The sex-specific role of alpha-synuclein

摘要 生物变量可以直接影响疾病的脆弱性和临床表现。例如，年龄仍然存在 发生大多数神经退行性疾病的最大风险因素，虽然性别差异 从历史上看，研究不足，越来越多的证据表明，性行为也会影响疾病的结局。Alpha 突触核蛋白(αsyn)长期以来一直与多种神经退行性疾病的发生有关， 然而，它的一些正常功能仍然难以捉摸。αSyn是突触核蛋白家族的一员，是一种高度 进化上保守的群体，包括β(β，突触核蛋白)和伽马突触核蛋白。α系统和β系统是 作为疾病发展和进展的潜在生物标志物，越来越多的人被研究。βSYN也有 已被证明是αSYN聚集的内源性抑制物，并可在响应 删除α系统。最近的研究表明，突触核蛋白的丰度和功能可能会因性别和 年龄。然而，很少有研究严格评估这些生物变量在正常人群中的相互作用。 正常运行的系统。如果不了解这些蛋白质的基线丰度和功能，治疗 以突触核蛋白家族为靶点在临床上失败或产生负面影响的可能性增加 效果。我们小组的结果显示，小鼠对α突触的消融具有性别特异性反应，因此雌性 动物，而不是雄性，空间学习和记忆受损。这项提案的目标是调查 α同步蛋白在不同年龄段男女间均有正常功能。总体假设是α系统是 男女大脑在转录、翻译或功能上不同的男性和女性大脑。的相互作用 性别和年龄在不同脑区的突触核蛋白家族蛋白可能是先前观察到的性别的基础。 具体的回应。我将通过利用条件性基因敲除小鼠模型来解决这一假设 从兴奋性神经元中消融α突触，并测试不同年龄的雌雄小鼠。我将研究α系统的影响 通过学习和记忆任务对行为进行消融，同时将使用质谱仪评估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和蛋白质组数据。 因此，我们还将确定α突触消融对β突触的丰度和位置的影响，这可能 以补偿的方式被上调。βSYN转录或翻译的地区差异可能 导致发育中的联核症的特定脑区的脆弱性。这些实验 将提供更深入的分子和功能方面的了解，以了解α系统的正常作用以及它可能如何改变 性别和年龄。利用空间基因组学可以提供对观察到的性别差异的机械性洞察， 并揭示了疾病诊断和治疗的新途径。拟议中的实验解决了一个缺口 对于以突触核蛋白为靶点的疗法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.