Targeting Rab6-km23-1-mediated compartmentalized trafficking as a novel therapeutic approach to Alzheimers Disease

靶向 Rab6-km23-1 介导的区室化运输作为阿尔茨海默病的新型治疗方法

• 批准号: 9756263
• 负责人: Lance Allen Liotta
• 金额: $ 19.14万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9756263
• 关键词: Age; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid; Amyloid beta-Protein Precursor; Animal Model; Area; Binding; Brain; C-terminal; Carrier Proteins; Cell model; Cells; Clinical Trials; Complex; Computer Simulation; DNA Sequence Alteration; Dendrites; Down Syndrome; Drug Targeting; Dynein ATPase; Endosomes; Engineering; Enzymes; Event; Failure; Family; Functional disorder; Golgi Apparatus; Growth; Hippocampus (Brain); Hot Spot; Human; Individual; Lead; Light; Location; MAP Kinase Gene; Mediating; Mediator of activation protein; Modeling; Motor; Mus; Mutation; Nerve; Neurites; Neurons; Organelles; Outcome; Paint; Pathogenesis; Pathology; Pathway interactions; Patients; Peptides; Play; Presenile Alzheimer Dementia; Prevention strategy; Process; Production; Protein Family; Protein Fragment; Protein Region; Proteins; Proteolysis; Recycling; Regulation; Rodent; Role; Route; Senile Plaques; Signal Transduction; Site; Sorting - Cell Movement; Structural Models; Surface; Synapses; Technology; Testing; Therapeutic; Therapeutic Agents; Transport Vesicles; Vesicle; Vesicle Transport Pathway; alpha secretase; amyloid peptide; amyloid precursor protein processing; axonopathy; base; beta secretase; beta-site APP cleaving enzyme 1; design; disease phenotype; familial Alzheimer disease; human model; induced pluripotent stem cell; inhibitor/antagonist; link protein; member; mutant; neuron loss; neuronal cell body; next generation; novel; novel strategies; novel therapeutic intervention; novel therapeutics; peptide drug; prevent; protein complex; protein protein interaction; protein transport; rab GTP-Binding Proteins; retrograde transport; spatiotemporal; synthetic peptide; targeted treatment; therapeutic target; trafficking

Summary The amyloid hypothesis as the cause for Alzheimer's Disease (AD) has recently come under fire due to the failure of so many clinical trials for amyloid peptide (Aß)-targeting therapies. However, it may be that amyloid precursor protein (APP) itself, or the C-terminal fragment-ß (CTFß) produced by ß-secretase (BACE1) cleavage, is the actual culprit in AD, having a more direct role in AD than previously thought. In the current proposal, we will study a novel approach to reducing early-stage AD by targeting the protein-protein interaction of a complex (km23-1–Rab6) that we propose controls the trafficking of APP and the rate-limiting enzyme (BACE1) in APP processing. More specifically, we will test the novel hypothesis that APP and BACE1 are transported in km23-1–Rab6 vesicles to the Golgi, where CTFß is produced to play a role in neurite outgrowth and dendritic branching. We hypothesize, further, that APP or CTFß accumulation here in AD causes exuberant and aberrant hippocampal axodentritic sprouting, eventually leading to diminish dendritic arbor complexity and breakage of neuronal branches. We will examine the km23-1–Rab6-mediated transport of APP and BACE1 to the Golgi, as well as the effects of the resulting APP fragments on dendritic branching. The results will provide a stronger basis for targeting the physical association of APP and BACE1 at Golgi sites, to reduce BACE1 activity and the subsequent production of APP intermediates. Α greater understanding of the mechanisms underlying km23-1–Rab6 regulation of APP and BACE1 interactions at the Golgi to control dendritic arborization should facilitate targeting this key trafficking event to reduce early AD-associated causal events. Our structural modeling of the km23-1–Rab6 complex revealed “hot spots” for the precise sites of protein- protein interaction. In Aim 1, we will examine the effects of site-specific mutants of the proteins on the spatiotemporal regulation of km23-1–Rab6 complexes, as well as on APP/BACE1 association and compartment location. The focus will be on APP and BACE1 association at soma Golgi, as well as at Golgi outposts (GOs) in developing dendrites, in order to better understand the mechanisms underlying APP trafficking, processing, and signaling at these specific regions. In Aims 2 and 3, we will us in silico modeling and apply a novel Protein Painting technology to reveal the unique interface by which km23-1 interacts with Rab6 in regulating APP trafficking and processing. The precise interaction region will be used to design corresponding peptide inhibitors to be tested for inhibitory effects on km23-1–Rab6 complex formation, APP processing, and AD-associated pathologies. While the majority of previous studies have used rodent familial AD (FAD) models, here we will apply human models that recapitulate sporadic AD (sAD) to test our novel km23-1–Rab6 inhibitors. The use of the hidden contact regions between these critical interacting proteins as drug targets will lead to paradigm-shifting therapies, overcoming the limitations with past therapeutic strategies. The novel therapeutic agents for AD developed as a result of the proposed studies will be among critical members of the next generation of AD-targeted therapeutics.

总结 淀粉样蛋白假说作为阿尔茨海默病（AD）的原因，最近受到抨击，由于 许多针对淀粉样肽（AAPs）的临床试验失败。然而，淀粉样蛋白 前体蛋白（APP）本身，或β-分泌酶（BACE 1）产生的C-末端片段-β-内酰胺酶（CTF 3） 卵裂是AD的实际罪魁祸首，在AD中具有比以前认为的更直接的作用。在当前 我们将研究一种新的方法，通过靶向蛋白质-蛋白质相互作用来减少早期AD 我们提出的复合物（km 23 -1-Rab 6）控制APP和限速酶的运输 （BACE 1）在APP处理中。更具体地说，我们将测试APP和BACE 1是 在km 23 -1-Rab 6囊泡中转运到高尔基体，在高尔基体中产生CTF 3，在神经突生长中发挥作用 和树枝状分支。我们进一步假设，AD中APP或CTF β的积累导致 海马轴突出芽异常旺盛，最终导致树突乔木减少 神经分支的复杂性和断裂。我们将研究km 23 -1-Rab 6介导的APP转运 和BACE 1对高尔基体的作用，以及由此产生的APP片段对树突状分支的影响。的 结果将为APP和BACE 1在高尔基体位点的物理结合提供更有力的基础， 降低BACE 1活性和随后APP中间体的产生。更好地理解 km 23 -1-Rab 6调控高尔基体APP和BACE 1相互作用的机制， 树突状树枝化应有助于靶向这一关键的贩运事件，以减少早期AD相关的因果关系。 事件 我们对km 23 -1-Rab 6复合物的结构建模揭示了蛋白质精确位点的“热点”， 蛋白质相互作用在目标1中，我们将研究蛋白质的位点特异性突变体对蛋白质表达的影响。 km 23 -1-Rab 6复合物的时空调节，以及APP/BACE 1缔合， 隔间位置。重点将是APP和BACE 1协会在索马高尔基体，以及在高尔基体 前哨（英语：Outposts）在树突发育中的作用，以便更好地理解APP的机制 在这些特定区域进行贩运、加工和信号传递。在目标2和3中，我们将使用计算机建模 并应用一种新的蛋白质绘画技术来揭示km 23 -1与 Rab 6在规范APP贩运和处理方面的作用。精确的相互作用区域将用于设计 测试相应肽抑制剂对km 23 -1-Rab 6复合物形成的抑制作用，APP 加工和AD相关病理。虽然以前的大多数研究都使用啮齿动物家族 AD（FAD）模型，在这里我们将应用概括散发性AD（sAD）的人类模型来测试我们的新的 km 23 -1-Rab 6抑制剂。利用这些关键的相互作用蛋白质之间的隐藏接触区域， 药物靶点将导致范式转变疗法，克服过去治疗策略的局限性。 作为拟议研究的结果，开发的新型AD治疗药物将是关键的 下一代AD靶向治疗的成员。