Organellar Na+/H+ Exchangers and Intracellular pH Regulation in Schizophrenia Brain

精神分裂症脑细胞器 Na /H 交换器和细胞内 pH 调节

• 批准号: 10448961
• 负责人: Brandon Scott Pruett
• 金额: $ 19.14万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10448961
• 关键词: AMPA Receptors; Acidity; Acute; Affect; Astrocytes; Autopsy; Biology; Brain; Calibration; Cell Culture Techniques; Cell Hypoxia; Cell membrane; Cells; Closure by clamp; Co-Immunoprecipitations; Confocal Microscopy; Development; Diagnosis; Disease; Disease model; Doctor of Philosophy; Endosomes; Energy Metabolism; Enrollment; Enzymes; Excision; Fluorescence; Funding; Future; Glycolysis; Goals; Health; Hypoxia; Individual; Knowledge; Label; Lead; Learning; Measures; Mediating; Mentorship; Metabolic; Methods; Molecular; Neurons; Neurotransmitter Receptor; Organelles; Patients; Peripheral; Pharmaceutical Preparations; Positioning Attribute; Post-Translational Protein Processing; Pre-Clinical Model; Prefrontal Cortex; Proteins; Psychiatry; Psychopathology; Regulation; Reporting; Research Personnel; Research Project Grants; Residencies; Resolution; Role; Scaffolding Protein; Schizophrenia; Sodium-Hydrogen Antiporter; Stress; Subcellular Fractions; Synapses; Techniques; Testing; Time; Tissues; Training; Transfection; Universities; Visualization; Work; base; brain tissue; cancer cell; career; cell type; cost; economic cost; experience; experimental study; first episode schizophrenia; high throughput screening; induced pluripotent stem cell; mortality; multimodality; neuroimaging; new therapeutic target; novel; novel therapeutics; professor; protein distribution; protein transport; ratiometric; receptor; response; severe mental illness; skills; stem cell differentiation; stressor; trafficking

Candidate: I am an Assistant Professor in UAB’s Department of Psychiatry with a background in molecular methods used in preclinical models and the role of intracellular pH dysregulation in neurodevelopmental illnesses. Additionally, I have expertise in diagnosis and treatment of psychopathology having received my MD- PhD from LSU Health Shreveport and completed a Psyhciatry Residency at Brown University. Career Goals and Development: I hope to gain expertise in assessing schizophrenia (SZ)-associated molecular disruptions in postmortem brain, in generating SZ patient-derived induced pluripotent stem cells (iPSCs) and differentiating them into disease relevant cell types, and in measuring cellular trafficking and luminal pH through the use of fluorescently-tagged protein constructs. By acquiring these skills and completing the studies laid out in this proposal, I will be well positioned and competitive for independent funding. Research Project: Deficits in protein post-translational modifications (PTM) and trafficking are reported in schizophrenia (SZ) brain, but the underlying cause is unknown. The function of organelles involved in PTM and trafficking is greatly impacted by pH disruptions, and Na+/H+ Exchangers (NHE) 6-9 are major regulators of organelle pH. In cancer cells, hypoxia causes altered energy metabolism and redistribution of NHE6 from endosomes to the plasma membrane. Similar metabolic alterations are reported in SZ brain suggesting that NHE6-9 intracellular distribution may also be affected, which could contribute to disrupted protein PTM and trafficking. So far, I have found that NHE7/8 expression is decreased in SZ dorsolateral prefrontal cortex (DLPFC) while NHE6/9 is unchanged. Still, NHE6/9 show increased expression in a tissue fraction enriched for synapses suggesting altered distribution of these proteins. Here, I propose to more extensively determine the expression and distribution of NHE6-9 first in SZ postmortem DLPFC and then in excitatory cortical neurons and astrocytes differentiated from patient-derived iPSCs. I will also determine how the introduction and removal of an acute stressor (hypoxia) affects the distribution of these proteins in these cells. Finally, I will transfect cells with fluorescently-tagged protein constructs to measure NHE6-9 and neurotransmitter receptor trafficking as well as organelle pH in live cells. These studies could help identify novel treatment targets for SZ and lead to high throughput assays to identify drugs that reverse SZ-associated molecular disruptions. Mentorship: The primary mentorship team for this proposal consists of Dr. James Meador-Woodruff, a world renowned expert in molecular disruptions in schizophrenia brain and analysis of postmortem brain tissue, Dr. Marek Napierala, an expert on molecular mechanisms of repeat expansion disorders and of modeling these illnesses using iPSCs differentiated into a variety of cell types including cortical neurons, and Dr. Vladimir Parpura, an expert in glial biology and visualization of vesicular trafficking in live cells through the use of fluorescently-tagged protein constructs.

应聘者：我是UAB精神病学系的助理教授，有分子方面的背景 临床前模型的方法和细胞内pH失调在神经发育中的作用 疾病。此外，在获得医学博士学位后，我在精神病理学方面拥有诊断和治疗方面的专业知识- 从路易斯安那州立大学健康中心获得博士学位，并在布朗大学完成精神病学住院医师学位。 职业目标和发展：我希望在评估精神分裂症(SZ)相关方面获得专业知识 在产生SZ患者来源的诱导多能干细胞过程中，死后脑中的分子破坏 (IPSCs)，并将其区分为与疾病相关的细胞类型，以及在测量细胞贩运和 通过使用荧光标记的蛋白质构建物来检测管腔pH值。通过获得这些技能并完成 在这项提案中列出的研究中，我将处于有利地位，并具有独立资金的竞争力。 研究项目：蛋白质翻译后修饰(PTM)缺陷和贩运在 精神分裂症(SZ)大脑，但潜在原因尚不清楚。细胞器在PTM中的作用和 PH值的变化对运输有很大的影响，Na+/H+交换器(NHE)6-9是主要的调节因子 细胞器pH值。在癌细胞中，低氧导致能量代谢改变和NHE6的重新分布 内涵体进入质膜。据报道，SZ大脑中也有类似的代谢变化，这表明 NHE6-9在细胞内的分布也可能受到影响，这可能导致蛋白PTM和 贩卖人口。到目前为止，我已经发现NHE7/8在深圳背外侧前额叶皮质的表达减少 (DLPFC)，而NHE6/9没有变化。尽管如此，NHE6/9在富含 突触表明这些蛋白质的分布发生了变化。在这里，我建议更广泛地确定 NHE6-9首先在死后DLPFC中的表达和分布，然后在兴奋性皮质神经元中的表达和分布 星形胶质细胞从患者来源的IPSCs分化而来。我还将决定如何介绍和 去除急性应激源(缺氧)会影响这些蛋白质在这些细胞中的分布。最后，我会 用荧光标记蛋白载体转染细胞检测NHE6-9和神经递质受体 活细胞的运输以及细胞器的pH值。这些研究可能有助于确定SZ的新治疗靶点 并导致高通量分析，以确定逆转SZ相关分子破坏的药物。 指导：这项提案的主要指导团队由詹姆斯·迈多-伍德拉夫博士组成，他是一个 在精神分裂症脑中的分子破坏和死后脑组织分析方面著名的专家。 Marek Napierala，研究重复扩张性疾病的分子机制并对其进行建模的专家 使用IPSCs的疾病分化为各种类型的细胞，包括皮质神经元和弗拉基米尔博士 神经胶质生物学和活细胞囊泡运输可视化的专家 荧光标记的蛋白质构建。