Modelling the molecular pathogenesis of triple A syndrome (AAAS) with iPSC-derived neurons and adrenocortical cells.

氧化应激（OS）发生在身体的天然抗氧化剂防御不能保护免受自由基的损害时，自由基是由身体的基本过程产生的。OS是许多退行性疾病的基础，包括三A综合征（AAAS）。AAAS是一种罕见的遗传性疾病，出现在儿童早期。它会导致进行性的，使人衰弱的问题，包括肾上腺衰竭，如果不治疗会危及生命，以及神经系统的严重退化。AAAS的治疗旨在缓解症状，而不是防止对生活质量或早期死亡的破坏性影响。AAAS由AAAS基因突变引起，该基因负责ALADIN的产生，ALADIN是一种知之甚少的蛋白质，被认为对许多基本细胞过程至关重要。详细了解该基因突变如何导致AAAS对于帮助开发治疗方法并可能减缓肾上腺和神经系统损伤是必要的。这一领域的研究对其他肾上腺和神经系统退行性疾病也有更广泛的影响。现有的用于研究AAAS的疾病模型都存在明显的缺陷。从病人身上获取肾上腺或脑组织是不可行的。大多数研究都是使用从患者皮肤中获得的细胞进行的，这些细胞在AAAS中通常不会受到影响。其他研究使用的癌细胞与正常细胞和患者细胞有显著差异。AAAS动物模型没有表现出该疾病的典型特征。因此，我与桑格研究所合作，开发了基因编辑的AAAS人类干细胞模型。这些细胞将用于产生AAAS肾上腺和脑组织细胞，为研究主要受疾病影响的组织中的AAAS提供接近自然的条件。这些新型疾病模型将用于揭示AAAS中异常的细胞过程和OS驱动因素。通过一系列严格的实验，我将研究AAAS如何影响肾上腺和脑组织的正常功能，以及疾病过程中的异常情况。我将使用RNA测序，这是一种用于检测AAAS疾病导致的基因表达变化的方法。通过检查患病细胞与健康细胞中基因的活性，我将能够确定可归因于AAAS和ALADIN在细胞内的关键作用的关键差异。另一种假设是AAAS基因的突变导致细胞能量发生器（称为线粒体）出现问题。如果我的实验支持这个理论，我将进行实验来详细检验。最终，我的目标是确定潜在的治疗靶点，可以有针对性地减缓或停止AAAS和其他退行性疾病的疾病进展。肾上腺干细胞建模是一个新的研究领域，该项目收集的信息将进一步推动这一领域的发展，从而将这项技术用于治疗肾上腺衰竭患者。

Oxidative stress (OS) occurs when the body's natural antioxidant defences fail to protect against damage from free radicals, produced by essential processes in the body. OS underlies numerous degenerative conditions, including Triple A syndrome (AAAS). AAAS is a rare genetic disorder, which presents in early childhood. It results in progressive, debilitating problems including adrenal gland failure, which is life threatening if untreated, and severe degeneration of the nervous system. Treatments for AAAS are aimed at symptomatic relief rather than preventing the devastating impact on quality of life or early death. AAAS results from mutations in the AAAS gene, responsible for the production of ALADIN, a poorly understood protein thought to be critical for many essential cellular processes. A detailed understanding of how mutations in this gene cause AAAS is necessary to aid the development of treatments and potentially slow the adrenal and nervous system damage. Research in this field also has wider implications for other adrenal and degenerative disorders of the nervous system. The existing disease models used to study AAAS all have significant shortcomings. Obtaining adrenal or brain tissue from patients is not feasible. Most research is performed using cells obtained from patients' skin, which is not typically affected in AAAS. Other studies use cancer cells, which differ significantly from normal and patient cells. The AAAS animal model does not exhibit the classical features of the disease. Therefore in collaboration with the Sanger Institute, I have developed gene-edited human stem-cell models of AAAS. These will be used to generate AAAS adrenal and brain tissue cells, providing near-natural conditions to study AAAS in the tissues primarily affected by the disease.These novel disease models will be used to uncover the abnormal cell processes and drivers of OS in AAAS. Through a rigorous series of experiments, I will investigate how AAAS affects the normal functioning of adrenal and brain tissue and what abnormalities underlie the disease process. I will use RNA sequencing , a method used to examine any changes in gene expression that result from AAAS disease. By examining the activity of genes in diseased compared to healthy cells, I will be able to identify key difference that can be attributed to AAAS and signpost ALADIN's critical roles within the cell. Another hypothesis is that mutations in the AAAS gene cause problems in the cell's energy generators, called mitochondria. If my experiments support this theory, I will undertake experiments to examine this in detail. Ultimately, I aim to identify potential therapeutic targets that could be targeted to slow or halt disease progression in AAAS and other degenerative diseases. Adrenal stem cell modelling is a new field of research and information gathered by this project will further advance this field, leading to the use of this technology as a treatment for patients with adrenal failure.