向免疫抑制性肿瘤相关巨噬细胞（TAM）， 转化生长因子β

ABSTRACT Glioblastoma multiforme (GBM) is the most lethal primary brain cancer, with standard treatments based on surgery, radiotherapy, and chemotherapy promoting an overall survival of approximately 15 months. This has led to a resurgence of interest in immune-based approaches. While augmenting immunity has been successful in other malignancies like melanoma and leukemia/lymphoma, their efficacy in treatment of brain tumors has been very limited. Tumor antigen heterogeneity, limited number of infiltrating lymphocytes at the tumor site, and failure of checkpoint inhibitor drugs to cross the blood brain barrier all represent major obstacles to effective immune-therapies in GBM. Altered immunity in patients with GBM further contributes to the poor prognoses of these tumors and their relative resistance to vaccine and checkpoint inhibitors. Utilizing healthy donor immune cells in the form of adoptive cell therapy may offer a more effective alternative. However, limited information is available on the properties of effector populations that would exert effective anti-tumor activity in the brain. Pioneering work by Dr. Rick Jones at Johns Hopkins University utilizing haploidentical donor transplant (haplo-BMT) for brain tumors offers an opportunity to test kinetics of infused healthy donor immune cells in this patient population, providing a glimpse of what donor immune cells can do against brain tumors. Previous experiences with haplo-BMT suggest that effector cells of the innate immunity – natural killer cells (NKs) and invariant natural killer T cells (iNKTs) – mediate anti-tumor activity with decreased frequency of relapse. Innate immune cells are also potentially advantageous in the brain tumor setting, which affords very little tolerance for the adverse events associated with (for example) CD19-CAR T cell based-therapies. Finally, both iNKT and NK cells can actively migrate to the site of GBM following the CCL2 gradient, the chemokine released by tumor cells and the surrounding tumor associated macrophages (TAM). Dr. Savoldo at University of North Carolina has unique expertise in the development of chimeric antigen receptor (CAR) transduced T cells and iNKT cells and Dr. Bollard at Children's National has extensive experience with the genetically modified T cells and NK cells. Hence, in this collaborative proposal we hypothesize that nontolerized innate immune cells (NKs and iNKTs) derived from healthy donors will promote anti- tumor immunity in patients GBM, and may be developed as effective cellular therapies. This overarching hypothesis will be tested in 3 Specific Aims where Dr. Jones (Aim 1) will compare the cellular signatures of iNKTs and NK cells isolated from healthy donors versus patients with GBM. In Aim 2, Dr. Savoldo and the UNC team will evaluate the anti-tumor efficacy of healthy donor iNKT and NK cells modified to express an anti- B7H3 chimeric antigen receptor (CAR) and in Aim 3, Drs. Bollard and Cruz at CNMC overcome the inhibitory tumor microenvironment in GBM by targeting immune suppressive tumor associated macrophages (TAMs) and TGFβ.