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：
9313236
负责人：
Gerald Maxwell Cherf
金额：
$ 2.42万
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 2018-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9313236
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 Chemicals Clinic Collaborations Communities Coupled Development Disseminated Malignant Neoplasm Engineering Epitopes Evolution FDA approved Flow Cytometry Growth Hour Human In Vitro Inbred BALB C Mice Individual 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 Stream Surface Tail Techniques Testing Therapeutic Time Translating Travel United States Urokinase Urokinase Plasminogen Activator Receptor Variant Veins Yeasts cancer biomarkers cancer cell cancer therapy chemotherapy efficacy testing experimental study flexibility improved in vivo malignant breast neoplasm migration mortality mutant pressure prevent public health relevance receptor receptor function response success targeted treatment therapeutic target tool tumor tumor progression virtual

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）：癌症是美国第二常见的死亡原因，据估计，仅在2013年就导致580,350例死亡。这些死亡人数中约有90％将造成癌症发展到转移阶段，在该阶段，来自原发性肿瘤的细胞开始进入并通过血流传播，以在整个体内形成新的肿瘤。这种转移过程是由某些称为受体的蛋白质驱动的，这些蛋白质存在于癌细胞表面上，以高数量和/或升高活性相对相对到正常细胞。这些受体的丰度使癌症能够迅速生长，增殖和迁移到体内，因此许多受体已被验证为癌症治疗的治疗靶标。例如，臭名昭著的尿蛋白酶受体（UPAR）已知二十年来一直是已知的癌症靶标。 UPAR协调多个对于转移必不可少的过程；它几乎在所有癌症的水平上都存在，并且高水平与肿瘤侵袭性，对化疗，转移和总体患者生存时间的抗性增加相关。然而，尽管其重要性，但尚未获得针对该受体的FDA批准的分子。这是由于UPAR的压倒性复杂性。它具有多个功能部位，可独立地协调癌症的生长和转移。有效阻断所有功能UPAR位点的治疗性的发展仍然代表了治疗转移性癌症进展的关键障碍。在这里，我们概述了一套目的旨在克服这一障碍：目标1）使用两个先前存在的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。结果将提高对促进癌症生长和转移的关键UPAR功能的科学知识，并指出是否存在添加剂或协同作用同时定位多个功能。 AIM 2）从AIM 1改进并化学联系两个拮抗剂，以产生与超高（皮摩尔至FEMTOLOL）亲和力结合UPAR的拮抗剂，并阻止所有协调癌症进展的功能。将使用容易出错的PCR生成每个拮抗剂的107个突变体的库，在酵母表面表达，并使用高吞吐量（每秒10,000个细胞）流式细胞仪筛选与UPAR结合。这种强大的技术将选择性压力施加到在实验室基准的几个月内将数百万年的演变凝结。随后，将使用107个不同肽接头的库耦合两个具有最高亲和力的拮抗剂，并类似地筛选与UPAR的结合。具有最高亲和力的链接拮抗剂将被测试能够抑制或预防体外癌症进展的能力。 AIM 3）AIM 2的顶级候选者将对使用动物模型在体内治疗转移性癌症进行测试 - FDA需要在人类试验之前采取的步骤。我们的最终目标是继续与斯坦福大学医师科学家的合作，以开发to靶向的疗法，并将其转化为诊所以治疗转移性癌症。