Engineering a multispecific cell receptor antagonist to treat metastatic cancer

设计多特异性细胞受体拮抗剂来治疗转移性癌症

基本信息

批准号：
9114090
负责人：
Gerald Maxwell Cherf
金额：
$ 3.55万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9114090
关键词：
4T1 Affinity Amino Acids Animal Model Binding Biological Assay Biological Markers Biological Process Blood Breast Cancer Cell Cause of Death Cell Adhesion Cells Cessation of life Clinic Collaborations Communities Coupled Development Disseminated Malignant Neoplasm Engineering Epitopes Evolution FDA approved Flow Cytometry Growth Health Hour Human In Vitro Inbred BALB C Mice Injection of therapeutic agent Knowledge Length Libraries Ligands Link MDA MB 231 Malignant Neoplasms Mediating Metastatic Neoplasm to the Lung Metastatic to Monitor Mus Mutation Neoplasm Metastasis Normal Cell Oligonucleotides Patients Peptides Physicians Primary Neoplasm Process Proliferating Proteins Protocols documentation Receptor Activation Receptor Cell Resistance Scientist Site Staging Stream Surface Tail Techniques Testing Therapeutic Time Translating Travel United States Urokinase Urokinase Plasminogen Activator Receptor Variant Veins Yeasts cancer cell cancer therapy chemotherapy efficacy testing flexibility improved in vivo malignant breast neoplasm migration mortality mutant pressure prevent receptor receptor function research study response success targeted treatment therapeutic target tool tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): Cancer is the second most common cause of death in the United States, and is estimated to cause 580,350 deaths in 2013 alone. Approximately 90% of these mortalities will result from cancers progressing to the metastatic stage, where cells from the primary tumor begin to enter and travel through the blood stream to form new tumors throughout the body. This process of metastasis is driven by certain proteins called receptors that are present on the surface of cancer cells at high numbers and/or elevated activities relative to normal cells. The abundance of these receptors allows the cancer to rapidly grow, proliferate, and migrate through the body, and thus many have been validated as therapeutic targets for cancer treatment. For example, the notorious urokinase receptor (uPAR) has been a known cancer target for over two decades. uPAR coordinates multiple processes that are essential for metastasis; it is present at elevated levels in virtually all cancers, and high levels are associatd with increased tumor aggressiveness, resistance to chemotherapy, metastasis, and overall low patient survival time. However, despite its importance, no FDA- approved molecules have become available that target this receptor. This is due to the overwhelming complexity of uPAR; it has multiple functional sites that independently coordinate cancer growth and metastasis. The development of a therapeutic that efficiently blocks all functional uPAR sites still represents a critical barrier to progress in the treatment of metastatic cancer. Here, we have outlined a set of objectives aimed to overcome this barrier: Aim 1) use two pre-existing uPAR antagonists to determine the effect of simultaneously targeting the two major functional uPAR sites on cancer progression in vitro. The results will improve scientific knowledge of key uPAR functions that drive cancer growth and metastasis, and will indicate if there is an additive or synergistic effect of targeting multiple functions simultaneously. Aim 2) Improve and chemically link the two antagonists from Aim 1 to generate an antagonist that binds uPAR with ultra-high (picomolar to femtomolar) affinity and blocks all functions that coordinate cancer progression. Libraries of 107 mutants of each antagonist will be generated using error-prone PCR, expressed on the surface of yeast, and screened for binding to uPAR using high throughput (10,000 cells per second) flow cytometry. This powerful technique applies selective pressure to condense millions of years of evolution into just a few months at the lab bench. Subsequently, the two antagonists with the highest affinity will be coupled using a library of 107 different peptide linkers and similarly screened for binding to uPAR. Linked antagonists with the highest affinity will be tested for their ability to inhibit or prevent cancer progression in vitro. Aim 3) The top candidates from Aim 2 wil be tested for treating metastatic cancer in vivo using animal models - a step required by the FDA before human trials can ensue. Our ultimate objective is to continue our collaborations with physician-scientists at Stanford to develop a uPAR-targeted therapy and translate it into the clinic for treatment of metastatic cancer.

描述（由申请人提供）：癌症是美国第二大常见死因，据估计仅 2013 年就导致 580,350 人死亡。大约 90% 的死亡是由于癌症进展到转移阶段造成的，此时原发肿瘤的细胞开始进入并通过血流在全身形成新的肿瘤。这种转移过程是由某些称为受体的蛋白质驱动的，这些蛋白质以相对较高的数量和/或较高的活性存在于癌细胞表面。对正常细胞。这些受体的丰富性使得癌症能够在体内快速生长、增殖和迁移，因此许多受体已被验证为癌症治疗的治疗靶点。例如，臭名昭著的尿激酶受体（uPAR）二十多年来一直是已知的癌症靶标。 uPAR 协调对转移至关重要的多个过程；它在几乎所有癌症中都以较高的水平存在，并且高水平与肿瘤侵袭性增加、化疗耐药性、转移和患者总体生存时间短有关。然而，尽管它很重要，但目前还没有 FDA 批准的针对该受体的分子。这是由于 uPAR 极其复杂；它具有独立协调癌症生长和转移的多个功能位点。有效阻断所有功能性 uPAR 位点的治疗方法的开发仍然是转移性癌症治疗进展的关键障碍。在这里，我们列出了一组旨在克服这一障碍的目标：目标 1) 使用两种预先存在的 uPAR 拮抗剂来确定同时靶向两个主要功能性 uPAR 位点对体外癌症进展的影响。这些结果将提高对驱动癌症生长和转移的关键 uPAR 功能的科学认识，并将表明是否存在相加或协同效应同时瞄准多种功能。目标 2) 改进并化学连接目标 1 中的两种拮抗剂，生成一种拮抗剂，以超高（皮摩尔至飞摩尔）亲和力结合 uPAR，并阻断所有协调癌症进展的功能。每种拮抗剂的 107 个突变体的文库将使用易错 PCR 生成，在酵母表面表达，并使用高通量（每秒 10,000 个细胞）流式细胞术筛选与 uPAR 的结合。这项强大的技术应用选择性压力，将数百万年的进化浓缩到实验室工作台上的短短几个月内。随后，将使用 107 个不同肽接头的文库偶联具有最高亲和力的两种拮抗剂，并类似地筛选与 uPAR 的结合。具有最高亲和力的连接拮抗剂将被测试其体外抑制或预防癌症进展的能力。目标 3) 将使用动物模型对目标 2 中的顶级候选药物进行体内转移性癌症治疗的测试——这是 FDA 在进行人体试验之前所要求的步骤。我们的最终目标是继续与斯坦福大学的医师科学家合作，开发一种 uPAR 靶向疗法，并将其转化为临床治疗转移性癌症。