Synthetic Lethal Targeting of Growth Factor Receptors

生长因子受体的合成致死靶向

基本信息

批准号：
9218305
负责人：
BLAKE PETERSON
金额：
$ 32.91万
依托单位：
UNIVERSITY OF KANSAS LAWRENCE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-02-01 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9218305
关键词：
Affect Alpha Cell Antibodies Antibody-drug conjugates Antineoplastic Agents BRCA1 gene Binding Biochemical Pathway Biological Assay Breast Cancer Cell Cancer Model Cancer cell line Cell Culture Techniques Cell Death Cell Surface Proteins Cell Surface Receptors Cell surface Cells Cellular Membrane Circular Dichroism Spectroscopy Collaborations Computer Simulation Cytoplasm Cytotoxin Data Development Disseminated Malignant Neoplasm Disulfides Drug Delivery Systems Drug Targeting Drug resistance ERBB2 gene ERBB3 gene Early Endosome Endocytosis Endosomes Epidermal Growth Factor Receptor Evaluation FDA approved Future Genes Genetic Glutathione Growth Factor Growth Factor Overexpression Growth Factor Receptors Health Heterodimerization Homo Human Immunotoxins In Vitro Individual Lethal Genes Link Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of pancreas Masks Mediating Membrane Modeling Molecular Molecular Conformation Molecular Mechanisms of Action Monoclonal Antibodies Mutation NMR Spectroscopy NOD/SCID mouse Normal Cell Patients Peptides Proteins Research Research Personnel Signal Pathway Skin Structure Structure-Activity Relationship Therapeutic Toxic effect Toxin Trastuzumab Tubulin Binding Agent United States National Institutes of Health Validation Virulent Xenograft Model antibody conjugate base biophysical techniques cancer cell cancer therapy cell type cytotoxic cytotoxicity design differential expression disulfide bond efficacy evaluation endosome membrane fluorophore improved in vivo innovation killings malignant breast neoplasm mouse model novel novel strategies novel therapeutic intervention peptide structure pre-clinical preclinical study prevent receptor seal self assembly small molecule synergism uptake

项目摘要

One of the greatest challenges facing the development of improved cancer therapeutics is the need to selectively kill all of the cancer cells in a patient without harming normal cells. To achieve this high level of selectivity, the genetic concept of synthetic lethality offers a promising strategy. This concept is based on the observation that mutations in two different genes that both contribute an essential biochemical pathway, such as the genes BRCA1/2 and PARP, can be exploited to make certain cancers uniquely sensitive to anticancer agents. In this mechanism, disruption of either gene alone does not affect cellular viability, but agents or mutations that affect both genes are lethal. Based on our preliminary research results, we propose here to extend the concept of synthetic lethality to the targeting of pairs of growth factor receptors that drive the proliferation of highly aggressive cancers. To accomplish this objective, we will create novel antibody conjugates, similar in structure to the FDA-approved anticancer antibodies Kadcyla and Adcetris, designed with the unique ability to synergistically kill cancer cells that express two distinct cell surface receptors. To enable this cytotoxic synergy, one targeting antibody will be linked via a disulfide to a cell-impermeable cytotoxin that is incapable of unaided passage across cellular membranes. When this first antibody binds a specific growth factor receptor on the cell surface, and is internalized by endocytosis, the stability of the disulfide, in conjunction with the cell-impermeability of the cytotoxin, will cause entrapment in membrane- sealed endosomes. This entrapment will prevent toxicity unless membranes of these endosomes are disrupted by co-administration with a secondary agent. To synergistically kill cancer cells, this antibody conjugate will be co-administered with a second anti-growth factor antibody linked to a non-toxic endosome disruptive peptide. Release of this peptide from the second antibody will form pores in endosomal membranes. These pores will enable cytosolic glutathione to enter endosomes, break the disulfide bond linking the toxin to the first antibody, and activate toxicity by enabling escape of the cytotoxin into the cytoplasm. This unique design of antibody conjugates will provide high selectivity for killing specific cancer cells that express two distinct cell surface receptors without affecting normal cells that express one of these two target proteins. This novel approach, termed here synthetic lethal targeting, will be pursued by the synthesis of masked cytotoxins and endosome disruptive peptides, the elucidation of mechanisms of pore formation in endosomal membranes, conjugation of these agents to antibodies that bind the growth factor receptors EGFR, HER2, and HER3, and evaluation of efficacy in vitro and in mouse models of cancer. Given that co-expression of HER2/EGFR and HER2/HER3 in breast cancer, and EGFR/HER3 in lung and pancreatic cancer, drives the proliferation of some of the most incurable cancers, extending synthetic lethality by targeting two distinct growth factor receptors could provide a less toxic platform to eradicate cancer across cell types.

改善癌症治疗剂的发展面临的最大挑战之一是需要选择性地杀死患者中的所有癌细胞，而不会损害正常细胞。达到这一高水平选择性，合成致死性的遗传概念提供了有前途的策略。这个概念是基于观察到两个不同基因的突变都促进了必不可少的生化途径，由于基因BRCA1/2和PARP可以被利用以使某些癌症对抗癌的独特敏感代理商。在这种机制中，仅两个基因的破坏都不会影响细胞活力，而是影响药剂或影响这两个基因的突变都是致命的。根据我们的初步研究结果，我们在这里建议将合成致死性的概念扩展到驱动驱动的生长因子受体对的靶向高度侵略性癌症的扩散。为了实现这一目标，我们将创建新颖的抗体缀合物，与FDA批准的抗癌抗体Kadcyla和Adcetris相似具有协同杀死表达两个不同细胞表面受体的癌细胞的独特能力。到启用这种细胞毒性协同作用，一种靶向抗体将通过二硫化物连接到可渗透的细胞细胞毒素无能力在细胞膜上无助。当第一种抗体结合A时细胞表面上的特定生长因子受体，并通过内吞作用内部化，稳定性二硫化物与细胞毒素的细胞覆盖能力结合，将导致膜中陷入困境密封的内体。除非这些内体的膜被干扰，否则这种陷阱将防止毒性通过与次要代理共同管理。为了协同杀死癌细胞，这种抗体结合物将是与与无毒内体颠覆性肽相关的第二抗生长因子抗体共同管理。从第二抗体中释放该肽将在内体膜中形成孔。这些毛孔会使胞质谷胱甘肽能够进入内体，打破将毒素与第一抗体联系起来的二硫键，并通过使细胞毒素逃脱到细胞质中来激活毒性。这种独特的抗体设计缀合物将为杀死表达两个不同细胞表面的特定癌细胞提供高选择性受体不影响正常细胞，这些细胞表达这两种靶蛋白之一。这种新颖的方法，在这里称为合成致死靶标，将通过掩盖细胞毒素和内体的合成来追求破坏性肽，内体膜中孔形成机制的阐明，结合这些试剂与结合生长因子受体EGFR，HER2和HER3的抗体，并评估体外和小鼠模型的功效。鉴于HER2/EGFR和HER2/HER3的共表达乳腺癌和肺癌和胰腺癌中的EGFR/HER3驱动了一些最多的人无法治愈的癌症，通过靶向两个不同的生长因子受体来扩展合成致死性，可以提供消除细胞类型的癌症的毒性平台较小。