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.

摘要 糖胺聚糖在新生儿筛查和治疗监测中的应用 粘多糖 粘多糖沉积症（MPS）是一组11种罕见的遗传性代谢疾病，每种疾病都是由缺乏引起的。 在一种特殊的酶中，这种酶是分解称为糖胺聚糖（GAG）的复合糖所必需的。的 GAG在各种组织中的积累引起一系列健康问题，包括心脏、骨骼、关节和 神经系统并发症，逐渐恶化，导致行动受限和过早死亡。 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，在这方面， 目前在美国仅可获得MPS I和II的分析，目前并不是所有的州都在筛查 MPS疾病。一种可以筛查MPS疾病并通过亚型识别的方法， 对国家统计局社区的巨大价值;未来的试点数据，从纵向研究与这样一个平台将 提供所需证据，以推荐采用获批治疗的MPS疾病的统一NBS。大多数 目前可用的治疗导致GAG蓄积减少，因此监测GAG 水平是MPS治疗方案的关键组成部分。目前，用于总GAG测量的标准测试是 使用串联质谱法（MS/MS）或二甲基亚甲蓝对尿样进行分析， 存在一些局限性，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，用于鉴定可以 用于筛查疾病和监测疾病治疗。我们将使用一种新颖的（专利） 酶调节试验，以测量1）总GAG水平（以确定在治疗期间是否存在MPS疾病） NBS）和2）特异性GAG类别（以确定哪种特异性GAG在NBS期间升高）。的应用 靶向下一代测序（tNGS）作为第三次测试将确定存在哪种特定MPS/亚型。 GAG调节试验将在我们的自动DMF检测盒和平台上进行， 对干血斑（筛选）或全血样本（治疗监测）进行GAG分析。的 tNGS分析将在我们内部CLIA认证的实验室进行。 我们以前证明了乙酰肝素、皮肤素和角质素的GAG微量滴定板测定法的可行性 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。还将开发tNGS基因面板以供NBS期间使用。初步分析 将评估检测试剂盒的性能，并计划与金标准进行方法比较 用于证明平台等效性的试验。我们的创新和突破性的解决方案， GAGs将大大提高国家公共卫生实验室中更多MPS的NBS吸收率。的 我们的平台在治疗期间还可监测个体GAG水平的多功能性将显著增加价值 在商业化过程中。