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

• 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
• 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

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 粘二糖 粘多糖糖（MPS）是11种稀有遗传代谢疾病的组，每种疾病是由缺陷引起的 在分解的称为糖胺聚糖（GAGS）的复杂糖分所需的特定酶中。这 各种组织中插科打的积累会导致许多健康问题，包括心脏，骨骼，关节和 神经系统并发症，逐渐恶化并导致流动性限制和过早死亡。 因此，至关重要的是，通过新生儿筛查（NBS）在出生时发现受影响的新生儿。然而， 目前仅在美国提供国会议员I和II的测定法，目前并非所有州都在筛选 国会议员疾病。可以筛选MPS疾病并通过亚型识别的方法将是 对NBS社区的巨大价值；通过这样一个平台的纵向研究的未来试验数据将 提供所需的证据，以推荐具有批准疗法的国会议员疾病的统一NB。多数 当前可用的治疗方法导致堵嘴积累减少，因此监测了堵嘴 水平是MPS治疗方案的关键组成部分。目前正在进行总插科打测量的标准测试 使用串联质谱法（MS/MS）或二甲基甲基蓝色在尿液样品上进行 几个局限性，包括：灵敏度不佳，样品量较大和较长的周转时间（> 3天）。 为了克服NB中的这些挑战和对国会议员的治疗监测，我们建议开发一个 新型的高通量数字微流体（DMF）平台，用于识别可能是 利用筛查疾病和监测疾病治疗。我们将使用小说的组合（专利 酶调节测定法进行测量1）总GAG水平（确定是否存在MPS疾病 nbs）和2）特定的插入类别（确定在NBS期间哪个特定插入率升高）。应用 针对的下一代测序（TNG）作为第三个测试将确定存在哪些特定的MPS/亚型。 插入调节测定法将在我们的自动DMF墨盒和平台上执行，可以执行 GAG在干血点（筛查）或全血液样本（治疗性监测）上分析。这 TNGS分析将在我们的内部CLIA认证实验室中进行开发。 我们以前证明了heparan，Dermatan和Keratan的GAG微量滴定板测定的可行性 硫酸盐。我们将开发总堵嘴和硫酸软骨素的测定法，并将所有插科打过分析转换为 DMF平台。 TNGS基因面板也将在NBS期间开发用于使用。初步分析 将评估测定面板的性能，并计划与黄金标准进行比较 证明平台等效的测定法。我们的创新和开创性解决方案，以识别 在州公共卫生实验室中，插科打s将大大提高NBS的吸收率。这 我们平台的多功能性也可以在治疗过程中监视单个插科打液水平将增加重要的价值 商业化期间的命题。