Identification of glycosaminoglycans for newborn screening and therapeutic monitoring of mucopolysaccharidoses

用于新生儿筛查和粘多糖病治疗监测的糖胺聚糖鉴定

基本信息

批准号：
10010423
负责人：
Rainer Ng
金额：
$ 19.44万
依托单位：
BAEBIES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10010423
关键词：
Advocate Affect Biochemistry Biological Assay Birth Blood Blood specimen CLIA certified Categories Cessation of life Chondroitin Chondroitin Sulfates Clinical Communities Complex Data Dermatan Sulfate Development Disaccharides Disease Dose Early Diagnosis Early identification Environment Enzymes Future Genes Glycosaminoglycan Degradation Pathway Glycosaminoglycans Gold Health Heart Heparitin Sulfate Individual Inherited Joints Keratan Sulfate Laboratories Lead Legal patent Life Longevity Longitudinal Studies Measurement Measures Metabolic Diseases Methodology Methods Methylene blue Microfluidics Monitor Mucopolysaccharidoses Mucopolysaccharidosis I Mucopolysaccharidosis II Neonatal Screening Nervous system structure Neuraxis Newborn Infant Patients Performance Phase Public Health Respiratory System Running Sampling Specimen Spottings Sulfate Technology Testing Therapeutic Time Tissues Translating Treatment Efficacy Urine Whole Blood assay development base bone clinically actionable commercialization digital effective therapy enzyme deficiency enzyme replacement therapy gene panel high throughput screening innovation instrument next generation sequencing novel point of care premature rare genetic disorder response screening screening panel sugar tandem mass spectrometry technology development tool uptake

项目摘要

ABSTRACT Identification of glycosaminoglycans for newborn screening and therapeutic monitoring of mucopolysaccharidoses Mucopolysaccharidoses (MPS) are a group of 11 rare inherited metabolic diseases, each caused by a deficiency in a specific enzyme necessary for the breakdown of complex sugars termed glycosaminoglycans (GAGs). The accumulation of GAGs in various tissues causes a spectrum of health problems, including heart, bone, joint, and nervous system complications, which progressively worsen and lead to restricted mobility and premature death. Therefore, it is critical that affected newborns are identified at birth through newborn screening (NBS). However, only assays for MPS I and II are currently available in the U.S., and not all states are presently screening for MPS diseases. A methodology that can screen for MPS disorders, and identify by subtype, would be of tremendous value to the NBS community; future pilot data from a longitudinal study with such a platform would provide evidence needed to recommend uniform NBS for MPS disorders with approved therapies. The majority of currently available treatments result in a reduction in GAG accumulation and therefore monitoring of GAG levels is a crucial component of MPS treatment regimes. Standard tests for total GAG measurement are currently performed on urine samples using tandem mass spectrometry (MS/MS) or dimethyl methylene blue and have several limitations including: poor sensitivity, large sample volumes, and long turnaround times (>3 days). To overcome these challenges in NBS and therapeutic monitoring for MPS, we propose to develop a novel high-throughput digital microfluidic (DMF) platform for identification of GAGs that can be leveraged to screen for disease AND monitor disease treatment. We will use a combination of novel (patent pending) enzyme modulation assays to measure 1) total GAG levels (to identify if MPS disease is present during NBS) and 2) specific GAG categories (to determine which specific GAG is elevated during NBS). Application of targeted next generation sequencing (tNGS) as a 3rd test will determine which specific MPS/subtype is present. The GAG modulation assays will be performed on our automated DMF cartridge and platform, which can perform the GAG analyses either on dried blood spot (screening) or whole blood sample (therapeutic monitoring). The tNGS analysis will be developed in our in-house CLIA-certified laboratory. We previously demonstrated feasibility of GAG microtiter plate assays for heparan, dermatan, and keratan sulfates. We will develop assays for total GAGs and chondroitin sulfate, and translate all GAG assays to the DMF platform. The tNGS gene panel will also be developed for use during NBS. Preliminary analytical performance of the assay panel will be assessed, and a method comparison is planned against the gold standard assays to demonstrate platform equivalence. Our innovative and groundbreaking solution for identification of GAGs will dramatically increase the rate of uptake of NBS for more MPSs in state public health laboratories. The versatility of our platform to also monitor individual GAG levels during treatment will add significant value proposition during commercialization.

抽象的鉴定糖胺聚糖用于新生儿筛查和治疗监测粘多糖沉积症粘多糖贮积症 (MPS) 是一组 11 种罕见的遗传性代谢疾病，每种疾病均由缺乏粘多糖引起分解称为糖胺聚糖 (GAG) 的复合糖所必需的特定酶。这 GAG 在各种组织中的积累会导致一系列健康问题，包括心脏、骨骼、关节和神经系统并发症，逐渐恶化并导致活动受限和过早死亡。因此，通过新生儿筛查（NBS）在出生时识别受影响的新生儿至关重要。然而，目前美国仅提供 MPS I 和 II 检测方法，且并非所有州目前都在筛查 MPS 疾病。一种可以筛查 MPS 疾病并按亚型进行识别的方法是为 NBS 社区带来巨大价值；来自此类平台的纵向研究的未来试点数据将提供所需的证据，以推荐统一的 NBS 治疗 MPS 疾病和已批准的疗法。大多数目前可用的治疗方法可减少 GAG 积累，因此可监测 GAG 水平是 MPS 治疗方案的重要组成部分。目前总 GAG 测量的标准测试使用串联质谱 (MS/MS) 或二甲基亚甲蓝对尿液样本进行分析，并具有一些限制包括：灵敏度差、样本量大和周转时间长（> 3 天）。为了克服 NBS 和 MPS 治疗监测中的这些挑战，我们建议开发一种新型高通量数字微流控 (DMF) 平台，用于识别 GAG，可用于筛查疾病和监测疾病治疗。我们将采用新颖（专利待定）酶调节测定来测量 1）总 GAG 水平（以确定 MPS 疾病是否存在于 NBS）和 2）特定 GAG 类别（以确定在 NBS 期间哪个特定 GAG 升高）。应用靶向下一代测序 (tNGS) 作为第三项测试将确定存在哪种特定的 MPS/亚型。 GAG 调制测定将在我们的自动化 DMF 盒和平台上进行，该平台可以执行 GAG 对干血斑（筛查）或全血样本（治疗监测）进行分析。这 tNGS 分析将在我们内部 CLIA 认证的实验室中进行。我们之前证明了 GAG 微量滴定板测定乙酰肝素、皮肤素和角质素的可行性硫酸盐。我们将开发总 GAG 和硫酸软骨素的检测方法，并将所有 GAG 检测方法转化为 DMF平台。 tNGS 基因组也将被开发以供 NBS 期间使用。初步分析将评估检测小组的性能，并计划与黄金标准进行方法比较证明平台等效性的测定。我们的创新和突破性解决方案用于识别 GAG 将显着提高州公共卫生实验室中更多 MPS 的 NBS 吸收率。这我们平台的多功能性还可以在治疗期间监测个体 GAG 水平，这将增加显着的价值商业化期间的提议。