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，标准的治疗方法是基于 手术、放射治疗和化疗可使患者的总体生存时间延长约15个月。这有 导致了人们对基于免疫的方法的兴趣死灰复燃。虽然增强免疫力取得了成功 在其他恶性肿瘤中，如黑色素瘤和白血病/淋巴瘤，它们在治疗脑肿瘤方面的疗效 一直非常有限。肿瘤抗原异质性，肿瘤部位浸润性淋巴细胞数量有限， 检查点抑制剂药物未能通过血脑屏障都是主要障碍 基底膜的有效免疫治疗。GBM患者免疫功能的改变进一步加剧了贫困 这些肿瘤的预后及其对疫苗和检查点抑制剂的相对耐药性。利用健康 过继细胞疗法形式的供者免疫细胞可能提供一种更有效的替代方案。然而，有限的 关于将在体内发挥有效的抗肿瘤活性的效应者群体的特性的信息可用 大脑。约翰·霍普金斯大学里克·琼斯博士利用半相合捐赠者所做的开创性工作 脑肿瘤移植(haplo-bmt)提供了一个测试输注健康供者免疫的动力学的机会。 这一患者群体中的细胞，使人们得以一窥捐赠者免疫细胞对脑瘤的作用。 以往的半骨髓移植经验表明，先天免疫的效应细胞--自然杀伤细胞 (NKS)和不变自然杀伤T细胞(INKT)-介导抗肿瘤活性，频率降低 旧病复发。先天免疫细胞在脑瘤环境中也有潜在的优势，这提供了非常 对(例如)CD19-CAR T细胞疗法相关的不良事件几乎没有耐受性。最后， 趋化因子CCL2梯度后，iNKT和NK细胞都能主动迁移到GBM部位 由肿瘤细胞和周围的肿瘤相关巨噬细胞释放()。萨沃尔多博士在 北卡罗来纳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，儿童国家医院的Bollard博士在 转基因T细胞和NK细胞。因此，在这份合作提案中，我们假设 来自健康捐赠者的非耐受性先天免疫细胞(NKs和iNKT)将促进抗- 肿瘤免疫在患者的GBM，并可能被开发为有效的细胞治疗。这是最重要的 假设将在三个特定的目标中进行测试，琼斯博士(目标1)将比较 从健康献血者和GBM患者分离的iNKT和NK细胞。在《目标2》中，萨沃尔多博士和 UNC团队将评估健康捐赠者iNKT和表达抗肿瘤基因的NK细胞的抗肿瘤效果 在B7H3嵌合抗原受体(CAR)和AIM 3中，CNMC的Bollard和Cruz博士克服了这种抑制 靶向免疫抑制肿瘤相关巨噬细胞(TAMs)和TAMs的GBM肿瘤微环境 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.