Gene Therapy And Neurodegeneration

基因治疗和神经退行性疾病

• 批准号: 8553237
• 负责人: Brandon Harvey
• 金额: $ 27.33万
• 依托单位: NATIONAL INSTITUTE ON DRUG ABUSE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8553237
• 关键词: Appearance; Axon; Behavioral; Brain; Brain-Derived Neurotrophic Factor; Caenorhabditis elegans; Cell Culture Techniques; Cell Transplantation; Cells; Cellular Stress; Chimeric Proteins; Collaborations; Dendrites; Disease; Drug toxicity; Event; Extracellular Matrix Proteins; GDNF gene; Gene Delivery; Gene Proteins; Genes; Glutamate Transporter; Glutamates; Goals; Green Fluorescent Proteins; Homologous Gene; Illicit Drugs; Ischemia; Ischemic Brain Injury; Laboratories; Lead; Link; Manuscripts; Methamphetamine; Modeling; National Institute of Drug Abuse; Nerve Degeneration; Neurons; Parkinson Disease; Protein Region; Proteins; Publications; Publishing; Reagent; Recovery; Rodent; Rodent Model; Role; Serotyping; Stroke; Structure-Activity Relationship; Testing; Therapeutic; Tissues; Toxic effect; Toxin; Transgenes; Work; adeno-associated viral vector; bone morphogenetic protein 7; cell type; dopaminergic neuron; endoplasmic reticulum stress; gene therapy; in vitro Model; interest; mouse model; neuroprotection; neurotoxicity; neurotrophic factor; novel; stable cell line; therapeutic gene; tool; vector

Over the past several years, our group has been focused on studying genes with neuromodulatory, neuroprotective and/or neuroregenerative effects in models of neurodegeneration and neurotoxicity. As part of a collaboration on a primary project of Dr. Barry Hoffer (NIDA) and Mart Saarma (U Helsinki), we have examined the neuroprotective effects of conserved dopaminergic neurotrophic factor (CDNF) in an mouse model of Parkinsons disease. We have found that CDNF protein can confer neuroprotection and neuroregeneration against neurotoxicity caused by the dopaminergic neuron toxin, MPTP. The manuscript describing this study has been accepted for publication at Cell Transplantation. We are also working on the homolog to CDNF, mesencephalic astrocyted derived neurotrophic factor (MANF). We published a study last year describing the neuroprotective actions of MANF against ischemic brain injury. We are continuing to explore the neuroprotective and neuroregenerative mechanism(s) MANF/CDNF. Towards this goal, we are preparing a manuscript describing the function of MANF in C. elegans as it relates to cellular stress, specifically, endoplasmic reticulum (ER) stress. The phenomenon of ER stress occurs in many diseases beyond neurodegenerative and understanding its role may lead to broader therapeutic strategies for MANF and CDNF. In addition to our work with C. elegans, we have established several novel reagents for studying the structure/function relationship of MANF in ER stress. Using stable cell lines expressing a fusion proteins of MANF and green fluorescent protein (GFP), we have identified a region of the protein important for its localization/secretion. Using the tools weve built over the past year, we will focus on understanding MANFs function in the cell. Our section has previously demonstrated the ability of Bone Morphogenetic Protein 7 (BMP7) to promote neuroregeneration. As a continuation of this work, we have used an in vitro model of primary cortical neurons to show that BMP7 changes the appearance of axons and dendrites and have linked this to the change in extracellular matrix proteins. We are preparing the manuscript for publication. In addition to neurotrophic factors, we have completed a study examining the ability of the glutamate transporter (GLT1) to reduce the toxic effects of glutamate using an rodent model of stroke. We generated an adeno-associated vector expressing the GLT1 gene and demonstrated it could increase glutamate clearance in the brain caused by an ischemic event. This correlated with increased behavioral recovery and decreased tissue damage following ischemia. The results were published in PLoSONE and emphasize the importance of targeting ischemia-induced glutamate overflow acutely following stroke. ct is ongoing.

在过去的几年中，我们的小组一直专注于研究神经性变性和神经变性和神经毒性模型中具有神经调节，神经保护性和/或神经产生效应的基因。 作为Barry Hoffer博士（NIDA）和Mart Saarma（U Helsinki）的主要项目的一部分，我们研究了保守多巴胺能神经营养因子（CDNF）的神经保护作用。 我们发现，CDNF蛋白可以赋予由多巴胺能神经元毒素MPTP引起的神经毒性的神经保护和神经创生。 描述这项研究的手稿已被接受在细胞移植时出版。 我们还在研究中脑体形胶质细胞衍生的神经营养因子（MANF）的CDNF同源物。 去年，我们发表了一项研究，描述了MANF对缺血性脑损伤的神经保护作用。 我们正在继续探索神经保护性和神经创造机制。 为了实现这一目标，我们正在准备一个手稿，描述了秀丽隐杆线虫在秀丽隐杆线虫中的功能，因为它与细胞应激有关，具体是内质网（ER）应激。 ER应力的现象发生在神经退行性以外的许多疾病中，了解其作用可能会导致对MANF和CDNF的更广泛的治疗策略。 除了与秀丽隐杆线虫的合作外，我们还建立了几种新型试剂，用于研究MANF在ER应力下的结构/功能关系。 使用表达MANF和绿色荧光蛋白（GFP）融合蛋白（GFP）的稳定细胞系，我们已经确定了该蛋白的局部定位/分泌重要区域。 使用过去一年中我们构建的工具，我们将专注于了解单元格中的MANFS功能。 我们的部分先前已经证明了骨形态发生蛋白7（BMP7）促进神经变成的能力。 作为这项工作的延续，我们使用了原代皮质神经元的体外模型，以表明BMP7改变了轴突和树突的外观，并将其与细胞外基质蛋白的变化联系起来。 我们正在准备手稿以供出版。 除了神经营养因素外，我们还完成了一项研究，研究了谷氨酸转运蛋白（GLT1）使用中风模型降低谷氨酸毒性作用的能力。 我们产生了表达GLT1基因的腺相关载体，并证明它可能会增加由缺血性事件引起的大脑中的谷氨酸清除率。 这与缺血后的行为恢复增加和组织损伤减少有关。 该结果在Plosone中发表，并强调靶向缺血诱导的谷氨酸溢流在中风后的重要性。 CT正在进行中。