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: 8071629
• 负责人: DRAZEN RAUCHER
• 金额: $ 7.32万
• 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 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8071629
• 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; 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; Toxic effect; Treatment Efficacy; Type 1 Spinocerebellar Ataxia; Vacuole; Work; aqueous; ataxin-1; bactenecin; base; brain tissue; 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）：脊髓小脑共济失调-1 （SCA1）是一种毁灭性的神经系统疾病，由共济失调素-1基因CAG重复扩增引起。聚谷氨酰胺扩增突变体ataxin-1主要靶向小脑浦肯野细胞（PCs）。SCA1中PC变性的确切机制尚不清楚，但已知SCA1中PC最早的形态学改变是细胞质空泡的发育，其中含有伯格曼胶质（BG）蛋白，特别是S100B。这些液泡是有毒的，并改变PC形态，可能是在PC的应激信号下形成的。此外，BG或液泡释放的S100B可能调节突变体ataxin-1的Akt磷酸化。已知Akt稳定突变体ataxin-1在PCs细胞核中聚集。该研究的目的是用治疗肽（TP）靶向SCA1 PCs，这将影响ataxin-1的聚集和毒性。虽然TPs被设计为抑制特定的分子相互作用，但它们在体内的功效受到不良药代动力学参数的限制。为了改善它们的药代动力学和生物分布，TP将被融合成一种热响应的多肽载体。该多肽可通过局部热疗作用于Tg小鼠小脑。热响应多肽的氨基酸序列以弹性蛋白样多肽（ELP）生物聚合物为基础，在生理温度以下可溶于水溶液，但在温度高于39℃时聚集。细胞穿透肽（CPP）与ELP结合，以增强多肽穿过血脑屏障（BBB）的传递，并促进细胞进入。将与S100B相互作用的治疗肽TRTK-12偶联到CPP-ELP上。这种TP将通过鼻内（IN）或腹腔内（IP）给药，通过局部热疗，将证明这些基因工程多肽可以靶向病变部位，改善SCA1小鼠模型的运动协调和小脑病理。该项目将解决以下具体目标：具体目标1：确定将Bac-ELP或SynB1-ELP IN或IP输送到小脑的可行性。将在有和没有高温的SCA1小鼠的小脑、其他脑区和组织中测量放射性标记Bac/SynB1-ELP的血浆动力学和体内分布。特异性目的2：评估特异性目的1确定的最佳pcp - elp附着TRTK-12的治疗效果。预计TP治疗将改善运动协调和PC病理，并将为这种新技术的治疗潜力提供临床前支持。虽然SCA1小鼠将在拟议的工作中用作模型，但这种方法也可以应用于elp融合的tp的热靶向治疗许多其他中枢神经系统疾病。因此，该研究的成功完成可能会产生重大影响，不仅推动该技术进入临床试验，而且还为治疗和管理其他中枢神经系统疾病提供了强有力的工具。

项目成果

Glial S100B protein modulates mutant ataxin-1 aggregation and toxicity: TRTK12 peptide, a potential candidate for SCA1 therapy.

• DOI: 10.1007/s12311-011-0262-5
• 发表时间: 2011-06
• 期刊: CEREBELLUM
• 影响因子: 3.5
• 作者: Vig, Parminder J. S.;Hearst, Scoty;Shao, Qingmei;Lopez, Mariper E.;Murphy, Henry A., II;Safaya, Eshan
• 通讯作者: Safaya, Eshan