An acoustofluidic avidity cytometer for massive parallel profiling single autoreactive T cell in autoimmune disease

项目摘要 自身反应性T细胞反应针对中枢神经系统衍生的自身抗原，并导致几种 脱髓鞘自身免疫性疾病，包括多发性硬化症、急性播散性脑脊髓炎和 视神经脊髓炎。多发性硬化症，这些疾病中最常见的一种，影响着200多万人 全世界约有40万人在美国，是一种慢性炎症性疾病，导致 脑和脊髓出现脱髓鞘病变和斑块。然而，目前的临床测试都不能 信心十足地预测临床多发性硬化症的进展或治疗效果 敏感有效的生物标志物。 多发性硬化症患者的T细胞表现为活化的表型，与髓鞘蛋白的亲和力增加。 因此，我们假设自身反应性T细胞的亲和力是一种疾病标记物，可以用来监测 疾病进展，判断治疗反应，或发现新的生化疾病标记物 硬化症。然而，目前的技术都没有实现高灵敏度和高通量 在临床样本中，在单细胞水平上测量细胞亲和力。PI在工程上的进展 “声流体学”，允许以意想不到的分辨率精确操纵单细胞和液体，显示 在测量高度异质性的临床自身反应性T细胞亲和力以及 与PI的高吞吐量相结合时，可能会对感兴趣的单个单元格进行排序和分析 微流控技术。 目的是创造一种声流亲和力细胞仪，以克服临床检测的障碍。 自身免疫性疾病，如多发性硬化症。然而，有几个重要的障碍，包括 建立一种高灵敏度和高通量亲和力细胞仪来分析单个自身反应性髓鞘特异性T细胞 并测定其亲和力在临床标本中的分布。结合高通量微流控技术将 允许筛选感兴趣的单个自身反应性T细胞以寻找新的自身抗原。建议的工具 可能会给目前的临床测试带来革命性的变化，并发现新的诊断、预后和 自身免疫性疾病的治疗效果，包括多发性硬化症、类风湿性关节炎、克罗恩病等。

Project Summary Autoreactive T cell responses target central nervous system-derived autoantigens and cause several demyelinating autoimmune diseases including multiple sclerosis, acute disseminated encephalomyelitis, and neuromyelitis optica. Multiple sclerosis, the most common of these diseases affecting over two million people worldwide and approximately 400,000 people in United States, is a chronic inflammatory disease, causing lesions and plaques of demyelination in the brain and spinal cord. However, none of the current clinical tests can confidently predict clinical multiple sclerosis progression or treatment efficacy due to a lack of sufficiently sensitive and effective biomarkers. T cells in multiple sclerosis patients display an activated phenotype with an increased avidity to myelin protein. As a result, we hypothesize that the avidity of autoreactive T cells is a disease marker that can be used to monitor disease progress, judge therapeutic response, or discover new biochemical disease markers for multiple sclerosis. However, none of the current technologies have achieved high-sensitivity and high-throughput measurement of cell avidity, in clinical samples, at the single cell level. The PI’s engineering advances in ‘Acoustofluidics,’ allowing for precise manipulation of single cells and liquids at unexpected resolution, shows promising potential for measuring the avidity of highly heterogeneous, clinical autoreactive T cells as well as for potentially sorting and analyzing single cell of interest, when combined with the PI’s ‘High Throughput Microfluidic’ technology. The objective is to create an acoustofluidic avidity cytometer to overcome the barriers in detecting clinical autoimmune disorders such as multiple sclerosis. However, there are several significant hurdles which include building a high-sensitivity and high-throughput avidity cytometer to profile single autoreactive myelin-specific T cells and determine their avidity distribution in clinical samples. Incorporating high-throughput microfluidics will allow the screening of single autoreactive T cells of interest for new autoantigens. The proposed instrumentation will potentially revolutionize the current clinical testing and discovery new markers for diagnosis, prognosis, and treatment efficacy of autoimmune disease including multiple sclerosis, rheumatoid arthritis, Crohn’s disease, etc.