Localizing Immunotherapy to Improve Therapeutic Index

局部免疫治疗以提高治疗指数

基本信息

批准号：
8670703
负责人：
Karl Dane Wittrup
金额：
$ 38.55万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-04 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8670703
关键词：
Affect Affinity Antibodies Antigen Targeting Antigens Artificial nanoparticles Benchmarking Biological Assay Biomedical Engineering Biotinylation Bispecific Antibodies Blood Circulation C57BL/6 Mouse CD8B1 gene CTLA4 gene Cancer Model Carcinoembryonic Antigen Cell Surface Receptors Cells Chelating Agents Clinical Trials Collaborations Combined Modality Therapy Complex Dendritic Cells Development Diffuse Distal Docking Dose Dose-Limiting Drug Formulations Effector Cell Fc domain Freedom Genetic Engineering Genetically Engineered Mouse Human Immune Immune response Immune system Immunity Immunoglobulin G Immunosuppression Immunotherapeutic agent Immunotherapy Injection of therapeutic agent Interleukin-12 Interleukin-2 KRAS2 gene Knock-out Label Learning Ligands Liposomes MC38 Malignant Neoplasms Mediating Metals Methods Modeling Mus Myelogenous Natural Killer Cells Nature Pharmaceutical Preparations Primary Neoplasm Process Protein Engineering Proteins Protocols documentation Radioisotopes Regulatory T-Lymphocyte Research Personnel Safety Signal Transduction Site Specificity Suppressor-Effector T-Lymphocytes Surface System T-Lymphocyte TNF gene Testing Therapeutic Therapeutic Index Time Tissues Toxic effect Transgenic Mice Transgenic Organisms Tumor Antibodies Tumor Antigens Tumor Tissue Tumor-Infiltrating Lymphocytes antibody-dependent cell cytotoxicity base cancer immunotherapy cell killing cytokine design improved knowledge base lung sarcoma lymph nodes macrophage melanoma mimicry mutant nanoparticle neoplastic cell novel novel strategies public health relevance recombinase response scaffold small molecule subcutaneous tumor tumor microenvironment uptake vector

项目摘要

DESCRIPTION (provided by applicant): This new R01 proposal is a collaboration between two investigators, Wittrup and Irvine, combining protein engineering and nanoparticle synthesis expertise. Our central hypothesis is that the therapeutic index of cancer immunotherapy can be improved significantly by using novel methods to locally concentrate potent immunostimulatory molecules in tumor tissue for increased efficacy and decreased off-target toxicity. We will develop two complementary and potentially synergistic localized delivery methods for immunotherapy of cancer: pretargeting, and intratumoral nanoparticle injection. The two methods will be optimized for combined utility in syngeneic and genetically engineered mouse tumor models. We will explicate the immune therapeutic mechanisms of protocols that demonstrate efficacy. We have developed a bispecific antibody-based pretargeting protocol that provides highly tumor-specific localization of the chelator DOTA. We will site-specifically attach DOTA to the payloads IL-2, IL-12, TNF-¿, ¿-CTLA4 scFv, and ¿-CD137 scFv. These molecules were chosen due to their demonstrated immuno-therapeutic potential in clinical trials, together with significant toxicity issues. Our protocol validated for DOTA-radiometal chelates will be adapted for specific delivery of the DOTA-labeled payloads. We have devised liposomal and stabilized micellar vehicles for surface anchoring of immunostimulatory molecules, and demonstrated their efficacy and safety from intratumoral injection into B16F10 syngeneic melanoma tumors. The same bispecific antibody used for pretargeting will be anchored on the surface of these vehicles, so that the exact same DOTA-labeled payloads can be modularly tested without re-optimization of conjugation methods. The bsAb is a scaffold that enables straightforward mimicry of immunocytokines, bispecific antibodies, and Fc conjugates by noncovalent conjugation with DOTA-labeled payloads. This will enable us to benchmark safety and efficacy of our novel approaches against these more commonly used vehicles, using the same antibody for tumor targeting and identical immunostimulatory molecules. We will test these protocols in transgenic mice expressing CEA, inoculated subcutaneously with B16F10 tumors expressing human CEA. The most successful protocols will be further tested in subcutaneous MC38-CEA tumors, and then in genetically engineered KP tumors in lung and sarcoma (floxed p53 knockout and stop-floxed activated KRAS expression via Cre recombinase delivered virally.) We will closely examine the tumor microenvironment and tumor draining lymph nodes following treatment by the most efficacious protocols, for evidence of reversal of immunosuppression by Tregs, TAMs, or MDSCs. We will also test for protective immunity and antigen spreading using syngeneic tumors lacking the antigen targeted by the bsAb (CEA).

描述（由应用程序提供）：这项新的R01提案是两个研究者Wittrup和Irvine之间的合作，结合了蛋白质工程和纳米颗粒合成专业知识。我们的中心假设是，可以使用新型方法在肿瘤组织中局部浓缩有效的免疫刺激性分子来显着改善癌症免疫疗法的治疗指数，从而提高效率并降低靶向性毒性。我们将开发两种完整性和潜在的癌症免疫疗法的局部分娩方法：预先定位和肿瘤内纳米颗粒注射。这两种方法将被优化，以用于合成和基因工程的小鼠肿瘤模型中的联合实用性。我们将阐明证明有效性的方案的免疫治疗机制。我们开发了一种基于双特异性抗体的预制方案，该方案提供了螯合剂DOTA的高度肿瘤特异性定位。我们将特定地将DOTA连接到有效载荷IL-2，IL-12，TNF-¿，€-CTLA4 SCFV和€-CD137 SCFV。选择这些分子是由于它们在临床试验中表现出的免疫治疗潜力以及明显的毒性问题。我们针对DOTA-Radiometal樱桃验证的协议将适用于DOTA标记的有效载荷的特定交付。我们已经设计了脂质体和稳定的胶束车辆，用于锚定免疫刺激分子的表面，并证明了它们从肿瘤内注射到B16F10合成性黑色素瘤肿瘤中的有效性和安全性。用于预构的相同的双特异性抗体将固定在这些车辆的表面上，因此，可以在不重新审议结合方法的情况下模块化测试完全相同的DOTA标记有效载荷。 BSAB是一个支架，可以通过非共价结合与dota标记的有效载荷来直接模仿免疫细胞因子，双特异性抗体和FC结合物。这将使我们能够使用相同的抗体来靶向肿瘤和相同的免疫刺激性分子，使我们对这些更常用的车辆进行新型方法的安全性和有效性。我们将在表达CEA的转基因小鼠中测试这些方案，并用表达人CEA的B16F10肿瘤皮下接触。最成功的方案将在皮下MC38-CEA肿瘤中进一步测试，然后在肺和肉瘤中的基因工程KP肿瘤中进行（floxed p53敲除和通过CRE重物组合酶表达激活的KRAS表达，我们将实际上仔细检查肿瘤微型环境和肿瘤的效果。 Tregs，TAMS或MDSC的免疫抑制。我们还将使用缺乏BSAB（CEA）靶向的抗原的合烯肿瘤来测试保护性免疫组织化学。