Immunotherapy for Malignant Mesothelioma and Lung Cancer

恶性间皮瘤和肺癌的免疫治疗

• 批准号: 10702415
• 负责人: RAFFIT HASSAN
• 金额: $ 201.88万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10702415
• 关键词: Adoptive Cell Transfers; Affinity; Animal Model; Antibodies; Ascites; Bacterial Toxins; Biopsy; C-terminal; CAR T cell therapy; CTLA4 gene; Cancer Patient; Categories; Cell Line; Cell surface; Cells; Characteristics; Clinic; Clinical; Clinical Trials; Complex; Conduct Clinical Trials; DNA Repair Gene; Development; Differentiation Antigens; Distal; Drug Targeting; Enrollment; Epidermal Growth Factor Receptor; Epitopes; Follow-Up Studies; Genetic; Germ-Line Mutation; Goals; Human; Immune; Immune checkpoint inhibitor; Immunocompetent; Immunotherapeutic agent; Immunotherapy; Immunotoxins; In Vitro; Infiltration; Inflammation; Injections; KRAS2 gene; Laboratories; Laboratory Research; Laboratory Study; Lead; Link; Liquid substance; Longterm Follow-up; Lung Adenocarcinoma; Malignant Neoplasms; Malignant mesothelioma; Malignant neoplasm of lung; Malignant neoplasm of ovary; Malignant neoplasm of thorax; Maximum Tolerated Dose; Membrane; Mesothelial Cell; Mesothelioma; Modeling; Monoclonal Antibodies; Morphology; Mus; Mutation; Non-Small-Cell Lung Carcinoma; Oryctolagus cuniculus; Outcome; Pancreatic Adenocarcinoma; Pathway interactions; Patient Recruitments; Patients; Peripheral Blood Mononuclear Cell; Peritoneum; Pharmaceutical Preparations; Phase I Clinical Trials; Phase I/II Clinical Trial; Platinum; Pleura; Pleural; Pleural Mesothelioma; Prior Therapy; Prognosis; Pseudomonas aeruginosa toxA protein; Publishing; Regulation; Research; Role; Safety; Screening for cancer; Surface; T cell therapy; T-Cell Activation; T-Cell Receptor; Therapeutic Agents; Translational Research; Tumor Cell Line; Tumor-Derived; Vaccines; Work; anti-PD-L1 antibodies; anti-PD1 antibodies; antitumor effect; cancer prevention; cancer therapy; cell killing; chimeric antigen receptor; chimeric antigen receptor T cells; clinical development; cytotoxicity test; de-immunization; differential expression; drug sensitivity; effective therapy; falls; graft vs host disease; human model; humanized mouse; immune checkpoint; immunotherapy clinical trials; improved; in vivo; in vivo Model; ipilimumab; mesothelin; mouse model; mutant; neoplastic cell; novel; novel strategies; novel therapeutics; pembrolizumab; pericardial sac; phase 2 study; phase I trial; phase II trial; preclinical study; programmed cell death ligand 1; programs; systemic inflammatory response; targeted agent; translational medicine; tumor; tumor xenograft; tumor-immune system interactions; vector

1. Exploiting mesothelin for mesothelioma therapy and related translational research Our current studies are focused on using immunotherapy directed against the tumor differentiation antigen mesothelin, which is expressed on normal mesothelial cells lining the pleura, pericardium, and peritoneum, but is highly expressed in several human tumors especially mesothelioma, ovarian cancer, lung cancer and pancreatic adenocarcinomas. This differential expression of mesothelin makes it an attractive candidate for tumor specific therapy. Our efforts are now focused on exploiting it for mesothelioma therapy using different approaches. These include anti mesothelin immunotoxin (LMB-100), an anti-mesothelin drug conjugate (BAY 94-9343), mesothelin vaccine (CRS-207) and adoptive T-cell therapy (TC-210). We are currently developing mesothelin-targeting adoptive cellular therapy, using chimeric antigen receptor (CAR)-T cells. Majority of the anti-mesothelin antibodies in clinical development target the membrane distal region of mesothelin, that could partly account for the lack of activity of anti-mesothelin CAR-T cell therapy in the clinic. To improve CAR-T cell anti-tumor activity, we propose a new approach i.e. developing anti-mesothelin CAR-T cells that target an epitope close to the surface of tumor cells. My collaborator, Dr. Mitchell Ho, has identified a high affinity rabbit monoclonal antibody (YP218) specific for region III, which is located at the C-terminal end of mesothelin close to the tumor cell surface. We have tested the cytotoxicity of the hYP218 CAR-T construct on several mesothelin expressing cell lines and anti-tumor effect in animal models. The pre-clinical studies have shown increased tumor killing (1). This construct is now under clinical development. Parallelly, we are conducting a clinical trial of a T-cell receptor fusion construct (TRuCs). Unlike other constructs, TRuCs are naturally incorporated into the native TCR complex, thus exploiting the full potential of TCR-driven T cell activation, effector function, and regulation. LMB-100 is an immunotoxin consisting of the anti-mesothelin Fv linked to a truncated form of the potent bacterial toxin, Pseudomonas exotoxin A, which has been de-immunized to decrease its antigenicity. We have recently completed the phase I trial of LMB-100 and established its safety and maximum tolerated dose (MTD), published in Cancer (2). Currently, we are evaluating the results of a recently concluded a phase II study of LMB-100 in patients with mesothelioma, in combination with immune checkpoint inhibitor Pembrolizumab. Since LMB-100 causes systemic inflammation and increase in immune cell infiltration in patient tumors, we hypothesized that intra- tumoral injection of LMB-100 would lead to increased inflammation and immune cell infiltration. Administering checkpoint inhibitors would further increase tumor-cell killing. We are presently conducting a phase 1 clinical trial in patients with mesothelioma where they are intratumorally administered LMB-100 on days 1 and 4 followed by CTLA-4 checkpoint inhibitor, ipilimumab, given i.v. on day 2 of a 21 day cycle. Patients will receive 2 cycles of LMB-100 plus ipilimumab, followed by 2 cycles of ipilimumab alone. Tumor biopsies will be performed prior to each administration of LMB-100, to evaluate changes in the tumor immune microenvironment. The trial is recruiting patients. We have previously shown that germline mutations in DNA repair genes increases sensitivity to platinum therapy and improves overall survival in patients with pleural mesothelioma (3). Currently, my laboratory is studying germline mutations in DNA repair genes that could predispose to mesothelioma and influence clinical outcome. We are enrolling patients and their relatives harboring such mutations for a long term follow up study, for early cancer detection and prevention. In the laboratory, we have focused on developing in-vitro and in-vivo models of human mesothelioma. We have established several early passage tumor cell lines from ascites and pleural fluid of patients. We have evaluated the morphological and genetic characteristics of these cell lines and are using them to study in-vitro drug sensitivity. Additionally, we have established a humanized mesothelioma xenograft tumor model with patient derived tumor cells and human PBMCs from healthy donor for in-vivo studies. As the development of Graft Versus Host Disease (GVHD) in the PBMC-humanized mouse model limits assessment of duration of anti-tumor efficacy, we have developed a syngeneic immunocompetent mouse model. Because the immunotoxin LMB-100 can target human mesothelin specifically, we established a human mesothelin expressing immunocompetent syngeneic mouse tumor model by transfecting PD-L1 positive mouse lung adenocarcinoma cell line with a hMSLN expressing vector encoding the membrane bound fragment of hMSLN. These cell lines were used to develop tumor. We have studied the effect of LMB-100 in combination with anti-PD1 antibody in both the models and have seen tumor regression. We have published our findings in Science Translational Medicine (4). These models are essential to evaluate novel therapeutic agents for mesothelioma and for the mechanistic studies of anti-tumor efficacy. Other ongoing laboratory studies are focused on understanding the mesothelioma tumor immune micro-environment and changes following treatment with anti-mesothelin targeted agents. 2. Immunotherapy to treat lung cancers. We are currently conducting clinical trial of the anti-PD-L1 monoclonal antibody MSB0010718C in patients with lung adenocarcinoma who have failed prior therapies. Our laboratory has recently shown that about 25% of patients with metastatic lung adenocarcinoma highly express mesothelin. Mesothelin expression in these tumors is highly associated with KRAS mutations and wild type EGFR status and is, independently, associated with poor prognosis. Our hypothesis is that patients with K-RAS mutant lung cancer can benefit from mesothelin directed therapies. Clinical trials of mesothelin directed therapies for treating lung cancer are about to open. Our laboratory is also studying the role of immune checkpoints in malignant mesothelioma so that drugs targeting this pathway can be exploited for treating mesothelioma. We are currently conducting a Phase II trial of NSCLC patients treated with LMB-100 in combination with pembrolizumab.

1. 利用间皮素进行间皮瘤治疗和相关转化研究我们目前的研究重点是使用针对肿瘤分化抗原间皮素的免疫疗法，间皮素在胸膜、心包和腹膜内衬的正常间皮细胞上表达，但在几种人类肿瘤中高表达，特别是间皮瘤、卵巢癌、肺癌和胰腺癌 腺癌。间皮素的这种差异表达使其成为肿瘤特异性治疗的有吸引力的候选者。我们现在的工作重点是使用不同的方法将其用于间皮瘤治疗。这些包括抗间皮素免疫毒素 (LMB-100)、抗间皮素药物缀合物 (BAY 94-9343)、间皮素疫苗 (CRS-207) 和过继性 T 细胞疗法 (TC-210)。我们目前正在开发使用嵌合抗原受体 (CAR)-T 细胞的间皮素靶向过继细胞疗法。临床开发中的大多数抗间皮素抗体主要针对间皮素的膜远端区域，这可能部分解释了抗间皮素 CAR-T 细胞疗法在临床上缺乏活性的原因。为了提高 CAR-T 细胞的抗肿瘤活性，我们提出了一种新方法，即开发针对靠近肿瘤细胞表面的表位的抗间皮素 CAR-T 细胞。我的合作者 Mitchell Ho 博士鉴定出了一种针对 III 区的高亲和力兔单克隆抗体 (YP218)，该区位于靠近肿瘤细胞表面的间皮素 C 末端。我们测试了 hYP218 CAR-T 构建体对几种间皮素表达细胞系的细胞毒性以及动物模型中的抗肿瘤作用。临床前研究表明肿瘤杀伤作用增强 (1)。该结构目前正处于临床开发阶段。与此同时，我们正在进行 T 细胞受体融合构建体 (TRuC) 的临床试验。与其他构建体不同，TRuC 自然地融入天然 TCR 复合物中，从而充分发挥 TCR 驱动的 T 细胞激活、效应功能和调节的潜力。 LMB-100 是一种免疫毒素，由抗间皮素 Fv 与截短形式的强效细菌毒素假单胞菌外毒素 A 组成，该毒素已被去免疫以降低其抗原性。我们最近完成了 LMB-100 的 I 期试验，并确定了其安全性和最大耐受剂量 (MTD)，发表在《Cancer》(2) 上。目前，我们正在评估最近结束的一项 LMB-100 与免疫检查点抑制剂 Pembrolizumab 联合治疗间皮瘤患者的 II 期研究的结果。由于LMB-100会引起全身炎症并增加患者肿瘤中的免疫细胞浸润，我们假设肿瘤内注射LMB-100会导致炎症和免疫细胞浸润增加。施用检查点抑制剂将进一步增加肿瘤细胞的杀伤力。 我们目前正在对间皮瘤患者进行一项 1 期临床试验，在第 1 天和第 4 天瘤内注射 LMB-100，然后静脉注射 CTLA-4 检查点抑制剂伊匹单抗 (ipilimumab)。在 21 天周期的第 2 天。患者将接受 2 个周期的 LMB-100 加伊匹单抗治疗，然后接受 2 个周期的单独伊匹单抗治疗。每次施用 LMB-100 之前都会进行肿瘤活检，以评估肿瘤免疫微环境的变化。该试验正在招募患者。我们之前已经表明，DNA 修复基因的种系突变会增加胸膜间皮瘤患者对铂类治疗的敏感性并提高总体生存率 (3)。目前，我的实验室正在研究 DNA 修复基因的种系突变，这些突变可能诱发间皮瘤并影响临床结果。我们正在招募携带此类突变的患者及其亲属进行长期随访研究，以进行早期癌症检测和预防。在实验室，我们专注于开发人类间皮瘤的体外和体内模型。我们已经从患者的腹水和胸水中建立了几种早期传代肿瘤细胞系。 我们评估了这些细胞系的形态和遗传特征，并利用它们来研究体外药物敏感性。此外，我们还利用患者来源的肿瘤细胞和来自健康供体的人 PBMC 建立了人源化间皮瘤异种移植肿瘤模型，用于体内研究。由于PBMC人源化小鼠模型中移植物抗宿主病（GVHD）的发展限制了抗肿瘤功效持续时间的评估，我们开发了同基因免疫活性小鼠模型。由于免疫毒素LMB-100可以特异性靶向人间皮素，我们通过用编码hMSLN膜结合片段的hMSLN表达载体转染PD-L1阳性小鼠肺腺癌细胞系，建立了表达人间皮素的免疫活性同系小鼠肿瘤模型。这些细胞系用于形成肿瘤。我们在两个模型中研究了 LMB-100 与抗 PD1 抗体联合的效果，并看到肿瘤消退。我们已在《科学转化医学》(4) 上发表了我们的研究结果。这些模型对于评估新型间皮瘤治疗药物和抗肿瘤功效的机制研究至关重要。其他正在进行的实验室研究重点是了解间皮瘤肿瘤免疫微环境以及抗间皮素靶向药物治疗后的变化。 2. 治疗肺癌的免疫疗法。我们目前正在对既往治疗失败的肺腺癌患者进行抗PD-L1单克隆抗体MSB0010718C的临床试验。我们实验室最近显示，约25%的转移性肺腺癌患者高表达间皮素。这些肿瘤中的间皮素表达与 KRAS 突变和野生型 EGFR 状态高度相关，并且独立地与不良预后相关。我们的假设是，K-RAS 突变肺癌患者可以从间皮素定向治疗中受益。间皮素定向疗法治疗肺癌的临床试验即将启动。我们的实验室也在研究免疫检查点在恶性间皮瘤中的作用，以便开发针对该途径的药物来治疗间皮瘤。我们目前正在进行一项针对 NSCLC 患者的 II 期试验，该患者接受 LMB-100 联合派姆单抗治疗。