Targeted Delivery of S100B Inhibitory Peptide to SCA1 Mouse Cerebellum

S100B 抑制肽靶向递送至 SCA1 小鼠小脑

• 批准号: 7990138
• 负责人: DRAZEN RAUCHER
• 金额: $ 7.45万
• 依托单位: UNIVERSITY OF MISSISSIPPI MED CTR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7990138
• 关键词: Address; Amino Acid Sequence; Ataxia; Binding; Biopolymers; Blood - brain barrier anatomy; Brain Stem; Brain region; CAG repeat; Cell Nucleus; Cells; Cellular Morphology; Central Nervous System Diseases; Cerebellar Ataxia; Cerebellum; Cessation of life; Clinical Trials; Dendritic Spines; Development; Drug Kinetics; Elastin; Engineering; Fever; Figs - dietary; Genes; Goals; Heating; Impairment; Intravenous; Kinetics; Label; Local Hyperthermia; Measures; Mediating; Modeling; Motor; Mus; Neurodegenerative Disorders; Neurophysiology - biologic function; Neuroprotective Agents; Pathology; Peptides; Phosphorylation; Physiological; Plasma; Proteins; Purkinje Cells; Radio; Research; Route; Signal Transduction; Site; Solutions; Stress; TRTK-12 peptide; Technology; Temperature; Therapeutic; Tissues; Toxic effect; Treatment Efficacy; Type 1 Spinocerebellar Ataxia; Vacuole; Work; aqueous; ataxin-1; bactenecin; base; cell type; design; effective therapy; improved; in vivo; interest; mouse model; mutant; nervous system disorder; new technology; novel; polyglutamine; polypeptide; pre-clinical; prevent; public health relevance; response; targeted delivery; tool

DESCRIPTION (provided by applicant): Spinocerebellar ataxia-1 (SCA1) is a devastating neurological disorder, resulting from CAG repeat expansion in the ataxin-1 gene. The polyglutamine expanded mutant ataxin-1 primarily targets Purkinje cells (PCs) of the cerebellum. The exact mechanism of PC degeneration in SCA1 is poorly understood, but it is known that the earliest morphologic change seen in SCA1 PCs is the development of cytoplasmic vacuoles that contain Bergmann glial (BG) proteins, especially S100B. These vacuoles are toxic and alter PC morphology, and may be forming in response to a stress signal of PCs. Furthermore, S100B released by BG or from the vacuoles may modulate Akt phosphorylation of mutant ataxin-1. Akt is known to stabilize mutant ataxin-1 to aggregate in the nucleus of PCs. The objective of the proposed study is to target SCA1 PCs with therapeutic peptide (TP) that will influence ataxin-1 aggregation and toxicity. Though TPs are designed to inhibit specific molecular interactions, their efficacy in vivo is limited by poor pharmacokinetic parameters. To improve their pharmacokinetics and bio-distribution, TP will be fused to a thermally responsive polypeptide-based carrier. This polypeptide can be targeted to the cerebellum of Tg mice by applying local hyperthermia. The amino acid sequence of the thermally responsive polypeptide is based on elastin-like polypeptide (ELP) biopolymers, which are soluble in aqueous solution below physiological temperature, but aggregate when the temperature is raised above 39oC. A cell-penetrating peptide (CPP) is conjugated to the ELP to enhance delivery of the polypeptide across the blood brain barrier (BBB) and to facilitate cell entry. To the CPP-ELP, therapeutic peptide TRTK-12, which interacts with S100B will be conjugated. This TP will be administered intranasally (IN) or intraperitoneally (IP) and, by applying local hyperthermia, it will be demonstrated that these genetically engineered polypeptides can be targeted to a diseased site and improve motor coordination and cerebellar pathology in a SCA1 mouse model. This project will address the following specific aims: Specific Aim 1: Determine feasibility of delivering Bac-ELP or SynB1-ELP IN or IP to the cerebellum. The plasma kinetics and the in vivo distribution of radio-labeled Bac/SynB1-ELP will be measured in the cerebellum, other brain regions and tissues in a SCA1 mouse with and without hyperthermia. Specific Aim 2: Evaluate the therapeutic efficacy TRTK-12 attached to the optimal CPP-ELP as determined in Specific Aim 1. It is anticipated that TP treatment will result in improved motor coordination and PC pathology and will provide preclinical support for the therapeutic potential of this novel technology. Though the SCA1 mouse will be used as a model in the proposed work, this approach could also be applied to thermal targeting of ELP-fused TPs to many other CNS disorders. Therefore, successful completion of the proposed research may have a significant impact, by not only propelling this technology into clinical trials, but also providing a powerful tool to treat and manage other CNS diseases. PUBLIC HEALTH RELEVANCE: Spinocerebellar ataxia-1 (SCA1) is a devastating neurological disease resulting from CAG repeat expansion in the ataxin-1 gene, leading to polyglutamine expansion at the protein level, and degeneration of Purkinje cells as well as some other cell types in the brainstem. Currently, there is no specific treatment to delay or halt the progression of SCA1. We plan to use a very unique strategy of thermally targeted delivery of candidate therapeutic peptides to the cerebellum via intranasal or intravenous routes with the goal of developing useful new strategies for treating SCA1 and other ataxias.

描述（由申请人提供）：脊髓脑性共济失调-1（SCA1）是一种毁灭性的神经系统疾病，是由acaxin-1基因中的CAG重复扩张引起的。聚谷氨酰胺扩展了突变体共济素1主要靶向小脑的Purkinje细胞（PC）。 SCA1中PC变性的确切机制知之甚少，但众所周知，SCA1 PC中最早的形态变化是含有伯格曼神经胶质（BG）蛋白（尤其是S100B）的细胞质液泡的发展。这些液泡有毒并改变了PC的形态，可能会响应PC的应力信号而形成。此外，由BG或从液泡中释放的S100B可能会调节突变蛋白1的Akt磷酸化。已知AKT可以稳定突变体ataxin-1以在PC的核中聚集。拟议的研究的目的是针对具有治疗肽（TP）的SCA1 PC，这将影响ataxin-1的聚集和毒性。尽管TPS旨在抑制特定的分子相互作用，但它们在体内的功效受到药代动力学参数不良的限制。为了改善其药代动力学和生物分布，TP将融合到热响应的基于多肽的载体上。该多肽可以通过施加局部高温靶向TG小鼠小脑。热响应性多肽的氨基酸序列基于弹性蛋白样多肽（ELP）生物聚合物，它们可在生理温度以下的水溶液中溶于水溶液中，但是当温度升高39oC时，聚集了。将细胞穿透肽（CPP）偶联到ELP中，以增强多肽在血液脑屏障（BBB）上的递送并促进细胞进入。对于CPP-ELP，将结合与S100B相互作用的治疗肽TRTK-12。该TP将用于鼻内（IP）或腹膜内（IP），并且通过应用局部高温，可以证明这些基因工程性多肽可以靶向患病部位并改善运动率和小脑病理学在SCA1小鼠模型中。该项目将解决以下特定目的：具体目标1：确定将BAC-ELP或SYNB1-ELP或IP输送到小脑的可行性。在具有和不含超热的SCA1小鼠中，将在小脑，其他大脑区域和组织中测量无线电标记的BAC/SYNB1-ELP的血浆动力学和体内分布。特定目的2：评估特定目标中确定的最佳CPP-ELP的治疗功效TRTK-12。预计TP治疗将改善运动配位和PC病理学，并为这项新技术的治疗潜力提供临床前的支持。尽管SCA1小鼠将在拟议的工作中用作模型，但该方法也可以应用于将ELP融合的TPS的热靶向到许多其他CNS疾病中。因此，拟议的研究的成功完成可能会产生重大影响，不仅将该技术推向临床试验，而且还提供了一种强大的工具来治疗和管理其他CNS疾病。 公共卫生相关性：脊髓脑性共济失调-1（SCA1）是一种毁灭性的神经系统疾病，是由于CAG重复膨胀而导致蛋白质水平上的多谷氨酰胺扩展，以及Purkinje细胞的多谷氨酰胺扩展以及大脑中的其他细胞类型。当前，没有特定的治疗方法可以延迟或停止SCA1的进展。我们计划使用非常独特的策略，即通过鼻内或静脉内路线将候选治疗肽的热靶向输送到小脑，目的是制定有用的新策略来治疗SCA1和其他共济失调。