The sex-specific role of alpha-synuclein

摘要 生物学变量可以直接影响疾病的脆弱性和临床表现。例如，年龄仍然 是发展大多数神经退行性疾病的最大风险因素，虽然性别差异 虽然在历史上没有得到充分的研究，但越来越多的证据表明，性也会影响疾病的结果。阿尔法 突触核蛋白（αSyn）长期以来与许多神经退行性疾病的发展有关， 然而，它的一些正常职能仍然难以捉摸。αSyn是突触核蛋白家族蛋白质的一部分，是一种高度保守的蛋白质。 进化上保守的一组，包括β（βSyn）和γ突触核蛋白。αSyn和βSyn是 越来越多地被研究为疾病发展和进展的潜在生物标志物。βSyn还 已被证明是αSyn聚集的内源性抑制剂，并且可以响应于 删除αSyn。最近的研究表明，突触核蛋白的蛋白质丰度和功能可能随性别和年龄而变化。 年龄然而，很少有研究严格评估这些生物变量在正常情况下的相互作用。 功能系统。如果不了解这些蛋白质的基线丰度和功能， 靶向突触核蛋白家族的药物在临床上失败或产生负面影响的可能性增加 方面的影响.我们小组的结果显示，小鼠对αSyn消融的反应具有性别特异性，因此雌性 动物而非雄性动物的空间学习和记忆受损。本提案的目的是调查 在多个年龄段，两性的αSyn功能正常。总的假设是αSyn是 男性和女性的大脑在转录上、功能上或功能上不同。的相互作用 突触核蛋白家族蛋白质与不同大脑区域的性别和年龄可能是先前观察到的性别的基础。 具体的回答。我将通过利用条件性基因敲除小鼠模型选择性地 从兴奋性神经元和不同年龄的两性试验小鼠中去除αSyn。我将研究α-Syn的影响 通过学习和记忆任务对行为进行消融，而质谱法将用于评估αSyn 可能影响蛋白质稳定性和功能的翻译后修饰。现在的技术进步 使我们能够以高空间分辨率收集同一动物的转录组学和蛋白质组学数据。 因此，我们还将确定αSyn消融对βSyn丰度和位置的影响， 以一种补偿的方式被上调。βSyn转录或翻译的区域差异可能 在突触核蛋白病的发展过程中导致特定脑区域的脆弱性。这些实验 将对αSyn的正常作用及其可能如何变化提供更深入的分子和功能了解 与性别和年龄有关。空间基因组学的应用可以为观察到的性别差异提供机制性的见解， 并为疾病诊断和治疗开辟新的途径。拟议的实验解决了以下方面的差距： 对于靶向突触核蛋白的治疗剂的翻译成功至关重要的知识，以及 提供各种技术的培训机会。

Abstract Biological variables can directly impact disease vulnerability and clinical presentation. For example, age remains the greatest risk factor for developing most neurodegenerative diseases, and while sex differences have historically been understudied, there is increasing evidence that sex can also impact disease outcomes. Alpha synuclein (αSyn) has long been implicated in the development of a multitude of neurodegenerative diseases, however, some of its normal functions remain elusive. αSyn is part of the synuclein family of proteins, a highly evolutionarily conserved group which includes beta (βSyn) and gamma synuclein. αSyn and βSyn are increasingly being studied as potential biomarkers for disease development and progression. βSyn has also been shown to be an endogenous inhibitor of αSyn aggregation, and can be upregulated in response to the removal of αSyn. Recent studies indicate that synuclein protein abundance and function could vary with sex and age. However, few studies have rigorously assessed the interaction of these biological variables in normal functioning systems. Without understanding the baseline abundance and function of these proteins, treatments targeting the synuclein family have an increased likelihood of failing in the clinic or of producing negative side effects. Results from our group have shown a sex-specific response to αSyn ablation in mice, such that female animals but not males had impaired spatial learning and memory. The goal of this proposal is to investigate the normal function of αSyn in both sexes at multiple ages. The overall hypothesis is that αSyn is transcriptionally, translationally, or functionally distinct in male and female brains. The interaction of synuclein family proteins with sex and age in different brain regions might underlie previously observed sex- specific responses. I will address the hypothesis by utilizing a conditional knockout mouse model to selectively ablate αSyn from excitatory neurons and test mice of both sexes at different ages. I will study the effects of αSyn ablation on behavior through a learning and memory task, while mass spectrometry will be used assess αSyn post-translational modifications that might impact protein stability and function. Technological advances now allow us to collect transcriptomic and proteomic data from the same animal with high spatial resolution. Therefore, we will also determine the effect of αSyn ablation on the abundance and location of βSyn, which could be upregulated in a compensatory manner. Regional differences in βSyn transcription or translation might contribute to the vulnerability of specific brain regions in the development synucleinopathies. These experiments will provide a deeper molecular and functional understanding of the normal role of αSyn and how it might change with sex and age. Utilization of spatial genomics could provide mechanistic insight into observed sex differences, and reveal new avenues for disease diagnosis and treatment. The proposed experiments address a gap in knowledge that is critical to translational success for therapeutics targeting synuclein proteins, as well as providing training opportunities in a variety of techniques.