Accessible FSHD diagnostics through epigenetic analysis

摘要 摘要面肩臂肌营养不良症(FSHD)是一种常染色体显性遗传性肌肉疾病。 这影响了所有年龄段的男性和女性。FSHD被认为是第三种最流行的疾病 肌营养不良(约12：100,000成人)；然而，由于不同的严重程度和并发症 通过基因检测，许多人仍然没有得到诊断，患病率可能会很高 再高一点。神经科医生对FSHD的临床诊断可能会因FSHD的变异性而复杂化。 疾病表现，常被误诊为肢体带状型肌营养不良症或 简单的慢性损伤。使用基因特异性对神经肌肉疾病进行标准基因检测 靶向测序小组或甚至整个外显子组测序不能识别FSHD1 (代表FSHD病例的95%)，很容易漏掉FSHD2(&lt；5%的病例)。跟随 在临床诊断方面，基因检测费用昂贵，许多人无法获得。因此， 由于起病晚，虚弱进展缓慢，临床变数大 陈述，以及缺乏获得或了解特定基因检测的机会，许多FSHD 个人多年来仍未得到诊断，这种疾病在家庭中传播，影响 跨越几代人的多个成员。此外，由于目前的基因检测 FSHD昂贵、耗时、复杂、不完整，并且与标准基因不兼容 对于其他神经肌肉疾病的检测技术，我们对其患病率知之甚少 FSHD在世界许多地区，以及在某些族裔和服务不足的人群中，甚至 在美国国内。为了满足这一需求，我们最近开发了一种适用于所有形式的新型诊断测试 基于疾病基因座的表观遗传状态的FSHD。这种分析可以被执行 来自任何来源的基因组DNA，包括唾液，以及几乎任何质量的，从而使它 任何年龄的家庭成员和世界各地的人都可以广泛使用和访问 服务不足的社区。在这里，我们将把当前的诊断协议从低- 吞吐量单次使用，以高吞吐量的下一代测序协议。这一过渡 将大大降低化验成本，并使其更容易满足CLIA的批准。 此外，我们正在与MOVE FSHD研究合作，并进行表观遗传学分析 所有250名参与者的身份。这些受试者将进行深入的临床评估并记录在案 自然病史，这将使我们能够确定疾病基因座的表观遗传学特征是否 可作为FSHD的预后生物标志物。

SUMMARY Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant muscle disease that affects males and females of all ages. FSHD is thought to be the third most prevalent muscular dystrophy (~12:100,000 adults); however, due to variable severity and complications with genetic testing, many people remain undiagnosed and the prevalence may be significantly higher. The clinical diagnosis of FSHD by a neurologist can be complicated by the variability of disease presentation, and is often misdiagnosed as a limb-girdle type muscular dystrophy or a simple chronic injury. Standard genetic testing for neuromuscular disease using gene-specific targeted sequencing panels or even whole exome sequencing does not identify FSHD1 (representing >95% of FSHD cases) and can easily miss FSHD2 (<5% of cases). Following clinical diagnosis, genetic testing is expensive and not accessible to many populations. Thus, due to a combination of late onset, slow progression of weakness, highly variable clinical presentation, and lack of access to or knowledge of specific genetic testing, many FSHD individuals remain undiagnosed for years and the disease runs through families, affecting multiple members across several generations. In addition, since current genetic testing for FSHD is costly, time-consuming, complex, incomplete, and incompatible with standard genetic testing techniques for other neuromuscular diseases, we know very little about the prevalence of FSHD in many parts of the world as well as in certain ethnic and underserved populations even within the US. To address this need, we recently developed a novel diagnostic test for all forms of FSHD based on the epigenetic status of the disease locus. This analysis can be performed on genomic DNA from any source, including saliva, and of almost any quality, thereby making it widely available and accessible to family members of any age and people around the world in underserved communities. Here, we will convert the current diagnostic protocol from low- throughput single use to a high-throughput next-generation sequencing protocol. This transition will greatly reduce the cost of the assay and render it more amenable to meeting CLIA approval. In addition, we are collaborating with the MOVE FSHD study and performing epigenetic analysis on all 250 participants. These subjects will have in-depth clinical evaluations and documented natural histories which will allow us to determine if epigenetic profiles of the disease locus can serve as a prognostic biomarker for FSHD.