Using Host Responses to Neurocysticercosis to Develop Novel, Brain-imaging Free Diagnostics: a US-India Partnership

利用宿主对神经囊尾蚴病的反应来开发新颖的脑成像免费诊断方法：美印合作

• 批准号: 9316725
• 负责人: Helene Carabin
• 金额: $ 51.68万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-15 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9316725
• 关键词: Antibodies; Antigens; Area; Biological; Biological Markers; Biology; Blood; Blood Tests; Brain; Brain Abscess; Brain Diseases; Brain Neoplasms; Brain imaging; Candidate Disease Gene; Cell Line; Cells; Charge; Chronic Headaches; Code; Communities; Country; Data; Detection; Diagnosis; Diagnostic; Diagnostic Imaging; Disease; Distress; Enrollment; Environment; Epilepsy; Exposure to; Faculty; Funding; Future; Gene Expression; Genes; Goals; Gold; Health; Human; Image; Imaging technology; Immune response; In Vitro; India; Infection; Institutes; Institution; Laboratory Personnel; Lead; Lesion; Mass Spectrum Analysis; Measures; Medical; Messenger RNA; Methods; Molecular; Molecular Biology Techniques; Morbidity - disease rate; Neurocysticercosis; Neurologic Symptoms; Oklahoma; Parasites; Parasitic infection; Pathway interactions; Patients; Peptides; Peripheral; Pilot Projects; Process; Proteins; RAP1A gene; Research; Resources; Science; Seizures; Sensitivity and Specificity; Serum; Serum Proteins; Specificity; Spectrometry, Mass, Electrospray Ionization; Stigmatization; Stroke; Taenia solium; Techniques; Technology; Testing; Training; Up-Regulation; Whole Blood; candidate marker; clinical diagnostics; cost; differential expression; experimental study; field study; insight; interest; intervention program; laboratory experience; link protein; medical schools; member; monocyte; mortality; neglect; nervous system disorder; novel; novel diagnostics; novel marker; overexpression; prospective; social stigma; tool; user-friendly

Neurocysticercosis (NCC) is a parasitic infection of the brain that accounts for 34% of active epilepsy cases in Vellore district, India. New methods for diagnosing NCC are badly needed because the cost of diagnostic imaging is beyond the means of most patients, and 34% of NCC patients are missed by the gold standard for antibody detection. Our National Institute of Neurological Disease and Stroke (R21NS077466) and Indian Ministry of Science and Technology (BT/MB/BRCP/06/2011)-funded pilot study used mRNA arrays of blood monocytes followed by qPCR confirmation to identify differentially expressed genes in patients with NCC- associated seizures / epilepsy (referred to as epilepsy) compared to controls with and without epilepsy. This approach identified 15 genes of interest showing highest expression levels in patients with NCC-associated epilepsy, followed by patients with resolved NCC and finally by those with idiopathic epilepsy. Expression levels of some genes differed among NCC patients with different types of brain lesions with expression decreasing as lesions resolved. In addition, sera from the same patients were analyzed by electrospray ionization mass spectrometry (ESI-MS) to identify mass/charge peaks that could discriminate between NCC patients and controls. Notably, ESI-MS also distinguished NCC from idiopathic seizures / epilepsy, and both ESI-MS and gene expression studies identified overexpression of RAP1A at the protein levels and mRNA levels, respectively, in NCC patients. These results are extremely promising for developing novel diagnostic tools. Our central hypothesis is that NCC-associated epilepsy can be diagnosed by peripheral biomarkers and will be tested with two research specific aims. Specific Aim 1 will establish the relevance of preliminarily identified candidate genes to NCC. In a first sub-aim, we will assess in-vitro relationships between exposure to T. solium metacestodes' antigens and expression of candidate genes in patient monocytes and in monocyte- like cell lines. The second sub-aim will measure candidate gene expression in monocytes prospectively in NCC patients during therapy and as their lesions change, and will be compared with expression levels of the same genes in whole blood. Specific Aim 2 will identify the proteins/peptides causing differentially expressed mass peaks in serum of patients with NCC-associated seizures and confirm their specificity to NCC. Tandem MS/MS will be used to identify proteins linked to ESI-MS mass peaks that discriminate between NCC-associated seizures and other group of patients. Capacity Building Aim 1 will train one junior faculty member and one research fellow in the use of mass spectrometry as a diagnostic tool. Capacity Building Aim 2 will train laboratory personnel at all levels in cellular and molecular techniques used to study host responses to infection. Successful completion of these aims will identify candidate biomarkers of NCC-associated seizures for field testing and determine their ability to predict different types of NCC-related lesions. Local expertise will be developed in India to conduct these studies and pursue novel insights into the biology of NCC.

神经囊肿（NCC）是大脑的寄生虫感染，占活性癫痫病例的34％ 在印度韦洛尔地区。由于诊断的成本，很难诊断NCC的新方法 成像超出了大多数患者的手段，而34％的NCC患者被金标准遗漏了 抗体检测。我们的国家神经疾病与中风研究所（R21NS077466）和印度 科学技术部（BT/MB/BRCP/06/2011）资助的试点研究使用了mrna阵列 单核细胞和QPCR确认，以鉴定NCC-患者的差异表达基因 与患有和没有癫痫的对照相比，相关的癫痫发作 /癫痫（称为癫痫）。这 方法确定了15个感兴趣的基因显示NCC相关患者的最高表达水平 癫痫病，随后是NCC解决的患者，最后是患有特发性癫痫患者。表达 NCC患者的某些基因水平不同，具有不同类型的脑病变 随着病变的解决，减小。此外，通过电喷雾分析了来自同一患者的血清 电离质谱法（ESI-MS）以识别可以区分NCC的质量/电荷峰 患者和对照。值得注意的是，ESI-MS还将NCC与特发性癫痫发作 /癫痫区区分开 ESI-MS和基因表达研究确定了Rap1a在蛋白水平和mRNA上的过表达 NCC患者的水平分别。这些结果对于开发新型诊断非常有前途 工具。我们的中心假设是，可以通过外围生物标志物诊断与NCC相关的癫痫 将通过两个研究特定目标进行测试。特定目标1将确定初步的相关性 将候选基因识别为NCC。在第一个子aim中，我们将评估暴露于暴露之间的体外关系 T.元元素的抗原和候选基因在患者单核细胞和单核细胞中的表达 喜欢细胞系。第二个子AIM将在NCC中前瞻性地测量单核细胞中的候选基因表达 治疗过程中的患者和随着病变的变化而变化，并将与同一表达水平进行比较 全血的基因。特定的目标2将识别蛋白质/肽，从而导致差异表达的质量 NCC相关性癫痫患者血清的峰值，并确认其对NCC的特异性。串联MS/MS 将用于识别与ESI-MS质量峰链接的蛋白质，这些蛋白会区分与NCC相关的蛋白质 癫痫发作和其他患者。能力建设目标1将培训一名初级教师和一名 将质谱法作为诊断工具的研究研究员。能力建筑目标2将训练 用于研究宿主对宿主反应的细胞和分子技术各级实验室人员 感染。这些目标的成功完成将确定NCC相关癫痫发作的候选生物标志物 用于现场测试并确定其预测不同类型NCC相关病变的能力。本地专业知识将 在印度开发以进行这些研究，并对NCC生物学进行新颖的见解。