Development of a Coiled-Coil therapeutic for Non-Hodgkin's Lymphoma

开发用于非霍奇金淋巴瘤的螺旋线圈疗法

基本信息

批准号：
8933950
负责人：
Jonathan M Hartley
金额：
$ 3.33万
依托单位：
UNIVERSITY OF UTAH
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：
2014-06-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8933950
关键词：
Adverse effects Amino Acids Analytical Chemistry Animal Experiments Animal Model Antibodies Apoptosis Architecture B-Cell Lymphomas B-Lymphocytes Binding Biocompatible Biocompatible Materials Biological Biological Assay CD20 Antigens Case Study Cell Death Cell Line Cell surface Cells Cellular Structures Charge Chemistry Circular Dichroism Complex Data Development Diagnosis Disease Energy Transfer Evaluation Fab Immunoglobulins Goals Image Image Analysis Imaging Techniques Immunoglobulin Constant Region In Vitro Induction of Apoptosis Institutes Lead Length Link MS4A1 gene Medical Membrane Proteins Methods Modification Molecular Weight Monoclonal Antibodies Multiple Sclerosis Non-Hodgkin&apos s Lymphoma Patients Peptides Pharmaceutical Preparations Polymer Chemistry Polymers Progressive Multifocal Leukoencephalopathy Proteins Reporting Research Personnel Resolution Rheumatoid Arthritis Roche brand of rituximab Rodent Signal Transduction Structure System Techniques Testing Therapeutic Therapeutic Clinical Trial Training United States Vertebral column Work cancer cell cancer therapy copolymer crosslink density design experience functional group improved in vivo insight lung injury methacrylamide nanomaterials novel novel strategies novel therapeutics optical imaging polymerization professor protein crosslink public health relevance quantitative imaging response rituximab self assembly single molecule submicron tool

项目摘要

DESCRIPTION (provided by applicant): The goal of this proposal is to develop a new therapeutic system to improve the treatment of non-Hodgkin's lymphoma (NHL). Cases of NHL have doubled since 1980 and there are an estimated 69,000 new diagnoses in 2013. B-cell lymphomas constitute 85% of NHL cases; therefore, the new anti-cancer treatment relies on the cross-linking of CD20 receptors on the B-cell surface to induce apoptosis. The therapeutic system has already been shown to induce apoptosis of B-cells in vitro and in vivo. To achieve the goal of the project, we will optimize the therapeutic system for the treatment of NHL. The therapeutic system utilizes a pair of oppositely charged peptides that form an antiparallel coiled coil. The coiled coil peptides (CC1/CC2) form physical crosslinks through biorecognition. Biorecognition leads to crosslinking of CD20 receptors on B-cells. A CD20-specific Fab' fragment is attached to one of the coiled coil peptides (Fab'- CC1), and multiple copies of the complementary peptide are attached to a N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer (P-CC2; P is the polymer backbone). We hypothesize that increasing the crosslinking of CD20 through conjugate structure modifications of our previous design will result in increased apoptosis. To test our hypothesis and accomplish our goals, we propose to design and synthesize conjugates of varying architecture and coiled coil peptides containing L- and/or D-amino acid residues. These conjugates will be highly tunable, and can self-assemble into highly ordered structures due to specific interactions between peptides. The conjugate biorecognition will be evaluated and design parameters will be identified that enhance biorecognition. Structural features to optimize include: Mw of HPMA backbone, spacing of peptide grafts, and spacer length between peptides and polymer/Fab'. These studies will provide valuable insight into specific design features that improve self-assembly of coiled coil systems in general. Self-assembly of the conjugates at the cell surface and the crosslinking of CD20 will be evaluated using super-resolution optical imaging techniques. Super-resolution imaging can be used to quantify the degree of CD20 clustering. Super-resolution imaging of biomaterial systems will more clearly link biological mechanisms to the overall effect induced by the biomaterial. The relationship between the crosslinking of CD20 and induction of apoptosis will be evaluated. Furthermore, the mechanism of action of a biomaterial can be determined by identifying other proteins or cellular structures involved. Optimized conjugates will then be tested in vivo and the results will be correlated with CD20 clustering results. This new therapeutic system will improve response rates in patients with NHL, and also may be applied to other B-cell related diseases such as multiple sclerosis and rheumatoid arthritis. The biorecognition of coiled-coil peptides at the cell surface to induce cellular signaling is an exciting strategy in treating diseases. Furthermore, using super-resolution imaging to evaluate self-assembly and protein crosslinking is a novel approach in evaluating therapeutics.

描述（由申请人提供）：该提案的目标是开发一种新的治疗系统以改善非霍奇金淋巴瘤（NHL）的治疗。自 1980 年以来，NHL 病例增加了一倍，2013 年估计有 69,000 例新诊断病例。B 细胞淋巴瘤占 NHL 病例的 85%；因此，新的抗癌疗法依赖于B细胞表面CD20受体的交联来诱导细胞凋亡。该治疗系统已被证明可以在体外和体内诱导 B 细胞凋亡。为了实现该项目的目标，我们将优化NHL治疗的治疗体系。该治疗系统利用一对带相反电荷的肽，形成反向平行的卷曲线圈。卷曲螺旋肽 (CC1/CC2) 通过生物识别形成物理交联。生物识别导致 B 细胞上 CD20 受体交联。 CD20 特异性 Fab' 片段连接到其中一个卷曲螺旋肽 (Fab'-CC1)，并且互补肽的多个拷贝连接到 N-(2-羟丙基)甲基丙烯酰胺 (HPMA) 共聚物 (P-CC2) ; P 是聚合物主链）。我们假设通过我们之前设计的缀合物结构修饰来增加 CD20 的交联将导致细胞凋亡增加。为了检验我们的假设并实现我们的目标，我们建议设计和合成不同结构的缀合物和含有 L-和/或 D-氨基酸残基的卷曲螺旋肽。这些缀合物将是高度可调的，并且由于肽之间的特定相互作用可以自组装成高度有序的结构。将评估缀合物生物识别并确定增强生物识别的设计参数。需要优化的结构特征包括：HPMA 主链的 Mw、肽接枝物的间距以及肽和聚合物/Fab' 之间的间隔长度。这些研究将为特定设计特征提供有价值的见解，从而改善一般盘绕线圈系统的自组装。将使用超分辨率光学成像技术评估细胞表面缀合物的自组装和 CD20 的交联。超分辨率成像可用于量化 CD20 聚类程度。生物材料系统的超分辨率成像将更清楚地将生物机制与生物材料引起的整体效应联系起来。将评估CD20的交联与细胞凋亡诱导之间的关系。此外，生物材料的作用机制可以通过识别涉及的其他蛋白质或细胞结构来确定。然后将在体内测试优化的缀合物，并将结果与 CD20 聚类结果相关联。这种新的治疗系统将提高 NHL 患者的缓解率，也可能应用于其他 B 细胞相关疾病，如多发性硬化症和类风湿性关节炎。在细胞表面对卷曲螺旋肽进行生物识别以诱导细胞信号传导是治疗疾病的一种令人兴奋的策略。此外，使用超分辨率成像来评估自组装和蛋白质交联是评估治疗的一种新方法。