Characterization and Development of a Coiled-Coil to Treat Non-Hodgkin Lymphoma

治疗非霍奇金淋巴瘤的螺旋线圈的表征和开发

• 批准号: 8526202
• 负责人: MICHAEL JACOBSEN
• 金额: $ 4.22万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8526202
• 关键词: Address; Affinity; Apoptosis; Apoptotic; B-Cell Lymphomas; B-Lymphocytes; Biocompatible; Biological; Biological Testing; CD20 Antigens; Cell Line; Cell surface; Cells; Cholecystokinin; Data; Dependence; Dependency; Development; Disease; Environment; Evaluation; Fellowship; Future; Goals; In Vitro; Inbred BALB C Mice; Induction of Apoptosis; Label; Lead; MS4A1 gene; Methods; Modeling; Molecular; Mus; Non-Hodgkin' s Lymphoma; Peptides; Polymers; Receptor Cell; Research; Research Personnel; Research Project Grants; Research Proposals; Research Training; SCID Mice; Sampling; Series; Structure; Surface; System; Testing; Therapeutic; Thermodynamics; Training; Treatment Efficacy; Water; Work; alpha helix; anticancer treatment; base; biomaterial compatibility; career; copolymer; crosslink; design; experience; immunogenic; immunogenicity; in vivo; innovation; methacrylamide; mouse model; novel; novel strategies; receptor; response; therapeutic protein; tool; treatment strategy

DESCRIPTION (provided by applicant): The central goal of this research project is to characterize and optimize an original anticancer strategy for the treatment of Non-Hodgkin Lymphoma (NHL). NHL is predominantly a B-cell lymphoma for which nearly 67,000 new cases are expected in 2011 alone. Recently, preliminary in vitro and in vivo data demonstrated a new concept for treating this disease, which is based on the assembly of coiled-coil forming peptide motifs. Here, the assembly of coiled- coils was applied to crosslink CD20 receptors on B-cell surfaces, leading to specific induction of apoptosis in these cells. The utilization of structure-specific, coiled-coil motifs as tools to control cellular phenomena is a conceptually new approach that serves as a model for future designs. In this proposal, coiled-coils-defined as specific assemblies of alpha-helix peptides-are developed as molecular apoptosis inducing agents. A heterodimeric, antiparallel coiled-coil composed of CCE and CCK peptides was previously designed and showed high structural complementarity. For application in the apoptosis induction system, the CCE peptide was covalently attached to an anti-CD20 Fab', while multiple CCK peptides were conjugated as multiple grafts to a water-soluble polymer: N-(2- hydroxypropyl) methacrylamide. Interaction of Fab'-CCE with CD20 receptors on B-cell surfaces, followed by application of Polymer-CCKn, resulted in specific coiled-coil formation (CCE+CCK) at the B- cell surfaces, leading to CD20 crosslinking and subsequent apoptosis. Proof-of-concept was demonstrated in vitro and initial in vivo studies in mice. This proposal builds on the primary data by optimization of the coiled-coil design-both its physical stability an biocompatibility. First, a series of structurally unique coiled-coils will be carefully prepared an physically characterized. Second, in vitro and in vivo evaluation of the B-cell apoptosis induction (using the new designs) will be performed, followed by evaluation of the sample immunogenicity. We hypothesize that this optimization can lead to a significant enhancement in CD20 crosslinking and B-cell apoptosis. This proposal directly targets NHL, while also providing further design and functional information for the development of future anticancer approaches.

描述（由申请人提供）：该研究项目的核心目标是表征和优化一种治疗非霍奇金淋巴瘤（NHL）的原始抗癌策略。 NHL主要是B细胞淋巴瘤，仅在2011年就预计将有近67,000例新病例。最近，初步的体外和体内数据展示了一种用于治疗这种疾病的新概念，该疾病基于组成形成肽基序的盘绕螺旋。在这里，将盘绕的线圈的组装应用于B细胞表面上的交叉链接CD20受体，从而使这些细胞中凋亡的特异性诱导。在结构特异性的，盘绕的线索图案中的利用是控制蜂窝现象的工具，是一种概念上的新方法，可作为未来设计的模型。在此提案中，盘绕的螺丝定义为α-螺旋肽的特定组件，当时开发为分子凋亡诱导剂。先前设计并显示出高结构互补性，由CCE和CCK肽组成的异二聚体，反平行线圈。为了在凋亡诱导系统中应用，将CCE肽共价附着在抗CD20 FAB上，而多个CCK肽作为多个移植物与水溶性聚合物的多个移植物结合：N-（2-羟基丙基）甲基丙烯酰胺。 Fab'-CCE与B细胞表面上的CD20受体的相互作用，然后应用聚合物-CCKN，导致在B细胞表面的特定盘绕螺旋形成（CCE+CCK），导致CD20交联和随后的凋亡。概念概念在体外证明了小鼠的体内研究和初始体内研究。该提案通过优化盘绕蛋白设计的物理稳定性的生物相容性来建立基于主要数据。首先，将仔细准备一系列结构独特的盘绕线圈。第二，B细胞凋亡诱导的体外和体内评估 （使用新设计）将进行，然后评估样品免疫原性。我们假设这种优化可以导致CD20交联和B细胞凋亡的显着增强。该建议直接针对NHL，同时还为未来的抗癌方法的开发提供了进一步的设计和功能信息。