Development of a novel biomarker test for autism risk screening

描述(由申请人提供)：自闭症谱系障碍(ASD)目前在美国大约每110名儿童中就有1名受到影响，并与家庭和社会的高经济成本有关。目前还没有一致的自闭症生物学标志物，目前的诊断是基于对行为特征和发展史的分析。对ASD最有益的治疗方法是行为疗法，如果在生命早期给予最有效的治疗。我们之前已经确定并验证了一组自闭症风险的蛋白质生物标记物 来自一大群患有自闭症儿童和未受影响的对照组母亲的妇女的血浆。这项拟议研究的主要目标是为这些生物标志物开发一种多重定量分析，以便能够在怀孕前预测生下患有自闭症的孩子的风险，特别是在那些至少有一个孩子患有自闭症并且有第二个孩子的风险增加的母亲中。这项检测也可用于产后根据孕妇自身抗体特征确定儿童是否有风险。为了实现这一目标，我们提出了以下三个具体目标：目标1：在哺乳动物和原核表达系统中表达LDH-A、LDH-B、cypin、STIP1、CRMP1和CRIMP2蛋白。评估自闭症儿童母亲的内源性自身抗体的免疫反应性，并针对这些表达的蛋白建立典型的对照。目的#2：建立LDH、cypin、STIP1和CRMP1的高通量多重检测方法。制作缓冲液和血浆中自身抗体浓度与信号的标准曲线，并评估检测的准确性、精密度和灵敏度。目的#3：使用新开发的多重分析方法对盲法血清进行的验证研究将被用来在孕期从生下自闭症孩子的母亲和生下典型发育中孩子的母亲身上采集的样本中，每种内源性自身抗体的浓度与疾病的发生相关，并以99%的可信度建立一个阳性预后的阈值。该项目将产生一种诊断性产品，让女性有机会知道她们是不是 有可能在怀孕前生下患有自闭症的孩子，从而有可能 预防措施和早期干预。

DESCRIPTION (provided by applicant): Autism spectrum disorders (ASD) currently affect about 1 in 110 children in the US, and are associated with a high economic cost to both families and society. There are currently no consistent biological markers for Autism, and diagnosis is currently based upon analysis of behavioral traits and developmental history. The most beneficial treatment for ASD is behavioral therapy, which is most effective if administered early in life. We have previously identified and validated a set of protein biomarkers for autism risk in the plasma of women from a large cohort of mothers of children with autism and unaffected controls. The main goal of the proposed research is to develop a multiplex quantitative assay for these biomarkers to be able to predict the risk of having a child with autism, prior to pregnancy, especially in those mothers who have at least one child with autism and are at elevated risk of having a second child on the spectrum. This test can also be used post-natally to determine if a child is at risk based on the maternal autoantibody profile. To achieve this goal we propose the following three Specific Aims: Aim #1: Express LDH-A, LDH-B, Cypin, STIP1, CRMP1, and CRIMP2 proteins in both a mammalian and prokaryotic expression systems. To evaluate the immunoreactivity of the endogenous autoantibodies from mothers of children with autism and typically developing controls against these expressed proteins. Aim #2: Develop high throughput multiplexed assays for LDH, Cypin, STIP1 and CRMP1. Produce standard curve of autoantibody concentration vs. signal in buffer and plasma and evaluate assay accuracy, precision, and sensitivity. Aim #3: Validation studies with blinded serum sets using the newly developed multiplex assays will be used to correlate the concentration of each of these endogenous autoantibodies to the development of disease in samples taken from mothers during pregnancy that gave birth to a child with autism vs. mothers that give birth to typically developing child, and establish a threshold for a positive prognosis with 99% confidence. The project will result in a diagnostic product that will give women an opportunity to know if they are at risk of having a child with autism prior to conception, thereby allowing the possibility to take preventative steps and early intervention.