Mediators of Lithium Action in Olfactory Epithelium

嗅觉上皮细胞中锂作用的介质

• 批准号: 8189220
• 负责人: Evaristus A Nwulia
• 金额: $ 58.2万
• 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/8189220
• 关键词: Acute; Adult; Aftercare; Age; Antibodies; Autopsy; 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; Epithelial; 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. PUBLIC HEALTH RELEVANCE: We are well overdue for better understanding of the pathophysiology of, and for development of improved therapeutics of Bipolar disorder (BD), a condition that causes extensive illness burden to sufferers and substantial economic burden to the society. We have now developed novel techniques to prospectively study cellular and molecular determinants of mood stabilization during lithium treatment, using a central nervous system tissue, olfactory epithelium (OE) of acutely ill BD patients. The goals of this proposal are: 1) recruit autely ill BD patients to assess clinical symptoms and obtain repeated OE biopsies, before and after lithium monotherapy; 2) identify molecular markers whose alterations predict clinical response; 3) determine if the effect of lithium-induced molecular alterations is mediated through enhanced neurogenesis and neuronal survival; and 4) examine olfactory function and gene activity in blood cells for utility as markers of disease-associated, lithium-modifiable molecular dysregulation of BD. This study could enhance development of targeted therapeutics for BD.

描述（由申请人提供）：双相情感障碍（BD）是一种慢性衰弱性疾病，具有巨大的社会负担。我们开发改善双相情感障碍疗法的能力的一个主要障碍是我们对其病理生理学的理解不完整。用于治疗 BD 的锂会改变大量基因的稳态 mRNA 水平；特别是 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。但目前尚不清楚这些分子变化如何带来临床反应（如果有的话）。迄今为止，研究锂对活体 BD 患者中枢神经 (CNS) 组织的细胞和分子影响尚不可行。尽管死后大脑对于生物标志物的研究很有用，但它们不足以捕获 BD 的状态依赖性分子特征。血细胞可能无法充分反映神经元细胞中的分子分布。在这项研究中，我们建议通过鼻活检使用嗅上皮（OE），结合新型激光捕获显微切割来阐明前瞻性确定的锂临床反应的分子和细胞标志物。随后将确定从同一受试者获得的淋巴母细胞中这些分子模式的平行分布，以用于血液标志物的可能的临床和预后用途。 OE 是 CNS 的独特部分，在整个生命过程中不断再生并分化为成熟的嗅觉受体神经元 (ORN)。这为通过检查增殖（p75NGRF+）神经元前体和成熟（嗅觉标记蛋白+）神经元（OE 中神经发生和神经元存活的替代标记）密度的变化来检查 BD 的神经发育假说提供了一条途径。同样特别令人感兴趣的是，与锂作用相关的基因失调，因此 BD 可能反映在嗅觉功能缺陷中。因此，结合研究嗅觉组织情绪稳定的分子机制和嗅觉功能测试可以增进我们对BD病理生理学的理解，并为研究BD的其他病理学产生新的数据资源。此外，对淋巴母细胞 (LB) 中经过验证的基因（来自 OE 研究）的平行研究，经已证实的蛋白质研究证实，可能会抢占血液标记物未来的临床和预后用途。在该项目的具体目标 1 中，我们建议利用我们成功招募 BD 受试者的方法，确定 40 名患有 BD 急性发作且未接受药物治疗、不吸烟的受试者，以及 20 名性别、年龄和种族匹配的非本研究的吸烟正常对照（CTL）。在具体目标 2 中，我们将：a) 比较 BD 和 CTL 对通过 RT-PCR 测量的（上述）9 个锂作用候选基因的 mRNA 水平，以及 b) 检查锂相关的 mRNA 水平变化对BD 受试者的临床反应（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）、有丝分裂后未成熟神经元（生长相关蛋白）的细胞类型特异性抗体的密度。 43 [GAP43]），以及所有参与者治疗前和治疗后的 OE 样本中的嗅觉标记蛋白（OMP），以确定增殖密度是否增加神经元前体和成熟神经元（分别是神经发生和神经元存活的替代标记）是锂相关分子改变对 LR 影响的介质。在其他研究中，我们将：1）检查 BD 和 CTL 之间嗅觉的差异； 2）确定基线基因活性和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嗅觉变化作为 BD 严重程度和情绪稳定性替代标记的效用； 4) 确定 OE 和 LB 在锂作用分子标记上的平行配置。这些目标的实现将增进我们对 BD 病理生理学的理解，促进对情绪稳定具有特定预期效果的新型和改进疗法的开发，并为科学家团队进一步研究 BD 的其他病理学产生新的数据资源。 公共卫生相关性：我们早就该更好地了解双相情感障碍 (BD) 的病理生理学并开发改进的治疗方法，双相情感障碍 (BD) 是一种给患者带来广泛疾病负担并给社会带来巨大经济负担的疾病。我们现在已经开发出新技术，利用急性双相情感障碍患者的中枢神经系统组织、嗅觉上皮 (OE)，前瞻性研究锂治疗期间情绪稳定的细胞和分子决定因素。该提案的目标是：1）招募患有严重双相情感障碍的患者，以评估临床症状并在锂单一疗法之前和之后进行重复的 OE 活检； 2) 识别分子标志物，其改变可预测临床反应； 3) 确定锂诱导的分子改变的影响是否是通过增强神经发生和神经元存活来介导的； 4) 检查血细胞中的嗅觉功能和基因活性，以用作 BD 疾病相关的锂可修饰分子失调的标记。这项研究可以促进双相情感障碍靶向治疗的开发。