Abrogating human CD59 activity for antibody-based cancer therapy

消除人类 CD59 活性以进行基于抗体的癌症治疗

基本信息

批准号：
8009892
负责人：
Xuebin Qin
金额：
$ 21.46万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8009892
关键词：
Adopted Adverse effects Affinity Algorithms Amino Acids Animals Antibodies B-Cell Lymphomas B-Cell NonHodgkins Lymphoma Binding Biological Assay Body Weight CD46 Antigen CD55 Antigens CDC2 gene Caliber Cell Line Cell membrane Cells Classical Complement Pathway Complement Complement Membrane Attack Complex Complement component C8 Complement-Dependent Cytotoxicity Computer Simulation Culture Media Cytolysis Development Dose Drug Kinetics Endotoxins Epitopes Erythrocytes Escherichia coli Exposure to FBXW7 gene Follicular Lymphoma Fostering Future Hemolysis Human Immune response Immunotherapy In Vitro Indolent Inhibitory Concentration 50 Length Lymphoma Major Histocompatibility Complex Malignant Neoplasms Measurement Mediating Mediator of activation protein Membrane Monitor Nature Normal Cell Nude Mice Peptides Phenotype Reagent Recombinant Proteins Recombinants Regimen Research Resistance Specificity Streptococcus intermedius Surgical Replantation T cell response T-Cell Proliferation T-Cell Receptor Testing Therapeutic Therapeutic antibodies Toxic effect Toxin Transgenic Mice Transgenic Organisms Up-Regulation Work Xenograft procedure antibody-dependent cell cytotoxicity base cancer cell cancer therapy cell killing design immunogenicity implantation in vivo in vivo Model inhibitor/antagonist innovation intermedilysin neoplastic cell novel novel strategies peptidomimetics polymerization prevent public health relevance receptor rituximab success tumor growth tumor xenograft

项目摘要

DESCRIPTION (provided by applicant): CD59 is a key regulator for restricting the formation of the membrane attack complex (MAC) of complement by binding to complement components C8 and C9 and preventing C9 incorporation and polymerization in the cell membrane. Complement is a key mediator for antibody-based cancer therapy such as complement-dependent cytotoxicity (CDC) and antibody-dependent-cell mediated cytotoxicity (ADCC). CD59 is universally expressed in all cells and highly expressed in many kinds of cancer cells including B-cell lymphoma. CD59 appears to be the membrane complement regulator that is most effective at protecting tumor cells from antibody-mediated CDC. Extensive studies indicate that upregulation of CD59 is responsible for lymphoma resistance to rituximab treatment. Therefore, it is imperative for us to develop a molecule capable of abrogating CD59 function in cancer cells and facilitating antibody-mediated cancer therapy. However, targeted toxicity effect from miniantibody specific against hCD59 and less efficacy of C8 or C9 peptides limit them for therapeutic purposes. Intermedilysin (ILY), a cytolytic pore-forming toxin secreted by Streptococcus intermedius, lyses only human cells due to its receptor specificity for human CD59 (hCD59). Recently, we used hCD59 transgenic mice to confirm the finding that ILY binds only to hCD59 in vivo. Domain 4 of ILY binds to AA42-58 in hCD59, which also participate in the binding to C8 and C9. Thus, we hypothesize that the truncated ILY presenting only domain 4 will specifically abrogate hCD59 function and facilitate antibody- mediated and complement-dependant cancer cytolysis. Consistently, our preliminary results have demonstrated that the recombinant protein (114AA) derived from the ILY domain 4 (rILYd4) specifically blocks hCD59 function ex vivo. Furthermore, we demonstrated that application of rILYd4 (IC50 = 33 nM in vitro and effective dose = 2.5 ug/g body weight in vivo ) to B lymphoma cell lines (RAMOS), which are resistant to Rituximab-mediated CDC (so called RRR cells), sensitized them to Rituximab-mediated CDC in lymphoma without off target toxicity effects. We propose to use rituximab as a therapeutic antibody in lymphoma xenografted athymic nude mice that express transgenic hCD59 to test this hypothesis. Specifically, we will determine and the efficacy and specificity of rILY4 (specific aim 1) and characterize rILYd4 immunogenicity (Aim 2). Success in this application will yield a newly-identified CD59 inhibitor, domain 4 of ILY as an adjunct to not only rituximab-based lymphoma therapy, but also other antibody-based cancer therapy. PUBLIC HEALTH RELEVANCE: In this application, we will assess the in vivo efficacy of the nature product rILYd4 for antibody (rituximab)-mediated cancer (B-lymphoma) therapy. Success of this work will foster us to develop novel anti-cancer reagents, an anti-human CD59 inhibitor rILYd4, which can significantly facilitate antibody- mediated cancer therapy, not only in lymphoma, but also in other cancers.

描述（由申请人提供）：CD59是通过与补体组件C8和C9结合，并防止在细胞膜中结合和聚合来限制补体组件C8和C9的膜形成补体的形成（MAC）。补体是基于抗体癌症治疗的关键介体，例如补体依赖性细胞毒性（CDC）和抗体依赖性细胞介导的细胞毒性（ADCC）。 CD59在所有细胞中普遍表达，并且在包括B细胞淋巴瘤在内的许多类型的癌细胞中高度表达。 CD59似乎是最有效保护肿瘤细胞免受抗体介导的CDC的膜补体调节剂。广泛的研究表明，CD59的上调负责淋巴瘤对利妥昔单抗治疗的抗性。因此，我们必须开发能够在癌细胞中废除CD59功能并促进抗体介导的癌症治疗的分子。但是，针对HCD59的微抗体特异性的靶向毒性作用以及C8或C9肽的疗效限制了它们出于治疗目的。 Intermedilysin（Ily）是一种由Intermedius链球菌分泌的胞溶毛孔形成毒素，由于其受体特异性对人CD59（HCD59）而仅裂解人类细胞。最近，我们使用HCD59转基因小鼠来确认ILY在体内仅与HCD59结合的发现。 ILY的域4与HCD59中的AA42-58结合，这也参与了与C8和C9的结合。因此，我们假设仅截断的ILY仅呈现域4将特别废除HCD59功能，并促进抗体介导的和补体依赖性依赖性癌细胞溶解。一致地，我们的初步结果表明，重组蛋白（114AA）衍生自ILY结构域4（Rilyd4）特异性阻断了HCD59功能在体内。此外，我们证明了Rilyd4（IC50 = 33 nm的体外和有效剂量= 2.5 ug/g体重体重）在B淋巴瘤细胞系（RAMOS）（RAMOS）中，对利妥昔单抗介导的CDC（所谓的RRR细胞）抗性，对ritiTuximab介导的cDC构成质量效应，对降低了质量cDC的效果。我们建议将利妥昔单抗作为一种表达转基因HCD59的淋巴瘤异常裸鼠的治疗抗体来检验该假设。具体而言，我们将确定RILY4（特定目标1）的功效和特异性并表征Rilyd4免疫原性（AIM 2）。在此应用中的成功将产生新鉴定的CD59抑制剂，即ILY的域4，不仅是基于利妥昔单抗的淋巴瘤疗法的辅助手段，而且是其他基于抗体的癌症治疗。公共卫生相关性：在此应用中，我们将评估自然产物Rilyd4对抗体（利妥昔单抗）介导的癌症（B-淋巴瘤）疗法的体内功效。这项工作的成功将促进我们开发新型抗癌试剂，即一种抗人CD59抑制剂Rilyd4，不仅在淋巴瘤中，而且在其他癌症中也可以显着促进抗体介导的癌症治疗。