VLR-CART Cancer Immunotherapy

描述（由申请方提供）：Novab的使命是利用七鳃鳗产生的可变淋巴细胞受体（VLR）的独特性质，开发有效的、互补的和/或上级的治疗药物，这些治疗药物目前来自哺乳动物单克隆抗体（mMAb）。在本申请中，我们寻求启动并证明该使命内的一个商业项目的概念验证，该商业项目是将VLR应用于嵌合抗原受体（CAR）的设计，嵌合抗原受体（CAR）目前用作遗传工程化T细胞免疫疗法的肿瘤靶向组分。我们的假设是，由于独立进化的免疫球蛋白（Ig）的有颌脊椎动物，七鳃鳗VLRs提出了对治疗细胞的目标将识别表位是独特的，从那些使用标准的哺乳动物免疫和单克隆抗体（即IG）生产。因此，VLR代表了用于鉴定癌细胞靶标和产生肿瘤细胞的新平台。 改进的诊断和生物治疗（例如单克隆VLR或VLR-CAR）。最近，表达汽车的T细胞（CART）已经在有限的一组其他难治性癌症中表现出高度的功效。目前，CAR多样性受到有效MAb结合结构域的鉴定和可用性的限制，这反过来不仅受到哺乳动物遗传多样性的限制，而且受到IG结构几何形状和对大多数人源抗原的预先存在的免疫耐受屏障的限制。我们预测，VLR技术将允许更特异的癌细胞靶向，并明显扩大可以使用CART有效治疗的癌症类型的范围。具体来说，我们将使用两个独立的造血肿瘤模型来测试这一预测。使用高通量筛选和分子克隆技术，针对小鼠BCL肿瘤细胞系和原代人多发性骨髓瘤细胞产生肿瘤特异性VLR。这些VLR将用作新型汽车的抗原结合组分。我们将使用一系列体外测试（里程碑1）和免疫活性和免疫受损小鼠（里程碑2）证明汽车是否可以用作可行的抗肿瘤治疗。本提案中提出的里程碑的完成将提供基本的概念验证证据，证明VLR-CART是开发有效抗癌生物治疗药物的科学可行的平台。

DESCRIPTION (provided by applicant): The mission of Novab is to utilize the unique properties of lamprey generated Variable Lymphocyte Receptors (VLRs) to develop effective, complementary and/or superior therapeutics to those currently derived from mammalian monoclonal antibodies (mMAbs). In the current application, we seek to initiate and demonstrate proof of concept for one commercial project within this mission, which is the application of VLRs to the design of chimeric antigen receptors (CARs) that currently are used as the tumor targeting component of genetically- engineered T cell immunotherapies. Our hypothesis is that due to independent evolution from the immunoglobulins (Igs) of jawed vertebrates, lamprey VLRs raised against therapeutic cell targets will recognize epitopes that are unique from those obtained using standard mammalian immunization and MAb (i.e. Ig) production. Therefore, VLRs represent a novel platform for the identification of cancer cell targets and the generation of improved diagnostics and biotherapeutics (e.g. monoclonal VLRs or VLR-CARs). Recently, CARs expressing T cells (CARTs) have demonstrated a high degree of efficacy in a limited set of otherwise refractory cancers. Currently, CAR diversity is restricted by the identification and availability of effective MAb binding domains, which in turn is restricted by not only mammalian genetic diversity but also by Ig structural geometry and pre-existing immune tolerance barriers to most antigens of human origin. We predict that VLR technology will allow for more specific cancer cell targeting and appreciably expand the range of cancer types that can be effectively treated using CARTs. Specifically, we will test this prediction using two independent hematopoietic tumor models. Using high-throughput screening and molecular cloning technology, tumor- specific VLRs were generated against a mouse BCL tumor cell line and primary human multiple myeloma cells. These VLRs will be utilized as the antigen binding components of novel CARs. We will demonstrate if VLR- CARs can be used as viable anti-tumor treatments using a series of in vitro tests (Milestone 1) and immune- competent and immune-compromised mice (Milestone 2). Completion of the Milestones presented in this proposal will provide the fundamental proof of concept evidence that VLR-CARTs are a scientifically-viable platform for the development of effective anti-cancer biotherapeutics.