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（由申请人提供）：这项新的 R01 提案是两位研究人员 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，通过使用新方法可以显着提高癌症免疫疗法的治疗指数。我们将开发两种互补且具有潜在协同作用的癌症免疫治疗局部递送方法：预靶向和肿瘤内纳米颗粒。这两种方法将在同基因和基因工程小鼠肿瘤模型中进行优化，以阐明其免疫治疗机制，并开发出一种基于双特异性抗体的预靶向方案，该方案可提供高度肿瘤特异性的定位。我们将特定位点将 DOTA 附加到有效负载 IL-2、IL-12、TNF-¿ , ¿ -CTLA4 scFv，和 ¿ -CD137 scFv。选择这些分子是因为它们在临床试验中表现出免疫治疗潜力，并且具有显着的毒性问题。我们针对 DOTA 放射性金属螯合物验证的方案将适用于我们设计的 DOTA 标记有效负载的特定递送。用于免疫刺激分子表面锚定的脂质体和稳定胶束载体，并通过瘤内注射到 B16F10 同基因黑色素瘤肿瘤中证明了它们的有效性和安全性。用于预靶向的相同双特异性抗体将锚定在这些载体的表面上，以便可以对完全相同的 DOTA 标记的有效负载进行模块化测试，而无需重新优化缀合方法。双特异性抗体和 Fc 缀合物与 DOTA 标记的有效负载进行非共价缀合，这将使我们能够使用这些更常用的载体来衡量我们的新方法的安全性和有效性。我们将在皮下接种表达人类 CEA 的 B16F10 肿瘤的表达 CEA 的转基因小鼠中测试这些方案，最成功的方案将在皮下 MC38-CEA 肿瘤中进行进一步测试，然后在基因工程中进行测试。肺和肉瘤中的 KP 肿瘤（floxed p53 敲除并通过病毒传递的 Cre 重组酶停止 floxed 激活的 KRAS 表达。）我们将仔细检查肿瘤通过最有效的方案治疗后的微环境和肿瘤引流淋巴结，以获取 Tregs、TAM 或 MDSC 逆转免疫抑制的证据。我们还将使用缺乏 bsAb（CEA）靶向抗原的同源肿瘤来测试保护性免疫和抗原扩散。）。