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：
8476648
负责人：
Karl Dane Wittrup
金额：
$ 39.54万
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/8476648
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（由申请人证明）：这项新的R01提案是两个研究者之间的合作，将蛋白质G和Noparticle的合成专业知识结合在一起，以局部浓缩肿瘤组织中有效的免疫刺激分子，以提高效率和降低目标互补对癌症E注射的免疫疗法的潜在协同局部方法将优化用于合成的效用，并在基因工程的小鼠肿瘤模型中使用。螯合剂的定位。，� -CTLA4 SCFV和» CD137 SCFV因其在临床试验中所证明的，与DOTA-RADADIOMET CHELATES一起验证，因此选择了这些分子。 B16F10合成性黑色素瘤肿瘤。您将在皮下MC38-CEA肿瘤中进一步测试表达CEA的转基因小鼠的方案，并在肺和肉瘤中进一步测试，肺部和肉瘤中的肿瘤将进一步测试（Floxed p53敲除和D激活的KRAS kras表达）我们将通过最大的证据表明，使用BSAB（cea）靶向的抗原肿瘤（cab）（coseenial take），我们将仔细检查肿瘤微膨胀性和肿瘤排出淋巴结的淋巴结。）。