Conformational properties of misfolded protein aggregates in cases involving the co-occurrence of prion disease and Alzehimer's disease or prion disease and CTE

项目摘要/摘要 多种致病蛋白同时出现在获得性和特发性疾病中是一种常见的事件。 神经退行性疾病。淀粉样β蛋白(A-β)和tau蛋白在阿尔茨海默病中共存；α-突触核蛋白 在帕金森氏病痴呆患者中为β；在皮质基底部变性患者中为TARDNA结合蛋白43和tau。 最近，我研究了医源性CreutzFeldt-Jakob病(ICJD)，这是一种通过感染获得的人类Pron病， 慢性创伤性脑病/创伤后应激障碍(CTE/PTSD)，一种神经退行性疾病 继发于反复创伤的情况。ICJD的一个子集的特点是致病或疾病共同出现- 相关的普恩蛋白(Prpd)和Aβ，我认为它们都是人传人的结果。 同样，CTE/PTSD的一个子集含有PRPD和致病性tau(TauD)。多个因素并存 神经退行性变条件下的致病蛋白尚未完全了解。 在这篇K99/R00中，我建议研究iCJD共病蛋白病的分子特征。 散发性CJD(SCJD)和CTE/PTSD以及动物模型。医源性CJD和CTE/PTSD提供 在研究非年龄相关多蛋白神经退行性变机制方面具有独特优势。三 提出了相关的具体目标。具体目标1重点研究PrPD和Aβ的构象性质 从与β病理相关的iCJD和sCJD病例与β中的PRPD特征进行比较- 晚发性阿尔茨海默病将提供经典Aβ的对照数据。特定目标 2从患有这两种蛋白病变的CTE/PTSD病例中获得的PRPD和tauD的构象特征。 与受tauD而不是PRPD影响的相应的CTE/PTSD病例相比，Pron病的病例将 作为经典的PRPD控制。为这些研究提出的方法学包括基于质谱学的 评估共存致病蛋白构象特征的方法，播种动力学 蛋白质错折叠循环扩增(PMCA)技术及原代神经元细胞毒性检测 文化。特指目标3进一步剖析了通过传递 新型共表达人细胞PrP和人PrP转基因小鼠模型的上述条件 淀粉样前体蛋白(APP)。生物测定将检查多蛋白的关键动态方面。 神经退行性变，包括传播的时间和途径，是否存在病原菌的相互作用 繁殖过程中的蛋白质和疾病表型形成的阶段。我相信这些加在一起 利用新描述的包含多蛋白和单蛋白病变的条件的研究 通过新的实验模型，将在一个重要的领域产生重要和所需的信息 研究。通过这些研究，在我的两位主要导师Surewicz博士的悉心指导下 朱，以及我的指导委员会的其他成员，我将获得所需的经验和技能 成为一名成功的独立研究者和神经退行性疾病领域的领导者。

PROJECT SUMMARY/ABSTRACT The co-occurrence of more than one pathogenic protein is a frequent event in acquired and idiopathic neurodegenerative diseases. Amyloid beta (Aβ) and tau pathologies coexist in Alzheimer's disease; α-synuclein and Aβ in Parkinson's disease with dementia; TAR DNA-binding protein 43 and tau in corticobasal degeneration. Recently, I studied iatrogenic Creutzfeldt-Jakob disease (iCJD), a human prion disease acquired by infection, and chronic traumatic encephalopathy/post-traumatic stress disorder (CTE/PTSD), a neurodegenerative condition secondary to repetitive trauma. A subset of iCJD featured the co-occurrence of pathogenic or disease- related prion protein (PrPD) and Aβ, both of which I suggested result from human-to-human transmission. Similarly, a subset of CTE/PTSD harbored PrPD and pathogenic tau (tauD). The co-existence of multiple pathogenic proteins in neurodegenerative conditions has not yet been fully understood. In this K99/R00 I propose to investigate molecular characteristics of co-occurring proteinopathies in iCJD, sporadic CJD (sCJD) and CTE/PTSD as well as in animal models. Iatrogenic CJD and CTE/PTSD offer the unique advantage to study the mechanisms of non-age related multiprotein neurodegeneration. Three interrelated specific aims are proposed. Specific aim 1 focuses on conformational properties of PrPD and Aβ aggregates from cases with iCJD and sCJD associated with Aβ pathology compared to PrPD features in Aβ- negative iCJD and sCJD; late onset Alzheimer's disease will provide control data from classic Aβ. Specific aim 2 deals with conformational features of PrPD and tauD harvested from CTE/PTSD cases with both proteinopathies compared to corresponding CTE/PTSD cases affected by tauD but not PrPD; cases of prion disease will serve as classic PrPD controls. The methodologies proposed for these studies comprise mass spectrometry-based approaches to assess the conformational features of the co-existing pathogenic proteins, seeding kinetics by protein misfolding cyclic amplification (PMCA) technology and cytotoxicity assay employing primary neuronal cultures. Specific aim 3 dissects further aspects of the co-occurring proteinopathies by transmission of the aforementioned conditions to novel transgenic mouse models co-expressing the human cellular PrP and human amyloid precursor protein (APP). The bioassay will examine critical dynamic aspects of multiprotein neurodegeneration, including timing and route of propagation, existence of interactions of the pathogenic proteins during propagation and stages in the formation of the disease phenotype. I believe that together these studies that take advantage of newly-described conditions comprising multi- and single- proteinopathies along with novel experimental models, will generate significant and needed information on an important area of research. Through these studies and under the dedicated guidance of my two primary mentors, Drs. Surewicz and Zhu, and of the other members of my mentoring committee, I will acquire the experience and skills needed to become a successful independent investigator and a leader in the field of neurodegeneration.