Elucidating the cellular mechanisms of prion propagation and clearance for devisi

描述(申请人提供)：阐明Pron增殖和清除的细胞机制以设计干预Pron疾病的新靶点越来越多的神经退行性疾病是由错误折叠的蛋白质聚集引起的，它们具有共同的病理生理机制。Prion病是典型的蛋白质错误折叠疾病，其发病机制仅与单个细胞蛋白(PrPc)的异常错误折叠有关。Prion病在这组中是独一无二的，因为 它们是在人和动物身上发现的感染性疾病。除了零星或遗传表现外，它们还可通过感染获得，并在物种之间传播，导致地方性或流行性情况(例如疯牛病/vCJD和CWD)。它们是可以控制的，但根除是不可能的。因此，为了设计合理的策略来控制这些事件，必须了解病毒传播和传播的分子和细胞需求。了解蛋白错误折叠的病理生理学方面的进展将对其他蛋白质错误折叠疾病具有重要意义，因为它可能有助于阐明常见的细胞机制。这种理解具有根本性的科学意义，因为神经退行性疾病是我们老龄化社会中最大的健康问题之一，而揭示普遍有效性的分子机制对于识别新的靶点和开发合理的治疗方法是至关重要的。我们小组的长期目标是开发治疗性和预防性的反普鲁恩策略。我们的总体目标是研究Pron感染的细胞和分子生物学，并利用所获得的了解来描述新的干预目标。我们将我们的尝试集中在两个主要战略上。一种是细胞内源性的PrPc清除能力，另一种是靶向细胞内PrPc的异构体，PrPc是PrPc转化和神经退行性变的先决条件。我们的中心假设是，通过增加细胞数量来干扰病毒的传播是可行的。 普里恩病毒的许可。目标1的工作将证实我们的发现，即通过化合物诱导的自噬--一种降解和循环的基本细胞程序--可以增强普恩清除。这项拟议的工作旨在更好地了解潜在的分子机制，并验证这一发现在体内的治疗和翻译潜力。目标2和目标3中的工作解决了普恩形成的细胞修饰物。我们发现基础水平的自噬是建立Pron感染所必需的，我们认为自噬在生物学上等同于哺乳动物类Pron/Prion生物学中假定的解聚酶功能。我们的目标是在细胞和分子水平上证明这一点。目标3的工作理由是，分泌途径中的蛋白质质量控制机制可以通过决定转化底物的质量来直接影响普恩转化。我们想通过过度表达折叠和排序蛋白质来操纵这一点，以表明这代表了一种新的对抗Pron传播的途径。总体而言，我们的研究将提供对与神经退行性疾病相关的基本细胞和分子机制的机械性见解，并将导致合理治疗Pron疾病和蛋白质错误折叠障碍的新靶点。

DESCRIPTION (provided by applicant): Elucidating the cellular mechanisms of prion propagation and clearance for devising new targets for intervention in prion disease There are an increasing number of neurodegenerative disorders which result from the aggregation of misfolded proteins and which share patho-physiological mechanisms. Prion diseases are the prototypical protein misfolding diseases, and their pathogenesis is associated solely with aberrant misfolding of a single cellular protein (PrPc). Prion diseases are unique in this group as they are infectious disorders found in man and animals. Besides sporadic or genetic manifestation, they can be acquired by infection and transmitted between species, resulting in endemic or epidemic scenarios (e.g. BSE/vCJD and CWD). They can be controlled, but eradication is impossible. Therefore, it is mandatory to understand the molecular and cellular requirements for propagation and transmission of prions in order to device rational strategies for controlling these events. Advances in understanding prion patho-physiology will have major implications for other protein misfolding diseases, as it may help elucidate common cellular mechanisms. Such understanding is of fundamental scientific importance as neurodegenerative diseases represent one of the biggest health problems in our aging society, and uncovering molecular mechanisms of general validity is fundamental for the identification of new targets and development of rational therapies. The long-term goal of our group is to develop therapeutic and prophylactic anti-prion strategies. The overall objective we have is to study the cellular and molecular biology of prion infections and to use gained understanding for delineating novel targets for intervention. We have focused our attempts on two main strategies. One is the endogenous cellular clearance capacity for prions, the other one is to target the cellular isoform PrPc, which is a prerequisite for prion conversion and execution of neurodegeneration. It is our central hypothesis that it is feasible to interfere in prion propagation by increasing the cellular clearance for prions. Work in Aim 1 will substantiate our finding that prion clearance can be enhanced by compound-induced induction of autophagy, a basic cellular program for degradation and recycling. The proposed work intends to better understand the underlying molecular mechanisms and to validate the therapeutic and translational potential of this finding in vivo. Work in Aim 2 and 3 addresses cellular modifiers of prion formation. We have found that a basal level of autophagy is needed for establishing prion infection and we propose that autophagy represents the biological equivalent for the postulated disaggregase function in mammalian prion/prion-like biology. Our goal is to prove this at the cellular and molecular level. The rational for work in Aim 3 is that protein quality control mechanisms in the secretory pathway can directly influence prion conversion by determining on the quality of conversion substrates. We want to manipulate this by over-expressing folding and sorting proteins, in order to show that this represents a novel pathway counteracting prion propagation. Overall, our studies will provide mechanistic insights into basic cellular and molecular mechanisms which are relevant for neurodegenerative diseases and will result in novel targets for rational therapy against prion diseases and protein misfolding disorders.