Targeting Effector Immune cells to Cancer with Chemically Self-Assembled Nanorings (CSANs)

使用化学自组装纳米环 (CSAN) 将效应免疫细胞靶向癌症

基本信息

批准号：
10600820
负责人：
CARSTON R. WAGNER
金额：
$ 55.91万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10600820
关键词：
Affinity Antibiotics Antibodies Antigen Targeting Antigens Avidity B lymphoid malignancy Binding Bispecific Antibodies Bite Breast Cancer Cell CD3 Antigens Cancer Model Cancer Patient Cell Adhesion Molecules Cell membrane Cell surface Cells Chemicals Chimeric Proteins Clinical Consumption Continuous Infusion Development Dihydrofolate Reductase Dose Effector Cell Engineering Epithelial Cells FDA approved Genetic Engineering Heterogeneity Immune Immunotherapy In Vitro Incubated Inflammatory Libraries Ligands Malignant Neoplasms Methodology Methods Methotrexate Molecular Mus Natural Killer Cells Normal tissue morphology Oligonucleotides Patients Pharmaceutical Preparations Primary Neoplasm Progression-Free Survivals Proliferating Prosthesis Resistance Resistance development Safety Solid Neoplasm Specificity Surface Antigens System T cell therapy T-Cell Activation T-Cell Receptor T-Lymphocyte Testing Time Toxic effect Trimethoprim Tumor Antigens Tumor Markers Tumor-infiltrating immune cells arm bi-specific T cell engager cancer cell cancer immunotherapy cancer stem cell cancer therapy chemical addition chimeric antigen receptor chimeric antigen receptor T cells clinical development clinically relevant combat cost effective crosslink cytokine cytotoxicity fluorophore genetic approach in vivo method development neoplastic cell non-genetic patient derived xenograft model pharmacologic pre-clinical radiotracer self assembly success triple-negative invasive breast carcinoma trispecific killer engager tumor tumor eradication tumor initiation

项目摘要

Of the many immunotherapy approaches under development, the ability to use bispecific antibodies or chimeric antigen receptors (CARs) to direct T-cells to selectively kill tumor cells has demonstrated significant early success. Typically, bispecific antibodies or bispecific T-cell engagers (i.e., BiTes) cross-link T-cells by binding to CD3 and to a target cancer cell surface antigen, usually through a monovalent interaction. An alternative approach is to genetically engineer a cancer patient’s T-cells to express a single chain antibody (scFv)-CD3ζ fusion protein that can target the cancer cell surface antigen. After re-introduction into the patient, CAR-expressing T-cells have been able to selectively eliminate the target cancer cells. While successful, the genetic engineering of cell surfaces is time consuming and irreversible and the use of BiTes requires continuous infusion. Furthermore, the resistance to both approaches due to antigen loss has been observed. Our group has shown that two dihydrofolate reductase molecules (DHFR2) fused to an αCD3 single chain antibody (scFv) can be engineered to spontaneously self-assemble upon the addition of the chemical dimerizer, bis-methotrexate (BisMTX), into either highly stable octavalent chemically self-assembled nanorings (CSANs). CSANs have been prepared with BisMTX containing a third arm, thus enabling it to be conjugated to oligonucleotides, fluorophores, radiolabels and drugs. Recently, we have prepared αEpCAM/αCD3 CSANs and αCD133/αCD3-CSANs. The bispecific CSANs rapidly (min) and stably (days) bind to CD3 on T-cell membranes, thus forming chemically self assembled prosthetic antigen receptor (PAR) T-cells. Upon incubation of the PAR T-cells with EpCAM+ and/or CD133+ cancer cells, rapid and selective killing of primary and tumor initiating cancer stem cells (CSC) was observed. We have also demonstrated with an orthotopic murine cancer model that αEpCAM and αCD133 PAR T-cells are non-toxic and able in combination to eradicate tumors in vivo. A unique safety feature of our approach is the ability to remove the CSANs from the T-cells by dosing with the FDA-approved non-toxic antibiotic trimethoprim at clinically relevant concentrations, thus allowing us to deactivate the cells pharmacologically and reduce cytokine release. Consequently, as an alternative to current approaches, we determine the generality T-cell induced killing and eradication of TNBC tumors with αEpCAM/α-CD3-CSANS and αCD133/αCD3-CSANS. In addition, we will develop a tripspecific αEpCAM/αCD133/αCD3 CSANs that will allow the simultaneous elimination of TNBC primary tumor cells and CSC. The successful completion of the project milestones should result in the elucidation of the key features governing a chemical biologically based non-genetic and reversible method for T-cell targeting, as well as the importance of CD133 on TNBC proliferation. These rules will be applicable to the clinical development of anti-cancer immunotherapy with greater selectivity, lower toxicity and a reduced ability for the development of resistance.

在许多正在开发的免疫治疗方法中，使用双特异性抗体或嵌合抗原受体（CAR）指导 T 细胞选择性杀死肿瘤细胞已被显着证明通常，双特异性抗体或双特异性 T 细胞接合剂（即 BiTes）通过以下方式交联 T 细胞。通常通过单价相互作用与 CD3 和靶癌细胞表面抗原结合。另一种方法是对癌症患者的 T 细胞进行基因改造，使其表达单链抗体 (scFv)-CD3ze融合蛋白可以靶向癌细胞表面抗原，重新引入患者体内后，表达 CAR 的 T 细胞能够选择性地消除目标癌细胞。细胞表面的基因工程耗时且不可逆，使用 BiTes 需要此外，还观察到由于抗原丢失而对两种方法产生耐药性。我们的小组已经证明两个二氢叶酸还原酶分子（DHFR2）融合到αCD3单链上抗体（scFv）可以被设计为在添加化学物质后自发自组装二聚体，双甲氨蝶呤 (BisMTX)，形成高度稳定的八价化学自组装纳米环 (CSAN) 是用含有第三臂的 BisMTX 制备的，从而使其能够与最近，我们制备了αEpCAM/αCD3 CSAN。和 αCD133/αCD3-CSAN 双特异性 CSAN 快速（分钟）且稳定（天）与 T 细胞上的 CD3 结合。膜，从而形成化学自组装的假体抗原受体（PAR）T 细胞。将 PAR T 细胞与 EpCAM+ 和/或 CD133+ 癌细胞一起孵育，快速选择性地杀死原代细胞我们还通过原位观察到了肿瘤起始癌症干细胞（CSC）。小鼠癌症模型表明 αEpCAM 和 αCD133 PAR T 细胞无毒并且能够联合使用我们的方法的一个独特的安全特征是能够从体内去除 CSAN。 T 细胞通过 FDA 批准的无毒抗生素甲氧苄啶按临床相关浓度给药，从而使我们能够通过药理学使细胞失活并减少细胞因子的释放。经过检查，作为当前方法的替代方案，我们确定了 T 细胞诱导杀伤的通用性并用 αEpCAM/α-CD3-CSAN 和 αCD133/αCD3-CSAN 根除 TNBC 肿瘤。开发旅行特异性 αEpCAM/αCD133/αCD3 CSAN，以同时消除 TNBC 原代肿瘤细胞和 CSC 项目里程碑的成功完成应导致阐明基于化学生物学的非遗传和可逆方法的关键特征 T 细胞靶向以及 CD133 对 TNBC 增殖的重要性这些规则将适用于。选择性更强、毒性更低、治疗效果更佳的抗癌免疫疗法的临床开发发展抵抗力的能力。