Cellular Phenotypes of Genetic Variants in Mucopolysaccharidosis

摘要 目前迫切需要发展系统的方法来评估基因组瓦里的功能， 蚂蚁临床基因组检测正在迅速发展，随之而来的是比以往任何时候都多的遗传变异， 不能被定义为致病或良性。这个问题影响到临床医生和他们的病人， 努力理解和解释分子诊断报告，结果的含义，以及如何 在缺乏确切信息的情况下管理他们的病人。涉及单细胞产生 高内容、高分辨率的功能数据对于解决由变化引起的问题至关重要， 不确定的重要性。在这里，我们建议利用一种新的基于细胞的平台，使用机器学习- 以确定定义致病性的形态表型的组合。我们将应用 技术的综合功能评估的变体在IDS，基因负责猎人 综合征该提案中概述的核心假设是，测量直接函数的实验数据- 变异的典型效应将为准确的疾病风险预测提供信息。此外，我们假设，在体外， 基于形态特征的基于细胞的测定与现有的基于细胞的测定相比将更准确地检测疾病。 使用人造基质进行生化测试。我们将在变体文库中使用 在Buchser实验室的RaftSeq管道中，将建立细胞表型，然后对我们进行测试- 第二组变体与拯救实验相结合。这些结果将为变异分类提供信息， IDS分子检测和改善个体的诊断，包括通过低艾杜糖醛酸-2-磺胺酸鉴定的个体。 新生儿筛查中的酶活性。

ABSTRACT There is presently an urgent need to develop methodical approaches to evaluate the function of genomic vari- ants. Clinical genomic testing is growing rapidly, and with it a larger-than-ever number of genetic variants that cannot be defined as either disease-causing or benign. The problem affects clinicians and their patients, who struggle to understand and interpret molecular diagnostic reports, the implications of the results, and how to manage their patients in the absence of definitive information. Systems that involve single cells to generate high-content, high-resolution functional data are of paramount importance to solving the problem posed by var- iants of uncertain significance. Here, we propose to utilize a novel cell-based platform that uses machine learn- ing to determine the combination of morphological phenotypes that define pathogenicity. We will apply the technology to the comprehensive functional assessment of variants in IDS, the gene responsible for Hunter syndrome. The core hypothesis outlined in this proposal is that experimental data measuring the direct func- tional effects of variants will inform accurate disease risk prediction. In addition, we hypothesize that an in vitro, cell-based assay based on morphological features will more accurately detect disease compared to existing biochemical testing using artificial substrates. We will perform a functional assay in a variant library using the RaftSeq pipeline in the Buchser laboratory, where a cellular phenotype will be established and then tested us- ing a second set of variants combined with rescue experiments. The results will inform variant classification in IDS molecular testing and improve diagnosis of individuals including those identified by low iduronate-2-sulfa- tase activity on newborn screening.