Uncovering complex expression patterns in schizophrenia

描述（由申请人提供）：我的目标是获得先进的学科知识，这将有助于将微阵列获得的基因表达数据纳入更广泛的生物学背景。对于对基因组学感兴趣的科学家来说，理想的专业知识包括多个神经科学学科，包括（但不限于）群体遗传学，生物统计学和所研究组织的体视学评估。这种额外的专业知识将使我能够识别数据集之间的复杂关系，根据现有的连锁研究解释获得的微阵列数据，对我们的死后样本进行有限的基因分型实验，并使用无偏见的体视学采样方法验证未发现的基因表达变化。 精神分裂症患者的前额叶皮质（PFC）功能障碍与工作记忆缺陷有关，而上级颞回（STG）的功能改变与精神病有关。 此外，精神分裂症的性别表现与发病年龄、病理学、病前病史、神经影像学检查结果、药物反应性和大脑结构的差异有关。 该应用程序集中在两个关键问题：1）在不同的大脑区域内和跨不同的大脑区域是否存在精神分裂症相关的表达谱，以及2）精神分裂症相关的表达变化在性别之间是否不同？在这种情况下，我们建议测试7个特定的假设，使用3个特定的目标：目的1比较基因表达模式在12名男性受试者与精神分裂症和匹配的控制跨前额叶（PFC）和上级颞叶（STG）皮质。目标二。比较12名女性精神分裂症患者和对照组前额叶（PFC）和上级颞叶（STG）皮质的基因表达模式。目的1和2将共享相同的方法，并比较转录组：A）使用全基因组HG_U133 A和B Affymbox微阵列。B）使用定制的高灵敏度聚合物eDNA微阵列。 这些eDNA聚合物阵列，涉及我们的专有探针（专利申请正在进行中），将使我们能够对许多转录物进行改进和有针对性的评估，这些转录物太稀疏，无法通过目前可用的微阵列检测到。目标3。验证并定位微阵列未覆盖的基因表达变化至细胞类型A）在转录水平上使用原位杂交; B）在蛋白质水平上使用免疫组织化学和C）通过死后样本的基因分型跟踪最有希望的表达变化。

DESCRIPTION (provided by applicant): My objective is to acquire advanced and working knowledge of disciplines that will help place the microarray-obtained gene expression data into a broader biological context. For a scientist interested in genomics, ideal expertise encompasses multiple neuroscience disciplines, including (but not limited to) population genetics, biostatistics and stereological assessment of the studied tissue. This additional expertise will enable me to identify complex relationships between datasets, interpret the obtained microarray data in the light of existing linkage studies, perform limited genotyping experiments on our postmortem samples and verify the uncovered gene expression changes using non-biased, stereological sampling methods. Dysfunction of the prefrontal cortex (PFC) in schizophrenia has been associated with deficits of working memory, while functional changes in the superior temporal gyrus (STG) have been related to psychosis. Furthermore, the presentation of schizophrenia across genders has been associated with differences in age of onset, symptomathology, premorbid history, neuroimaging findings, drug responsiveness and brain structure. This application is focused around two critical questions: 1) Is there a schizophrenia-related expression profile within and across different brain regions and 2) Are schizophrenia-related expression changes different across genders? In this context, we propose to test seven specific hypotheses using 3 specific aims: Aim1 Compare gene expression pattern in 12 MALE subjects with schizophrenia and matched controls across the prefrontal (PFC) and superior temporal (STG) cortices. Aim 2. Compare gene expression pattern in 12 FEMALE subjects with schizophrenia and matched controls across the prefrontal (PFC) and superior temporal (STG) cortices. Aims 1 and 2 will share the same methodology, and compare the transcriptomes: A) Using whole genome HG_U133A and B Affymetrix microarrays. B) Using custom-made, high-sensitivity polymer eDNA microarrays. These eDNA polymer arrays, involving our proprietary probes (patent application in progress) will allow us an improved and targeted assessment of many transcripts that are too sparse to be detected by the currently available microarrays. Aim 3. Verify and localize the microarray-uncovered gene expression changes to cell types A) at transcript level using in situ hybridization; B) at protein level using immunohistochemistry and C) follow up the most promising expression changes with genotyping of postmortem samples.