Targeting EMP2 for the treatment of triple negative breast cancer with novel anti-EMP2 Granzyme B immunoconjugates

使用新型抗 EMP2 颗粒酶 B 免疫缀合物靶向 EMP2 治疗三阴性乳腺癌

基本信息

批准号：
9810531
负责人：
Khalid A Mohamedali
金额：
$ 21.84万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9810531
关键词：
Affinity Animals Antibodies Apoptosis Binding Biodistribution Biological Markers Breast Cancer Model Breast Cancer therapy Cancer Diagnostics Cause of Death Cell Death Cell Death Process Cell Proliferation Cell surface Cells Chimeric Proteins Clinic Cytoplasm Cytotoxic agent Data Diagnosis Disease Dose Drug Kinetics Endometrial Engineering Epithelial Evaluation Female Genital Neoplasms Generations Goals Granzyme Growth Factor Half-Life Human IgG1 Immunoconjugates Immunoglobulin G Immunotoxins In Vitro Individual Kinetics Lead Length Link Magic Malignant Neoplasms Maximum Tolerated Dose Membrane Proteins Modeling Molecular Monoclonal Antibodies Mus Neoplasm Metastasis Normal tissue morphology Ovarian Pattern Positioning Attribute Primary Neoplasm Production Protein Engineering Proteins Reagent Recombinant Antibody Recombinants Research Resistance Science Serine Protease Specificity Testing Therapeutic Therapeutic Index Time TimeLine Toxic effect Toxin Tumor Burden University of Texas M D Anderson Cancer Center Woman Work Xenograft procedure alpha Toxin base cancer therapy chemotherapy comparative efficacy cytotoxic cytotoxicity design dosage drug candidate efficacy testing glycosylation immunogenicity improved in vivo indexing malignant breast neoplasm mammary epithelium neoplastic cell new therapeutic target novel novel therapeutics pre-clinical preclinical study protein expression research clinical testing targeted treatment triple-negative invasive breast carcinoma tumor

项目摘要

PROJECT SUMMARY/ABSTRACT Immunoconjugates are designed to focus the delivery of highly cytotoxic agents to specific target cells using monoclonal antibodies, potentially improving the therapeutic index of the agent. To this end, we propose to deliver a cytotoxic protein payload with an anti-EMP2 IgG1 antibody. EMP2, or epithelial membrane protein-2, is a biomarker that is highly expressed in the majority (63%; n=97) of invasive breast cancer tumors examined compared to healthy mammary epithelium, and high EMP2 expression is observed in over 75% of triple negative breast cancer cases examined where EMP2 is observed in both the primary tumor as well as in metastatic lesions. As engineered recombinant antibodies hold great promise for cancer diagnostics and therapy, we have carefully assembled a research team centered at UCLA and The University of Texas MD Anderson Cancer Center to develop recombinant anti-EMP2 antibodies fused to the human serine protease granzyme B for therapy. The group at UCLA has previously shown that a full length IgG1 against EMP2 promotes apoptosis both in vitro and in vivo in a number of EMP2 positive tumors, but as new data suggests that the anti-EMP2 IgG1 can internalize rapidly, we hypothesize that it may serve as a novel candidate for drug conjugation. The use of Granzyme B (GrB) as an immunoconjugate has been proposed as a “magic bullet” as, once delivered to the cytoplasm of a cell, it activates apoptosis pathways with little to no induction of immunogenicity. The Mohamedali lab has utilized human GrB as an effective payload for the generation of recombinant cell death-inducing fusion proteins and has clearly demonstrated that GrB-containing fusion constructs have impressive and highly selective cytotoxic effects when delivered to the cytoplasm by either antibody or growth factor cell targeting carriers. In this proposal, with the explicit goal of advancing the science of anti-EMP2 antibodies as well as GrB as a cytotoxic payload for clinical testing, we will create and determine the efficacy of two anti-EMP2 IgG1- GrB conjugates. Accordingly, the specific aims propose to design and test the efficacy of GrB linked to an anti-EMP2 IgG1 for cytotoxicity as well as start creating a preclinical package to understand its functional affinity and stability. As the naked antibody cross-reacts between human and mouse, the toxicity profile of the immunoconjugate, including pharmacokinetics and maximum tolerated dosage, will also be determined. Given its high expression in a number of gynecological tumors including ovarian and endometrial, these studies will be important to position EMP2 as a viable target for cancers in women. It will also, given the need for new toxins with low immunogenicity, help position granzyme B in the forefront of antibody linked toxins.

项目摘要/摘要免疫共轭物的设计目的是将高度细胞毒性剂的递送到特定的靶细胞中单克隆抗体，有可能改善药剂的治疗指数。为此，我们建议用抗EMP2 IgG1抗体提供细胞毒性蛋白有效载荷。 EMP2或上皮膜蛋白2，是一种在大多数（63％; n = 97）的浸润性乳腺癌肿瘤中高度表达的生物标志物与健康的乳腺上皮相比，在超过75％的三重阴性中观察到高EMP2表达乳腺癌病例检查了在原发性肿瘤和转移性中观察到EMP2的位置病变。由于工程重组抗体对癌症诊断和疗法具有很大的希望，因此小心地组装了一个以加州大学洛杉矶分校和德克萨斯大学MD Anderson Cancer为中心的研究团队开发与人丝氨酸蛋白酶颗粒B融合的重组抗EMP2抗体的中心治疗。加州大学洛杉矶分校的组以前表明，针对EMP2的全长IgG1促进了凋亡在许多EMP2阳性肿瘤中的体外和体内都有体内，但由于新数据表明抗EMP2 IgG1 可以迅速内化，我们假设它可以作为药物结合的新候选人。使用 Granzyme B（GRB）作为免疫缀合物被提议作为“魔术子弹”，因为一旦送达细胞的细胞质，它激活凋亡途径，几乎没有免疫原性。穆罕默德实验室已将人类GRB用作生成重组细胞死亡诱导融合的有效有效载荷蛋白质并清楚地表明，含GRB的融合构建体具有令人印象深刻且高度当通过抗体或生长因子细胞靶向传递到细胞质时，选择性细胞毒性作用载体。在此提案中，明确的目标是推进抗EMP2抗体的科学以及GRB的科学细胞毒性有效载荷用于临床测试，我们将创建并确定两个抗EMP2 IgG1-GRB的效率共轭。彼此之间，具体的目的是设计和测试与抗EMP2相关的GRB的效率 IgG1用于细胞毒性，并开始创建一个临床前包，以了解其功能亲和力和稳定。随着人与小鼠之间的裸抗体交叉反应，还将确定免疫缀合物，包括药代动力学和最大耐受剂量。给出它在许多妇科肿瘤（包括卵巢和子宫内膜）中的高表达，这些研究将是将EMP2定位为女性癌症的可行靶标。鉴于需要新的毒素具有低免疫原性，有助于将颗粒状B位置在连接的毒素抗体的最前沿。