Function of the Zinc Finger Protein ZPR1 in neurodegeneration

锌指蛋白 ZPR1 在神经退行性变中的功能

• 批准号: 7887316
• 负责人: Laxman Dass Gangwani
• 金额: $ 32.16万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-15 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7887316
• 关键词: Ad5CMV; Address; Adenoviruses; Affect; American; Applications Grants; Axon; Biochemical; Cells; Clinical; Complex; Data; Defect; Development; Differentiation and Growth; Disease; Embryo; Exons; Family; Foundations; Genes; Genetic; Goals; Growth; Growth and Development function; Hereditary Disease; In Vitro; Individual; Infant Mortality; Infection; Investigation; Knockout Mice; Length; Longevity; Maintenance; Mediating; Molecular; Motor; Motor Neurons; Mus; Muscle; Mutation; Nerve Degeneration; Neuromuscular Diseases; Neurons; Nuclear; Nuclear Export; Nuclear Protein; Nuclear Proteins; Pathogenesis; Patients; Phenotype; Phosphorylation; Physiological; Play; Proteins; Research; Research Project Grants; Role; SMN protein (spinal muscular atrophy); SMN1 gene; SMN2 gene; Severities; Severity of illness; Siblings; Spinal; Spinal Cord; Spinal Muscular Atrophy; Testing; Tissues; Transgenic Mice; Transgenic Organisms; United States; Zinc Fingers; axon growth; axonal degeneration; cis trans isomerization; early childhood; in vivo; insight; mouse model; nerve supply; novel strategies; overexpression; plastin; postnatal; promoter; public health relevance; recombinase; snRNP Biogenesis; trafficking

DESCRIPTION (provided by applicant): The long-term goal of proposed research is to understand the physiological role of zinc finger protein ZPR1 in the pathogenesis of spinal muscular atrophy (SMA) caused by mutation of the survival motor neurons (SMN1) gene. SMA is characterized by degeneration of the spinal motor neurons. The cellular and molecular mechanisms of neuron degeneration are unclear and no treatment is available for SMA. The defect in nuclear accumulation of SMN is the biochemical defect in SMA. ZPR1 is required for nuclear accumulation of SMN in sub-nuclear bodies, including gems and Cajal bodies. The severity of SMA correlates negatively with SMN containing nuclear bodies. The severity of SMA may be influenced by the actions of modifier genes. One potential modifier gene is represented by ZPR1, which is down regulated in patients with SMA. The reduced expression of ZPR1 causes axonal defects and loss of spinal motor neurons in mice. The proposed research will determine the function of ZPR1 in the growth and differentiation of neurons and role of ZPR1 as a protective modifier of SMA. We will test the hypothesis that the increased expression of ZPR1 may ameliorate the severity of SMA disease and the ZPR1 gene may be a protective modifier of SMA. The Specific Aims of the research project are: Aim 1. To examine the physiological function of ZPR1 in neurons. We will examine the effect of ZPR1 deficiency in neurons (A) in vitro using cultured primary neurons, including spinal cord motor neurons from conditional Zpr1 (F1/F1) knockout mice and inactivation of the Zpr1 gene by infection with adenovirus expressing Cre recombinase. (B) In vivo by crossing conditional Zpr1 (F1/F1) mice with transgenic Hb9-cre mice expressing Cre recombinase under the control of endogenous mouse Hlxb9 promoter. Aim 2. To examine the effect of ZPR1 overexpression on the severity of disease and rescue of the SMA phenotype. (A) by generating a transgenic mouse overexpressing FLAG-ZPR1 under the control of a mouse ROSA26 promoter. We will examine the effect of ZPR1 overexpression on the growth and differentiation of neurons and growth and development of mice. (B) We will examine the effect of ZPR1 overexpression on severity and rescue of the SMA phenotype by crossing SMA model mice with transgenic Flag-Zpr1 mice. The phenotype of SMA mice with ZPR1 overexpression will be examined. Aim 3. To examine the role of ZPR1 in stabilization and nuclear accumulation of SMN. We will examine the cellular and molecular mechanisms of stabilization of SMN and nucleocytoplasmic trafficking of ZPR1 and SMN complexes. We will examine the role of cytoplasmic and nuclear proteins that interact with ZPR1 and may mediate nuclear export/import of ZPR1 and SMN. We will examine the role of phosphorylation and prolyl cis-trans isomerization that may regulate nucleocytoplasmic trafficking of ZPR1-SMN complexes. We will examine the effect of ZPR1 deficiency on the stability and integrity of SMN complex, nuclear accumulation of SMN and snRNP biogenesis. The findings of proposed study will provide insights into the function of ZPR1 in the nuclear accumulation of SMN and pathogenesis of SMA. PUBLIC HEALTH RELEVANCE: Spinal muscular atrophy (SMA), a neuromuscular disease caused by deficiency of the survival motor neuron (SMN) protein. SMA is one of the leading causes of infant mortality in the United States with an estimated 7.5 million Americans as carriers of disease. No treatment is available for this devastating genetic disease of early childhood. The severity of SMA disease is known to be influenced by the actions of modifier genes. The proposed study will determine the role of zinc finger protein ZPR1 in ameliorating the severity of SMA disease. Identification of ZPR1 as protective modifier of SMA will allow development of novel strategies for the treatment of SMA.

描述（由申请人提供）：拟议研究的长期目标是了解锌指蛋白ZPR 1在运动神经元存活（SMN 1）基因突变引起的脊髓性肌萎缩（SMA）发病机制中的生理作用。SMA的特征在于脊髓运动神经元的变性。神经元变性的细胞和分子机制尚不清楚，SMA尚无治疗方法。SMN的核积累缺陷是SMA的生化缺陷。ZPR 1是SMN在亚核体（包括宝石体和卡哈尔体）中积累所必需的。SMA的严重程度与含核小体的SMN呈负相关。SMA的严重程度可能受到修饰基因的影响。一个潜在的修饰基因是ZPR 1，它在SMA患者中下调。ZPR 1表达减少导致小鼠脊髓运动神经元轴突缺陷和缺失。这项研究将确定ZPR 1在神经元生长和分化中的功能以及ZPR 1作为SMA保护性修饰剂的作用。我们将检验ZPR 1表达增加可能改善SMA疾病的严重程度以及ZPR 1基因可能是SMA的保护性修饰剂的假设。研究项目的具体目标是：目标1。研究ZPR 1在神经元中的生理功能。我们将使用培养的原代神经元（包括来自条件性Zpr 1（F1/F1）敲除小鼠的脊髓运动神经元）和通过用表达Cre重组酶的腺病毒感染的Zpr 1基因的失活，在体外检查ZPR 1缺陷在神经元中的作用（A）。(B)通过将条件性Zpr 1（F1/F1）小鼠与在内源性小鼠Hlxb 9启动子控制下表达Cre重组酶的转基因Hb 9-cre小鼠进行体内杂交。目标2.研究ZPR 1过表达对疾病严重程度和SMA表型挽救的影响。(A)通过产生在小鼠ROSA 26启动子控制下过表达FLAG-ZPR 1的转基因小鼠。我们将研究ZPR 1过表达对神经元生长和分化以及小鼠生长发育的影响。(B)我们将通过将SMA模型小鼠与转基因Flag-Zpr 1小鼠杂交来检查ZPR 1过表达对SMA表型的严重程度和挽救的影响。将检查具有ZPR 1过表达的SMA小鼠的表型。目标3.研究ZPR 1在SMN稳定和核积累中的作用。我们将研究SMN的稳定和ZPR 1和SMN复合物的核质运输的细胞和分子机制。我们将研究与ZPR 1相互作用的细胞质和核蛋白的作用，并可能介导ZPR 1和SMN的核输出/输入。我们将研究磷酸化和脯氨酰顺反异构化的作用，可能会调节ZPR 1-SMN复合物的核质运输。我们将研究ZPR 1缺陷对SMN复合体的稳定性和完整性、SMN的核积累和snRNP生物发生的影响。本研究的结果将有助于深入了解ZPR 1在SMN核积聚和SMA发病机制中的作用。 公共卫生相关性：脊髓性肌萎缩症（SMA），一种由运动神经元存活（SMN）蛋白缺乏引起的神经肌肉疾病。SMA是美国婴儿死亡的主要原因之一，估计有750万美国人是疾病携带者。对于这种幼儿期的毁灭性遗传疾病，目前尚无治疗方法。SMA疾病的严重程度已知受修饰基因作用的影响。这项拟议的研究将确定锌指蛋白ZPR 1在改善SMA疾病严重程度方面的作用。鉴定ZPR 1作为SMA的保护性修饰剂将允许开发SMA治疗的新策略。