Facilitating personalized medicine of monogenic stone patients by genetic characterization

利用基因特征促进单基因结石患者的个体化用药 下一代测序(NGS)，包括使用已知和候选的目标(T)NGS小组 基因，强调了严重的遗传性尿路结石比 之前受到重视，现在有大约40个基因牵涉其中。这些单一货币的美元走在了 个体化用药在肾脏病中的应用。治疗可以针对酶/mRNAs编码关键字 缺陷代谢途径中导致有害中间体积累或通过伴侣积累的步骤 帮助折叠和正确运输突变蛋白质的治疗。基因和等位基因信息日益成为 参与这些有针对性的临床试验的先决条件。在过去的10年里，罕见的肾结石 联合会(RKSC)专注于招募和表征具有单基因美元的患者群体， 其目的是了解疾病的自然历史，进行基因/表型研究，以及 对病人进行分类，以便进行临床研究。这最初涉及Sanger测序，但最近TNG带有 ~100基因面板。作为这项筛查的一部分，被推定患有原发性高草酸尿症(PH)或 齿状神经病(两种常见的美元)，但桑格的这些疾病的规范基因没有突变 测序，在面板上进行筛选。在297名分析的患者中，发现了一种单基因原因 30例(10.1%)，发现11个不同的基因突变。在过去的一年里，所有的RKSC都招募了患者 已经使用这种TNGs方法进行了初步的基因筛查，在102个家庭中筛查了33个 (32.4%)已鉴定出8个不同的基因。有趣的是，在约15%的案例中 遗传复杂性与第二个单基因中的另一个可能的致病变异相一致。 不幸的是，RKSC的资金最近丢失了，但我们计划保持财团的完好无损 建议对通过RKSC组招募的单基因UD进行遗传学表征，主要是为了识别 纳入临床试验的患者。这项建议有两个具体目标：1.携带A基因的患者 全球NGS的表型与单基因形式的肾结石或肾钙沉积症一致 方法和2.表征和分析在致病基因之外的变异阵列 单基因结石患者。该项目有联合PIs，其中一位是美国大学临床和生化方面的专家 其中一位是单基因肾脏疾病遗传学专家(哈里斯博士)。测试的TNG 将与开发和验证的生物信息学管道一起使用，以确定可能的 致病变种。这些结果将返回给转诊医生，并提供咨询和保障 不同的构象，提供诊断信息，允许个性化治疗，并鼓励临床 试训。为研究而积累的关于基因变异和变异组合的知识 将有助于更好地了解这些疾病，并提供对引起这些疾病的遗传因素的见解 表型修饰。

Facilitating personalized medicine of monogenic stone patients by genetic characterization Next generation sequencing (NGS), including employing a targeted (t)NGS panel of known and candidate genes, is highlighting that severe, inherited forms of urinary stone disease (USD) are more common than previously appreciated, with ~40 genes now implicated. These monogenic USD are at the forefront of the application of personalized medicine in nephrology. Treatments can target enzymes/mRNAs encoding key steps in the defective metabolic pathway that lead to accumulation of harmful intermediates or via chaperone treatments to help fold and correctly traffic the mutated proteins. Genic and allelic information is increasingly a prerequisite for involvement in these targeted clinical trials. Over the past 10 years, the Rare Kidney Stone Consortium (RKSC) has focused on recruiting and characterizing patient populations with monogenic USD with the aims to understand the natural history of the disorders, conduct genotype/phenotype studies, and categorize patients for clinical studies. This initially involved Sanger sequencing but more recently tNGS with a ~100 gene panel. As part of this screening, patients presumed to have either primary hyperoxaluria (PH) or Dent disease (two common USD), but without mutations in the canonical genes for these disorders by Sanger sequencing, were screened on the panel. Out of 297 analyzed patients, a monogenic cause was detected in 30 cases (10.1%), with mutations identified in 11 different genes. In the past year, all RKSC recruited patients have been primarily genetically screened employing this tNGS approach and of 102 families screened 33 (32.4%) have been genetically resolved with 8 different genes identified. Interestingly, in ~15% of cases genetic complexity was identified with an additional likely pathogenic variant in a second monogenic gene. Unfortunately, funding for the RKSC was recently lost but we plan to keep the consortium intact and here propose to genetically characterize monogenic USD recruited through the RKSC groups, primarily to identify patients for inclusion in clinical trials. The proposal has two specific aims: 1. Genotype patients with a phenotype consistent with a monogenic form of nephrolithiasis or nephrocalcinosis using global NGS approaches and 2. Characterize and analyze the array of variants beyond the causative gene in monogenic stone patients. The project has co-PIs, one an expert in clinical and biochemical aspects of USD (Dr. Lieske), and one an expert in the genetics of monogenic kidney diseases (Dr. Harris). Tested tNGS approached will be employed along with a developed and proven bioinformatics pipeline to identify likely causative variants. These results will be returned to the referring physician, with safeguards of counseling and variant conformation, to provide diagnostic information, to allow personalized treatment, and encourage clinical trial recruitment. The accumulated knowledge of gene variants and variant combinations derived for the study will allow for better understanding of these diseases and provide insights into genetic factors causing phenotypic modification.