ADMINISTRATION OF HER2 CHIMERIC RECEPTOR AND TGFBETA DOMINANT NEGATIVE RECEPTOR

HER2嵌合受体和TGFβ显性阴性受体的施用

• 批准号: 8356777
• 负责人: Stephen Gottschalk
• 金额: $ 1.07万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8356777
• 关键词: Animal Model; Antigen Receptors; Antigens; Autologous; Autologous Epstein-Barr Virus-Specific Cytotoxic T Lymphocytes; Biological Response Modifiers; Blood; Blood specimen; Cancer Patient; Cell physiology; Cells; Clinic; Clinical Research; Clinical Trials; Cytotoxic T-Lymphocytes; Disease; Dominant Negative Receptor; Dominant-Negative Mutation; ERBB2 gene; Engineering; Environment; FDA approved; Funding; Gene Transfer; Gene-Modified; Genes; Grant; Growth Factor Receptors; Hour; Human Herpesvirus 4; Immune; Immunity; Immunosuppression; Immunotherapy; Infusion procedures; Liquid substance; Lymphoma; Malignant Neoplasms; Malignant neoplasm of lung; Measures; Metastatic/Recurrent; Modification; Moloney Leukemia Virus; Monitor; National Center for Research Resources; Outcome; Patients; Population; Principal Investigator; Research; Research Infrastructure; Resistance; Resources; Retroviral Vector; Safety; Secondary to; Source; Stream; Structure; Subgroup; Synthetic Genes; T-Lymphocyte; Technology; Tissue Sample; Transforming Growth Factor beta; Transforming Growth Factors; Tumor Tissue; United States National Institutes of Health; Veins; Virus; cancer cell; cancer therapy; cost; experience; fighting; human TGFBR2 protein; immune function; intravenous injection; killer T cell; killings; lymphocyte proliferation; meetings; neoplastic cell; peripheral blood; pre-clinical; preclinical study; receptor; receptor expression; safety testing; tumor; vector

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. ABSTRACT The outcome for patients with recurrent/metastatic lung cancer has not changed over the last 2 decades and most patients die of their disease. New anti-lung cancer therapies are needed and immunotherapy holds the promise to fulfill this need. The bodys immune defense against cancers often fails because cancers either do not induce or actively inhibit immunity. We will counteract these limitations by i) engineering killer T cells or recognize structures on lung cancer cells and ii) to resist the defenses imposed by the cancer cell environment. Using gene transfer technology, it may be possible to take advantage of the tumor killing ability of killer T cells and render them resistant to inhibitory factors secreted by tumor cells. We propose taking the patients own T cells and putting into them two genes. The first gene is for an artificial structure (receptor) that will direct the T cells to the tumor and the second gene is for a dominant negative receptor (DNR), which renders T cells resistant against one of the major inhibitory factors secreted by lung cancer cells. We will place the two genes into T cells that are pre-selected for their ability to recognize the Epstein Barr virus which is chronically present in most people. Because these EBV-specific T cells meet the virus positive cells as well as tumor cells, they receive extra stimulation. We will put the genes for the artificial receptor and the DNR into patients EBV-specific T cells using a modified virus (Moloney retroviral vector). These modified T-cells will then be given directly into the patients blood stream through a central line or a vein. Patients will be treated in the clinic and will be monitored closely for several hours after infusion. We will collect samples of blood from peripheral blood at regular intervals. We will look for the safety, the persistence and the function of the cells we put into the patients. Ultimately we hope to get evidence that these modified T cells are effective at fighting the cancer. I. HYPOTHESIS From our experience, the persistence of infused EBV-CTL in cancer patients is limited, most likely due to immune evasion strategies adapted by cancers. For example, lung cancers express high levels of TGF, a potent negative regulator of immune cells. In preclinical studies we have shown that EBV-CTL can be made resistant to the inhibitory effects of TGF by expressing a dominant negative TGF receptor II (DNR). DNR expressing CTL retained their effector function and had enhanced antitumor activity in preclinical animal models. Importantly, DNR expressing CTL did not show any evidence of autonomous (antigen-independent) CTL proliferation. These results provided the rationale for a RAC and FDA approved clinical trial (RAC #0507-724), in which we are testing the safety and efficacy of (TGF)-resistant CTL in lymphoma patients. In this study we will generate autologous EBV-specific CTL and genetically modify them using a retroviral vector expressing a dominant negative TGF beta receptor II (DNRII) to render them resistant to the immunosuppressive effects of TGF beta, and a chimeric antigen receptor recognizing HER2. Two separate vectors will be used to transfer the genes. Each cell product will thus contain: 1) Unmodified CTLs 2) HER2 CAR CTLs 3) TGF¿] DNR CTLs 4) HER2 CAR and TGF beta DNR CTLs We will adoptively transfer these CTLs to patients with advanced HER2-positive lung cancer. We will assess the safety of the infusion and measure the survival of the gene modified cells in subgroups 2, 3 and 4. We will detect each cell population by quantitative PCR and by immunostaining of peripheral blood and in tumor tissue/other fluids (when available). We anticipate that the CTL populations expressing TGF¿ DNR (subgroups 3 and 4) will survive better than the population lacking this modification (subgroup 2). We also anticipate survival of groups 3 and 4 will be equivalent, but if CAR expression itself has an adverse influence on CTL survival, then subgroup 3 will come to dominate. Alternatively, if cells expressing both TGF¿ DNR and the CAR localize better in tumors, then we would expect to see dominance of subgroup 4 in tissue samples. II. SPECIFIC AIMS Primary: To determine the safety of one intravenous injections of autologous Transforming Growth Factor (TGF)-resistant cytotoxic T lymphocytes (CTLs) directed to Epstein Barr virus (EBV) through their native receptor and HER2 through their chimeric antigen receptor (CAR) in patients with advanced HER2-positive lung cancers. Secondary: To compare the survival and the immune function of the TGF-resistant and non resistant components of the infused CAR-CTL. To assess the anti-tumor effects of the infused CAR-CTL.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 抽象的 在过去的20年中，复发/转移性肺癌的患者的结果没有改变，大多数患者死于疾病。需要新的抗肺癌疗法，免疫疗法有望满足这种需求。 对癌症的免疫反应通常会失败，因为癌症不会诱导或积极抑制免疫学。我们将通过i）工程杀手T细胞来抵消这些局限性，或识别肺癌细胞上的结构，ii）抵抗癌细胞环境施加的防御能力。使用基因转移技术，有可能利用杀伤剂T细胞的肿瘤杀伤能力，并使其对肿瘤细胞分泌的抑制因子具有抗性。我们提议将患者自己的T细胞带入两个基因。第一个基因是用于人工结构（受体），该结构将将T细胞引导到肿瘤，第二个基因用于显性阴性受体（DNR），该基因使T细胞具有抵抗由肺癌细胞分泌的主要抑制因子之一。 我们将将两个基因放入T细胞中，以识别大多数人长期存在的爱泼斯坦Barr病毒的能力。由于这些EBV特异性T细胞符合病毒阳性细胞以及肿瘤细胞，因此它们会受到额外的刺激。我们将使用修饰的病毒（Moloney逆转录病毒载体）将人工受体和DNR的基因和DNR的基因放入患者中。然后，这些修饰的T细胞将直接通过中央线或静脉直接将其血液流到患者中。患者将在诊所接受治疗，并在输注后几个小时对患者进行治疗。我们将定期从外周血中收集血液样本。我们将寻找我们投入患者细胞的安全性，持久性和功能。最终，我们希望获得证据表明这些修饰的T细胞有效地与癌症作斗争。 I.假设 根据我们的经验，癌症患者受感染的EBV-CTL的持久性受到限制，这很可能是由于癌症适应的免疫避难所策略所致。例如，肺癌表达高水平的TGF，这是免疫细胞的潜在负调节剂。在临床前研究中，我们表明，通过表达显性负TGF受体II（DNR），可以使EBV-CTL能够抵抗TGF的抑制作用。表达CTL的DNR保留了其效应功能，并在临床前动物模型中具有增强的抗肿瘤活性。重要的是，表达CTL的DNR没有显示出任何自主（抗原独立）CTL增殖的证据。这些结果为RAC和FDA批准的临床试验（RAC＃0507-724）提供了基本原理，其中我们正在测试（TGF）CTL在淋巴瘤患者中的安全性和有效性。 在这项研究中，我们将生成自体EBV特异性CTL，并使用表达显性阴性TGFβ受体II（DNRII）的逆转录病毒载体对其进行遗传修改，以使它们具有抗TGFββ的免疫抑制作用和嵌合抗原受体识别HER2的免疫抑制作用。将使用两个独立的向量转移基因。因此，每个细胞产品都将包含： 1）未修饰的CTL 2）HER2 CAR CTLS 3）TGF¿] DNR CTLS 4）HER2 CAR和TGF BETA DNR CTLS 我们将适当地将这些CTL转移到晚期HER2阳性肺癌患者中。我们将评估输注的安全性，并测量亚组2、3和4中基因修饰细胞的存活。我们将通过定量PCR和外周血和肿瘤组织/其他液体中的免疫染色来检测每个细胞群体（如果有）。我们预计表达TGF¿DNR的CTL种群（亚组3和4）将比缺乏这种修改的种群（亚组2）生存。我们还期望第3组和第4组的存活率等效，但是如果汽车表达本身对CTL存活产生不利影响，那么亚组3将占主导地位。另外，如果表达TGF¿DNR和CAR的细胞在肿瘤中效果更好，那么我们希望在组织样品中看到亚组4的优势。 ii。具体目标 基本的： To determine the safety of one intravenous injections of autologous Transforming Growth Factor (TGF)-resistant cytotoxic T lymphocytes (CTLs) directed to Epstein Barr virus (EBV) through their native receptor and HER2 through their chimeric antigen receptor (CAR) in patients with advanced HER2-positive lung cancers. 次要： 比较受感染的CAR-CTL的TGF耐药和非耐药成分的生存和免疫功能。 评估感染CAR-CTL的抗肿瘤作用。