Effect of RanbpM on amyloid pathology in a mouse model of Alzheimer's disease

RanbpM 对阿尔茨海默病小鼠模型淀粉样蛋白病理学的影响

基本信息

批准号：
8067642
负责人：
Madepalli Krishnappa Lakshmana
金额：
$ 3.22万
依托单位：
TORREY PINES INST FOR MOLECULAR STUDIES
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-01 至 2010-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8067642
关键词：
55-kDa Ran-binding protein Adaptor Signaling Protein Address Affect Age-Months Alzheimer&apos s Disease Americas Amino Acids Amyloid Amyloid Proteins Amyloid beta-Protein Amyloid beta-Protein Precursor Amyloid deposition Animal Model Binding Proteins Biochemical Biology Brain C-terminal Cell Culture Techniques Cells Cleaved cell Co-Immunoprecipitations Cytoplasmic Tail Data Deposition Disease Enzyme-Linked Immunosorbent Assay Foundations Future Grant Hippocampus (Brain)Hybrids Immunoblotting Intervention Intracellular Membranes Investigation LDL-Receptor Related Protein 1 Laboratories Late Onset Alzheimer Disease Length Ligand Binding Domain Ligands Link Lipoprotein Receptor Macroglobulins Mammalian Cell Membrane Membrane Protein Traffic Metabolism Mind Molecular Weight Mus Mutant Strains Mice N-terminal Nerve Degeneration Neurofibrillary Tangles Neurons Pathogenesis Pathology Patients Peptides Production Protein Binding Proteins Research Research Personnel Role SNAPIN gene Screening Result Senile Plaques Signal Transduction Site Testing Transfection Transgenic Mice Transgenic Organisms Variant Yeasts amyloid precursor protein processing apolipoprotein E-2 design extracellular in vivo insight mouse model multiple myeloma M Protein mutant novel novel therapeutics postnatal protein complex protein function protein metabolism protein transport public health relevance research study secretase stable cell line therapeutic target trafficking

项目摘要

DESCRIPTION (provided by applicant): The pathological hallmarks of Alzheimer's disease (AD) are the presence of senile plaques, largely composed of extracellular deposits of amyloid beta (A2) peptide and intracellular neurofibrillary tangles. Increasing evidence suggests that the low density lipoprotein receptor-related protein (LRP) facilitates amyloidogenic processing of APP. Within LRP, we defined the last 37 C-terminal residues of LRP (LRP-C37) lacking the NPxY or YxxL domains sufficient to robustly promote A2 production. The role of LRP-C37 domain in LRP function is largely unknown. Since LRP-C37 alone was a potent inducer of A2 production, we used this domain as bait in a yeast 2-hybrid screen, resulting in the identification of Ran-binding protein M (RanbpM), implicated in intracellular membrane trafficking and signaling. RanbpM interacted with both LRP and APP in transfected cells and mouse brain homogenates. Over-expression of RanbpM also enhanced the amount of LRP/APP complex formation in mammalian cells, suggesting a role for RanbpM as an adaptor protein that facilitates the tripartite interaction among APP, RanbpM and LRP. When 90 kDa full-length RanbpM (RanbpM-FL) was over-expressed in stable cell lines, a proteolytically cleaved N-terminal 55 kDa fragment (N-55) was also generated, which interacted strongly with both APP and LRP even more than the full-length RanbpM. This proteolytically cleaved species of RanbpM was elevated by 6-fold in brains of AD patients. Indeed, over-expression of RanbpM-FL or the N55 fragment markedly increased A2 secretion. We hypothesize that RanbpM alters APP metabolism in vivo, in particular via the N55 fragment of RanbpM. The specific aims of this application are to first generate transgenic mice with neuronal expression of RanbpM-FL and RanbpM-N55 and then to cross with mice over-expressing APP to obtain RanbpM/APP double transgenic mice. Whether over-expression of RanbpM variants alter amyloidogenic processing of APP will be determined by quantifying the levels of A2 and amyloid plaques by biochemical and immunohistochemical examinations in the cortex and hippocampus of Tg RanbpM/APP double transgenic mice. The level of CHAPS-soluble and insoluble A2 will be quantified by ELISA at 2 and 10 months of age. The characterization of novel APP and LRP-interacting proteins that promote amyloidogenic processing of APP will reveal novel sites of pathogenesis and targets for anti-amyloid intervention. We believe that RanbpM represents a potential therapeutic target. In addition, the RanbpM transgenic mice generated through this R03 granting mechanism is expected to hold substantial extrinsic merit, as they will be undoubtedly be useful to many researchers who study other aspects of RanbpM. This application may also lay the foundation for a future R01 investigation on the role of RanbpM in neurodegeneration. PUBLIC HEALTH RELEVANCE: Alzheimer's disease (AD), a disease characterized by toxic amyloid (A2) accumulation in brain, affects about 5 million people in America, but despite extensive research, available treatments provide only limited symptomatic relief. Recently we found a protein called RanbpM that enhances the production of amyloid/A2 in cells cultured in the laboratory. In this application, we will express RanbpM in brains of mice to confirm the findings in animal models in the hopes of that new insights may be uncovered for therapeutically targeting amyloid production in AD, in particular with RanbpM as a therapeutic target in mind.

描述（由申请人提供）：阿尔茨海默氏病（AD）的病理标志是存在老年斑块，这很大程度上由淀粉样蛋白β（A2）肽和细胞内神经原纤维缠结的细胞外沉积物组成。越来越多的证据表明，低密度脂蛋白受体相关蛋白（LRP）促进了APP的淀粉样生成加工。在LRP中，我们定义了缺乏NPXY或YXXL结构域的LRP（LRP-C37）的最后37个C末端残基，足以稳健地促进A2产生。 LRP-C37域在LRP功能中的作用在很大程度上未知。由于仅LRP-C37是A2产生的有效诱导剂，因此我们将该结构域用作酵母2杂化筛网中的诱饵，从而鉴定了RAN结合蛋白M（RANBPM），与细胞内膜的运输和信号传导有关。 RANBPM在转染的细胞和小鼠脑匀浆中与LRP和APP相互作用。 RANBPM的过表达还增强了哺乳动物细胞中LRP/APP复合物形成的量，这表明RANBPM作为一种衔接蛋白的作用，可促进APP，RANBPM和LRP之间的三方相互作用。当90 kDa全长ranbpm（ranbpm-fl）在稳定的细胞系中过表达时，还会产生一种蛋白水解切割的N末端55 kDa片段（N-55），与App和LRP相互作用甚至超过App和LRP全长ranbpm。在AD患者的大脑中，这种蛋白水解的RANBPM裂解物种升高了6倍。实际上，RANBPM-FL或N55片段的过表达显着增加了A2分泌。我们假设RANBPM在体内尤其是通过RANBPM的N55片段来改变APP代谢。该应用程序的具体目的是首先生成带有RANBPM-FL和RANBPM-N55的神经元表达的转基因小鼠，然后与过度表达的应用程序交叉以获得RANBPM/APP双转基因小鼠。 RANBPM变体的过度表达是否会改变APP的淀粉样生成处理，这将通过在TG RANBPM/APP APP/App Double Transgenic小鼠的皮质和海马中量化A2和淀粉样蛋白斑的水平。 ELISA将在2个月和10个月大时量化CHAPS可溶性和不溶性A2的水平。促进APP的淀粉样生成加工的新型APP和LRP相互作用蛋白的表征将揭示发病机理的新颖部位和抗淀粉样蛋白干预的靶标。我们认为RANBPM代表了潜在的治疗靶点。此外，通过这种R03授予机制产生的RANBPM转基因小鼠有望具有巨大的外部功绩，因为它们无疑对许多研究RANBPM其他方面的研究人员有用。该应用程序还可能为R01对RANBPM在神经变性中的作用进行研究奠定基础。公共卫生相关性：阿尔茨海默氏病（AD）是一种以毒性淀粉样蛋白（A2）积累为特征的疾病，影响了美国约500万人，但是尽管进行了广泛的研究，但可用的治疗方法仅提供有限的症状缓解。最近，我们发现了一种称为RANBPM的蛋白质，可增强实验室培养的细胞中淀粉样蛋白/A2的产生。在此应用中，我们将在小鼠的大脑中表达RANBPM，以确认动物模型中的发现，以期希望可以发现新见解以治疗靶向AD的淀粉样蛋白产生，尤其是用RANBPM作为治疗靶标的。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

RanBP9 Plays a Critical Role in Neonatal Brain Development in Mice.

RanBP9 在小鼠新生大脑发育中发挥着关键作用。

DOI：
10.1371/journal.pone.0066908
发表时间：
2013
期刊：
PloS one
影响因子：
3.7
作者：
Palavicini,JuanPablo;Lloyd,BrandonNoel;Hayes,CrystalD;Bianchi,Elisabetta;Kang,DavidE;Dawson-Scully,Ken;Lakshmana,MadepalliK
通讯作者：
Lakshmana,MadepalliK

RanBP9 overexpression down-regulates phospho-cofilin, causes early synaptic deficits and impaired learning, and accelerates accumulation of amyloid plaques in the mouse brain.

RanBP9 过度表达会下调磷酸丝切蛋白，导致早期突触缺陷和学习受损，并加速小鼠大脑中淀粉样斑块的积累。

DOI：
10.3233/jad-131550
发表时间：
2014
期刊：
Journal of Alzheimer's disease : JAD
影响因子：
0
作者：
Palavicini,JuanPablo;Wang,Hongjie;Minond,Dmitriy;Bianchi,Elisabetta;Xu,Shaohua;Lakshmana,MadepalliK
通讯作者：
Lakshmana,MadepalliK

RanBP9 overexpression accelerates loss of dendritic spines in a mouse model of Alzheimer's disease.

DOI：
10.1016/j.nbd.2014.05.029
发表时间：
2014-09
期刊：
Neurobiology of disease
影响因子：
6.1
作者：
Wang R;Palavicini JP;Wang H;Maiti P;Bianchi E;Xu S;Lloyd BN;Dawson-Scully K;Kang DE;Lakshmana MK
通讯作者：
Lakshmana MK

RanBP9 overexpression reduces dendritic arbor and spine density.