Immune cell control of ovarian cancer

目标1：单核细胞已经在化学诱导的小鼠癌症模型中表明，干扰素(IFN)在肿瘤的免疫监视和免疫编辑中起着重要作用。虽然干扰素α(IFNA)和伽马(IFNG)在体外被证明具有强大的抗肿瘤和抗增殖特性，但它们在临床上表现出的效果很小。我们的观察表明，在单核细胞存在的情况下，IFN在杀伤肿瘤细胞方面发挥着重要作用，这表明单核细胞和IFN联合治疗可能比单独使用IFN具有更有效的效果。我们和其他人已经证明，当与从外周血中分离的单核细胞结合使用时，干扰素是更有效的抗癌药物。在这些研究的基础上，我们完成了免疫细胞治疗的第一阶段临床试验，使用IFN刺激的体外单核细胞自体转移到具有抵抗力疾病的患者的腹膜腔内。我们现在正在确定干扰素诱导的单核细胞杀伤卵巢癌细胞的机制。我们发现，IFN诱导了一组在单核细胞中受调控的独特基因，这种基因只有在IFNA和IFNG结合时才会发生，而且这个信号可能是由转录因子MZF1驱动的。为了更好地模拟卵巢癌的腹膜微环境，我们从细胞培养株和原代标本中建立了一种新的生长复杂的卵巢癌肿瘤的模型。我们能够成像和量化IFN刺激的单核细胞和肿瘤生长之间的复杂相互作用，为研究功能蛋白质组相互作用提供了一个优越的模型。此外，在我们的第一阶段试验中，我能够检查接受IFN刺激的腹膜内自体单核细胞的妇女的组织和腹腔液样本，为在完全免疫反应的背景下确定双干扰素诱导的单核细胞活动机制提供了另一个平台。目的2：研制抗人肿瘤组织膜蛋白半纯化制剂ADCC A的单抗。这种蛋白质制剂在之前的临床试验中被用作癌症疫苗，在那里它被证明是安全有效的。抗体NEO201被证明与免疫抗原制剂以及几个人类肿瘤细胞系和来自结直肠癌、胰腺癌、肺癌和卵巢癌患者的组织发生反应。NEO201与正常人体组织没有明显的交叉反应，因此是一种潜在的治疗产品。研究表明，NEO201的靶点与CEACAM-5/6有关，CEACAM-5/6是癌胚抗原蛋白家族的成员，已被证明与几种癌症类型有关。子宫内膜癌、乳腺癌和卵巢癌在人类肿瘤样本中的表达明显增加。在子宫内膜癌中，45/88(51%)的组织样本通过免疫组织化学显示为阳性，38/72(53%)的乳腺组织中有38/72(53%)呈阳性反应，尽管在本系列中有16/129(12%)的卵巢癌标本呈阳性，但在代表600多个样本的卵巢癌组织阵列的IHC中，粘液性15/22(68%)和印戒细胞2/2(100%)两种亚型显示显著的反应性(50%)。我们的第一阶段临床试验正在进行中，以展示NEO201在可能表达靶点的实体肿瘤患者中的安全性和初步活性。目的3：T细胞间皮蛋白(MESO)是一种41kD的细胞表面糖蛋白，在多种人类肿瘤中高表达，包括卵巢高级别浆液性腺癌(75%)、胰腺癌(85%)、三阴性乳腺癌(66%)、上皮样间皮瘤(95%)等。虽然Meso在正常细胞上的功能不是必需的，但肿瘤细胞上Meso的表达可能参与了肿瘤的病理过程，高表达与较差的预后、增加转移扩散和激活细胞生长途径有关。一种非常创新的免疫治疗方法是使用嵌合抗原受体修饰的T细胞(CAR)。CAR T细胞疗法依赖于重新设计自体T细胞来表达一种受体，使T细胞能够识别肿瘤细胞。CAR是由细胞外抗原结合域和细胞内T细胞信号域组成的重组受体。当在T细胞中表达时，CARS将T细胞重定向至以非人类白细胞抗原(HL A)方式表达靶抗原的癌细胞。最广泛使用的T细胞修饰方法是病毒转导、整合和表达表达嵌合受体的基因结构。产生CAR T细胞疗法的另一种方法可能提供强大的抗肿瘤活性，并改善安全性和产品制备，涉及使用信使核糖核酸修饰T细胞。使用信使核糖核酸重新设计患者的T细胞以表达肿瘤抗原靶向的CAR T细胞可以在几个小时内完成，从而允许现场准备和部署到多个治疗地点。MRNACAR T细胞的安全因素是寿命有限，半衰期与抗体疗法相似，而且缺乏快速的免疫激活和增殖，限制了严重细胞因子释放副作用的风险。通过肿瘤内、腹膜内和静脉给药途径，使用信使核糖核酸的中位靶向CAR T细胞在临床前研究和临床研究中显示出重要的前景。我们正在进行一项1期临床试验，测试CARMA MCY-M11在卵巢癌和腹膜癌患者中腹膜内给药的安全性。

Goal 1: Monocytes It has been shown in mouse models of chemically induced cancer that Interferons (IFNs) are important in the immune-surveillance and immune-editing of tumors. While IFNs alpha (IFNa) and gamma (IFNg) have been shown to have potent anti-neoplastic and anti-proliferative properties in vitro, they have shown little efficacy in the clinic. Our observations that IFNs play an important role in killing tumor cells in the presence of monocytes suggests that combination therapy of monocytes and IFNs may have a more potent effect than IFNs alone. We and others have shown that IFNs are more potent anti-cancer agents when used in combination with monocytes isolated from the peripheral blood. Based on these studies we have completed a Phase 1 clinical trial of immune cell therapy using autologous transfer of ex vivo monocytes stimulated with IFNs into the peritoneal cavity of patients who have resistant disease. We are now in the process of defining the mechanism of IFN-induced monocyte killing of ovarian cancer cells. we found that IFNs induce a unique set of genes regulated in monocytes that occurs only with the combination of IFNa and IFNg, and this signature may be driven by the transcription factor MZF1. In order to better mimic the peritoneal microenvironment of ovarian cancer, we developed a new model of growing complex ovarian cancer neoplasms from cell culture lines and primary samples. We are able to image and quantify complex interactions of IFNs-stimulated monocytes and cancer growths, providing a superior model for studying functional proteomic interactions. In addition, I have the ability to examine tissue and peritoneal fluid samples from women on our phase 1 trial receiving intraperitoneal autologous monocytes stimulated with IFNs, providing another platform for defining dual-IFN induced mechanisms of monocyte activity in the context of the complete immune response. Goal 2: ADCC A monoclonal antibody was developed against a semi-purified human membrane protein preparation derived from cancer tissues. The protein preparation was used in previous clinical trials for use as a cancer vaccine, where it was demonstrated to be safe and efficacious. The antibody, NEO201, was shown to react with the immunizing antigen preparation, as well as several human tumor cell lines and tissues from colorectal, pancreas, lung, and ovarian cancer patients. NEO201did not cross-react significantly with normal human tissues, thus representing a potential therapeutic product. The target of NEO201was studied and shown to be related to CEACAM-5/6, a member of the carcinoembryonic antigen family of proteins, which has been shown to be associated with several cancer types. Endometrial, breast and ovarian cancer have specifically been found to have increased expression in human tumor samples. In endometrial cancer, 45/88 (51%) of tissue samples show reactivity through immunohistochemistry, 38/72 (53%) of breast, and although 16/129 (12%) of ovarian cancer specimens stain positive in this series, two subtypes, mucinous 15/22 (68%)and signet cell 2/2 (100%) ovarian cancers, shows significant reactivity (50%) in an IHC of ovarian cancer tissue arrays representing over 600 samples. Our phase 1 clinical trial is ongoing to demostrate safety and preliminary activity of the NEO201 in patients with solid tumors likely to express the target. Goal 3: T cells Mesothelin (MESO) is a 41-kD cell surface glycoprotein that is highly expressed in many human cancers, including high grade serous adenocarcinoma of the ovary (75%), pancreatic adenocarcinoma (85%), triple negative breast cancer (66%), epitheliod mesothelioma (95%) of patients with MESO-expressing malignancies. While the function of MESO on normal cells is non-essential, the expression of MESO on cancer cells may contribute to the pathology of cancer, with higher expression associated with poorer prognosis, increased metastatic spread, and activation of cell growth pathways. A tremendously innovative immunotherapeutic approach is the use of chimeric antigen receptor-modified T cells (CAR). CAR T-cell therapy relies on re-engineering autologous T cells to express a receptor that allows the T cells to recognize tumor cells. A CAR is a recombinant receptor composed of an extracellular antigen-binding domain and an intracellular T-cell signaling domain. When expressed in T cells, CARs redirect the T cells to target the cancer cells that express the targeted antigen in a human leukocyte antigen (HLA)-independent manner. The most widely used method for T-cell modification is viral transduction, integration and expression of a genetic construct that expresses the chimeric receptor. Another approach to the generation of CAR T-cell therapies that may provide potent anti-tumor activity and improve safety and product preparation involves the use of mRNA to modify T-cells. Using mRNA to re-engineer a patient's T-cells to express a tumor-antigen targeted CAR T-cell can be accomplished in a few hours, allowing on-site preparation and deployment to multiple treatment locations. mRNA CAR T-cells have the safety factor of a limited lifespan, with half-life times similar to antibody therapeutics, and lack of rapid immune activation and proliferation, limiting the risk for severe cytokine release side effects. Meso-targeted CAR T-cells using mRNA have demonstrated significant promise in preclinical studies and clinical studies by intratumoral, intraperitoneal and intravenous of routes of administration. We are running a phase 1 clinical trial testing the safety of intraperitoneal administration of the CARMA MCY-M11 in women with ovarian cancer and peritoneal carcinomatosis.