Development of an Autism Spectrum Disorder Screening Test based upon Metabolic Profiling of Fresh Blood Samples

自闭症谱系障碍(Asd)是一种神经发育障碍，其特征是显著的相互作用缺陷。 社交互动、沟通障碍和受限、重复的行为，患病率为#年每68名儿童中就有1名 美国。早期干预计划已被证明在提高智商、语言和社交技能方面有效 18个月大的蹒跚学步的孩子。ASD的诊断目前仅基于临床和父母的观察， 作为一种基于实验室的ASD诊断试验，目前尚不存在基于稳健生物标志物的ASD诊断方法。我们之前的团队 证明来自ASD患者的淋巴母细胞系(LCL)在以下情况下产生代谢活性降低 与典型的发展中(TD)对照相比，氨基酸色氨酸是唯一可用的能量来源。在最近一次 使用Bootstrap聚集法对来自1-10岁个体的50个ASD和50个TD细胞系的分类进行评估 基于色氨酸和一系列扩展离子的方法能够正确地对单个细胞系进行分类 相对于临床诊断，灵敏度和特异度均为92%。该分类器表现出了较好的性能。 较年轻的年龄组，特别是1至3岁的人，这也是筛查的理想目标范围 ASD.基于这些专利方法的实验室生物标记物测试的开发将有可能成为第二个 初步行为观察后的分级客观测试已确定儿童有患自闭症的风险，因此 为诊断的确认和早期治疗提供了条件。潜在地，这样的测试也将(1)导致更好的 了解这种疾病的致病机制，(2)帮助鉴定生化 描绘ASD亚组的签名，以及(3)突出改进和个体化治疗的新靶点 接近了。然而，现有测试方法的广泛使用目前受到成本和时间的限制 与为每个样本产生永生化细胞系相关联，或者通过可以 从儿童那里获得，这是获得足够数量的活细胞以进行代谢谱分析所必需的。 因此，拟议工作的目标是开发方法和技术来对 从少量新鲜血液中提取免疫细胞，以实现ASD的可行筛查测试。努力将首先集中在 关于提高当前活细胞代谢图谱技术的灵敏度。分析平台的小型化 然后将实施以减少每个测试条件所需的单元数量，从而限制必要的 血样体积。至少100名被诊断为ASD的患者和100名TD对照组的血液样本将 根据制定的方法和方案，以盲法进行代谢分析。的表现 然后将对多个分类器进行评估，以确定这些技术识别诊断为 ASD.这项工作的成功完成将直接导致第二阶段调查，这将涉及测试 扩大队列，包括其他神经发育障碍，并最终确定商业上可行的测试设计 服务、平台或套件。拟议技术的商业化最终将提供一项筛查测试，以 识别患有ASD风险增加的幼儿，从而提供更早的诊断和改善的行为结果。

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by significant deficits in reciprocal social interactions, impaired communication and restricted, repetitive behaviors, with a prevalence of 1 in 68 children in the US. Early intervention programs have been shown to be effective for improving IQ, language, and social skills for toddlers as young as 18 months of age. Diagnosis of ASD is currently based solely on clinical and parental observations, as a laboratory-based diagnostic test for ASD based on robust biomarkers does not exist currently. Our team previously demonstrated that lymphoblastoid cell lines (LCLs) from individuals with ASD generate reduced metabolic activity when the amino acid tryptophan is the only available energy source, relative to typically developing (TD) controls. In a recent evaluation of 50 ASD and 50 TD cell lines from individuals aged 1 to 10, a classifier utilizing a Bootstrap aggregation approach based upon tryptophan and an expanded series of ions was able to properly classify individual cell lines with both a sensitivity and specificity of 92% relative to the clinical diagnosis. The classifier showed better performance in the younger age groups, particularly in individuals aged 1 to 3, which is also the ideal target range for a screening test for ASD. The development of a laboratory biomarker test based on these patented methods will potentially serve as a second tier objective test following initial behavioral observations have identified a child at risk of developing ASD, thereby providing for confirmation of the diagnosis and earlier treatment. Potentially, such a test will also (1) lead to a better understanding of the pathogenic mechanisms underlying this condition, (2) assist in the identification of biochemical signatures delineating ASD sub-groups, and (3) highlight novel targets for improved and individualized treatment approaches. Widespread use of the existing test methods, however, is currently limited either by the cost and time associated with generating immortalized cell lines for each sample or by the size of the fresh blood sample that can be acquired from children, which is necessary to obtain sufficient numbers of live cells for the metabolic profiling assay. Therefore, the objective of the proposed effort is to develop methods and technologies to perform metabolic profiling on immune cells from small volumes of fresh blood to enable a viable screening test for ASD. Efforts will first be focused on improving the sensitivity of the current live-cell metabolic profiling techniques. Miniaturization of the assay platform will then be implemented to reduce the number of cells needed for each test condition, thereby limiting the necessary blood sample volume. Blood samples from a minimum of 100 subjects diagnosed with ASD and 100 TD controls will undergo metabolic profiling in a blinded fashion according to the methods and protocols developed. The performance of multiple classifiers will then be evaluated to determine the ability of the techniques to identify individuals diagnosed with ASD. The successful completion of this effort will lead directly to the Phase 2 investigation, which will involve testing an expanded cohort, including other neurodevelopmental disorders, and finalizing the design of a commercially viable testing service, platform, or kit. Commercialization of the technology proposed will ultimately provide for a screening test to identify toddlers at increased risk for ASD, thereby providing for earlier diagnosis and improved behavioral outcomes.