Mediators of Lithium Action in Olfactory Epithelium

嗅觉上皮细胞中锂作用的介质

• 批准号: 8456186
• 负责人: Evaristus A Nwulia
• 金额: $ 53.87万
• 依托单位: HOWARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-17 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8456186
• 关键词: Acute; Adult; Aftercare; Age; Antibodies; Autopsy; BCL2 gene; Biological Markers; Biopsy; Bipolar Disorder; Blood; Blood Cells; Brain; Brain-Derived Neurotrophic Factor; Candidate Disease Gene; Cells; Chronic; Clinical; Controlled Study; Data; Development; Disease; Disease Marker; Economic Burden; Functional disorder; Future; Gender; Gene Expression; Gene Mutation; Genes; Glycogen Synthase Kinase 3; Goals; Growth Associated Protein 43; Investigation; Life; Literature; Lithium; Measures; Mediating; Mediator of activation protein; Messenger RNA; Molecular; Molecular Profiling; Moods; NGFR Protein; Natural regeneration; Nerve Tissue; Neuraxis; Neurons; Nose; Olfactory Epithelium; Olfactory Receptor Neurons; Participant; Patients; Pattern; Pharmaceutical Preparations; Pilot Projects; Probability; Proliferating; Prospective Studies; Protein Kinase C; Proteins; Proto-Oncogene Proteins c-akt; Race; Recruitment Activity; Relative (related person); Resources; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Scientist; Severities; Smell Perception; Smoking; Societies; Structure; Surrogate Markers; Symptoms; Techniques; Testing; Therapeutic; Therapeutic Effect; Tissues; Translations; base; brain tissue; burden of illness; cell type; cohort; density; disorder control; gene function; human IGFBP2 protein; improved; indexing; inositol 1-phosphate; interest; laser capture microdissection; lymphoblast; molecular marker; myo-Inositol-1-Phosphate Synthase; myo-inositol-1 (or 4)-monophosphatase; myristoylated alanine-rich C kinase substrate; neurogenesis; neuronal survival; non-smoking; novel; olfactory marker protein; peripheral blood; pre-clinical; preclinical study; preempt; prognostic; response; success; therapeutic target

DESCRIPTION (provided by applicant): Bipolar Disorder (BD) is a chronic debilitating illness with substantial societal burden. A major impediment in our ability to develop improved therapeutics for BD is the fact that our understanding of its pathophysiology is incomplete. Lithium, used for treatment of BD, alters steady-state mRNA levels of a large number of genes; notably GSK-32, inositol monophosphatase (IMPase), MIP Synthase, Bcl-2, Akt, Protein Kinase C, MARCKS, insulin-like growth factor binding protein-2 (IGFBP-2), and ERK. But it remains unknown, how any of these molecular changes bring about clinical response, if, at all. Hitherto, studying cellular and molecular effects of lithium in the central nervous (CNS) tissues of living BD patients has been infeasible. Although postmortem brains have been useful for studies of biomarkers, they are insufficient to capture state-dependent molecular signature of BD. Blood cells may not reflect adequately, molecular disposition in neuronal cells. In this study, we propose use of olfactory epithelium (OE) via nasal biopsy, combined with novel laser-captured microdissection to elucidate molecular and cellular markers of prospectively-determined clinical response to lithium. This will be followed by determination of parallel dispositions of these molecular patterns in lymphoblasts obtained from same subjects, for possible clinical and prognostic utility of blood markers. OE is a unique part of the CNS that continually regenerates and differentiates into mature olfactory receptor neurons (ORNs) throughout life. This presents an avenue to examine the neurodevelopmental hypotheses of BD through examination of changes in the densities of proliferating (p75NGRF+) neuronal precursors and mature (olfactory marker protein+) neurons (surrogate markers of neurogenesis and neuronal survival in OE). Also of particular interest, dysregulation of genes relevant to lithium action, and therefore BD may be reflected in deficits in olfactory functions. Therefore, combined study of molecular mechanisms underlying mood stabilization in olfactory tissue and tests of olfactory function could advance our understanding of the pathophysiology of BD and generate new data resource to study other pathoetiologies of BD. Furthermore, parallel studies of validated genes (from OE studies) in lymphoblasts (LB), confirmed by confirmed protein studies, may preempt future clinical and prognostic utility of blood markers. In specific aim 1 of this project, we propose to use our successful recruitment approach of BD subjects, to ascertain 40 non-medicated, non-smoking subjects with acute episodes of BD and 20 gender-, age-, and race- matched non-smoking normal controls (CTL) for this study. In specific aim 2, we will: a) compare BD and CTL on mRNA levels of (above-stated) 9 candidate genes of lithium action measured via RT-PCR, and b) examine the effect of lithium-associated changes in mRNA levels on clinical response (LR) in the BD subjects. To further illuminate cellular mechanisms of mood stabilization in specific aim 3, we will employ immunohistochemical techniques to examine densities of cell type-specific antibodies directed at the proliferating low-affinity nerve growth factor receptor (p75NGFR), postmitotic immature neurons (growth- associated protein 43 [GAP43]), and olfactory marker protein (OMP) in OE samples of all participants pre- and post-treatment, to determine if increased density of proliferating neuronal precursors and mature neurons (surrogate markers of neurogenesis and neuronal survival, respectively) are mediators of the effect of lithium- associated molecular alterations on LR. In other studies, we will: 1) examine differences in olfaction between BD and CTLs; 2) determine the association between baseline gene activity and olfactory function, and the impact of gender on these associations; 3) determine the utility of baseline olfactory function and lithium- associated changes in olfaction as surrogate markers of BD severity and mood stabilization; and 4) determine parallel dispositions between OE and LB on molecular markers of lithium action. Accomplishment of these aims will advance our understanding of the pathophysiology of BD, facilitate development of novel and improved therapeutics with specific desired effect on mood stabilization, and generate new data resource for team of scientists to further investigate other pathoetiologies of BD.

描述（由申请人提供）：双相情感障碍（BD）是一种具有重大社会负担的长期使人衰弱的疾病。我们对BD开发改善治疗疗法的能力的主要障碍是，我们对其病理生理学的理解是不完整的。锂，用于治疗BD的锂，改变了大量基因的稳态mRNA水平。值得注意的是GSK-32，肌醇单磷酸酶（Impase），MIP合酶，BCL-2，AKT，蛋白激酶C，MARCKS，胰岛素样生长因子结合蛋白-2（IGFBP-2）和ERK。但这仍然未知，如果这些分子变化中的任何一个会导致临床反应，如果有的话。迄今为止，研究活体BD患者中枢神经（CNS）组织中锂的细胞和分子作用是不可行的。尽管事后大脑对生物标志物的研究很有用，但它们不足以捕获依赖状态的BD分子特征。血细胞可能无法充分反映神经元细胞中的分子处置。在这项研究中，我们建议通过鼻腔活检使用嗅觉上皮（OE），并结合新型的激光捕获的显微减退，以阐明前瞻性临床对锂的临床反应的分子和细胞标记。之后，将确定这些分子模式在从同一受试者获得的淋巴细胞中的平行处置，以实现血液标记物的临床和预后效用。 OE是中枢神经系统的独特部分，它在一生中不断再生并区分为成熟的嗅觉受体神经元（ORN）。这是通过检查增殖（p75NGRF+）神经元前体的密度变化和成熟（嗅觉标记蛋白+）神经元（神经发生和OE神经元生存的替代标记）来检查BD神经发育假设的途径。同样特别感兴趣的是，与锂作用相关的基因失调，因此BD可能反映在嗅觉功能的缺陷中。因此，对嗅觉组织中情绪稳定和嗅觉功能测试的分子机制的结合研究可以提高我们对BD病理生理学的理解，并生成新的数据资源来研究BD的其他病因。此外，通过确认的蛋白质研究证实，经过验证的基因（来自OE研究）的平行研究（来自OE研究）可能会使未来的临床和预后效用。在该项目的特定目的1中，我们建议使用BD受试者的成功招募方法，以确定40个非药物，非吸烟的受试者，急性发作BD和20个性别，年龄和种族匹配的非 - 这项研究的吸烟正常对照（CTL）。在特定的目标2中，我们将：a）比较通过RT-PCR测量的锂作用的9个候选基因的BD和CTL，以及B）检查mRNA水平与锂相关变化对mRNA水平的影响BD受试者中的临床反应（LR）。为了进一步阐明特定目标3中情绪稳定的细胞机制，我们将采用免疫组织化学技术来检查针对增殖的低亲和力神经生长因子受体（P75NGFR）的细胞类型特异性抗体的密度43 [gap43]）和嗅觉标记蛋白（OMP）在所有参与者的OE样品中，以确定增殖神经元前体和成熟神经元的密度是否增加（分别锂相关分子改变对LR的影响的介体。在其他研究中，我们将：1）检查BD和CTL之间的嗅觉差异； 2）确定基线基因活性与嗅觉功能之间的关联，以及性别对这些关联的影响； 3）确定基线嗅觉功能和锂相关的嗅觉变化的效用，因为BD严重程度和情绪稳定的替代标志物； 4）确定在锂作用的分子标记上，确定OE和LB之间的平行处置。这些目标的完成将提高我们对BD的病理生理学的理解，促进新颖和改善的治疗学对情绪稳定的特定影响，并为科学家团队创造新的数据资源，以进一步研究BD的其他病因。