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Mechanisms of Neuronal Death and Neuroprotection

神经元死亡和神经保护机制

基本信息

批准号：
7743995
负责人：
ANTHONY John WINDEBANK
金额：
$ 29万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-06-01 至 2011-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7743995
关键词：
Acute Adenovirus Vector Adverse effects Animal Model Apoptosis Binding Biological Assay Carboplatin Catalytic Domain Cell Cycle Cell Death Cessation of life Chronic Cisplatin Clinical Cyclin D1 Cysteine DNA DNA Adducts DNA Binding DNA Damage DNA Synthesis Inhibitors DNA biosynthesis DNA lesion Dose Dose-Limiting Exhibits Exposure to Genetic Transcription Glutamate-Cysteine Ligase Glutathione Goals Growth Factor Hour In Vitro Knockout Mice Lead Ligase Measures Mediating Methods Mitochondria Mitochondrial DNA Mus Nerve Growth Factors Neuronal Injury Neurons Neuropathy Nuclear Pathway interactions Pharmaceutical Preparations Pigment Epithelium Platinum Platinum Compounds Platinum adduct Polymerase Chain Reaction R-DNA RNA chemical synthesis Research Personnel Respiratory Chain Reverse Transcriptase Polymerase Chain Reaction Rodent Spinal Ganglia Testing Therapeutic Time Viral Viral Vector Zalcitabine base cancer therapy clinically relevant drug withdrawal glutathione synthase in vivo mitochondrial genome neuron loss neuroprotection neurotoxic novel novel strategies oxaliplatin prevent programs repaired sensory neuropathy

项目摘要

The use of platinum compounds is increasing in the treatment of cancer. Cisplatin, carboplatin and oxaliplatin produce a dose-related and dose-limiting sensory neuropathy. We have demonstrated that cisplatin binds to neuronal DNA, activates DMA-damage recognition pathways, initiates aberrant cell cycle entry, and induces apoptosis in vitro and in vivo. We now propose to test the hypothesis that platinum compounds produce neuronal injury by a common mechanism that involves separate nuclear (n-DNA) and mitochondrial DNA (mt-DNA) binding followed by synergistic activation of parallel death pathways. A new approach to measuring mt-DNA platination has been developed using inhibition of the polymerase chain reaction. We will determine whether the number of platinum adducts in mt-DNA of DRG neurons is sufficient to prevent replication and transcription of the mitochondrial genome. Function of respiratory chain components will be measured. Inability to repair Pt-DNA lesions in mt-DNA would result in attrition of mitochondria and chronic neuronal death explaining the clinical phenomenon of "coasting" or progression of neuropathy after drug cessation. We will use DRG neurons from Bax and cyclin D1 knockout mice to determine whether platinum-induced inhibition of mitochondrial function is sufficient to cause neuronal death. We will use the mitochondrial DNA synthesis inhibitor dideoxycytidine (ddC) to determine whether inhibition of mitochondrial DNA replication is independently sufficient to cause cell death. The mitochondrial genome will be protected by selectively increasing mitochondrial glutathione. The modifier and catalytic subunits ofglutamate cysteine ligase (GCL) andglutathione synthetase (GS) willbe targeted to mitochondria in an adeno-viral vector to reduce formation of mt-DNA adducts. Both nerve growth factor (NGF) and pigment epithelium derived growth factor (PEDF) have been demonstrated to partially protect DRG from cisplatin-induced apoptosis. We will determine whether a combination of therapeutic strategies that block n-DNA induced apoptosis and protect mt-DNA promote long-term survival of cisplatin treated DRG in vitro. If the combination strategy is effective,we will test it in an animal model by developing methods to provide long-term delivery of growth factors and viral targeting of GCL and GS to DRG in vivo. The goal is to develop a mechanism-based therapy that will prevent the major dose-limiting side effect of the platinum compounds.

铂化合物在癌症治疗中的使用正在增加。顺铂、卡铂和奥沙利铂产生剂量相关和剂量限制性感觉神经病变。我们已经证明顺铂与神经元DNA结合，激活DMA损伤识别途径，启动异常细胞周期进入，并在体外和体内诱导凋亡。我们现在提出检验铂化合物通过一种共同的机制产生神经元损伤，该机制涉及分离的核（n-DNA）和线粒体DNA（mt-DNA）结合，随后协同激活平行死亡途径。一个新利用聚合酶链的抑制作用，发展了一种测量线粒体DNA铂化的方法反应我们将确定背根神经节神经元线粒体DNA中铂加合物的数量是否足够以阻止线粒体基因组的复制和转录。呼吸链功能将对组件进行测量。不能修复线粒体DNA中的Pt-DNA损伤将导致线粒体DNA的磨损。线粒体和慢性神经元死亡解释了“滑行”或进展的临床现象，停药后神经病变。我们将使用Bax和细胞周期蛋白D1敲除小鼠的DRG神经元，确定铂诱导的线粒体功能抑制是否足以导致神经元死亡。我们将使用线粒体DNA合成抑制剂双脱氧胞苷（ddC）来确定是否抑制线粒体DNA复制的独立性足以导致细胞死亡。线粒体基因组将通过选择性增加线粒体谷胱甘肽来保护。的谷氨酸半胱氨酸连接酶（GCL）和谷胱甘肽合成酶（GS）的修饰和催化亚基将被靶向腺病毒载体中的线粒体以减少mt-DNA加合物的形成。神经生长已经证明，神经生长因子（NGF）和色素上皮衍生生长因子（PEDF）部分保护DRG免受顺铂诱导的凋亡。我们将确定是否联合治疗阻断n-DNA诱导的细胞凋亡和保护线粒体DNA的策略促进顺铂的长期存活体外处理DRG。如果组合策略有效，我们将通过开发提供生长因子的长期递送以及GCL和GS体内病毒靶向DRG的方法。目的是开发一种基于机制的治疗方法，以防止主要的剂量限制性副作用，铂化合物。

项目成果

期刊论文数量（15）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cisplatin induced mitochondrial DNA damage in dorsal root ganglion neurons.

DOI：
10.1016/j.nbd.2010.11.017
发表时间：
2011-03
期刊：
NEUROBIOLOGY OF DISEASE
影响因子：
6.1
作者：
Podratz, Jewel L.;Knight, Andrew M.;Ta, Lauren E.;Staff, Nathan P.;Gass, Jennifer M.;Genelin, Konstantin;Schlattau, Alexander;Lathroum, Liselle;Windebank, Anthony J.
通讯作者：
Windebank, Anthony J.

Controlled release of NFkappaB decoy oligonucleotides from biodegradable polymer microparticles.

从可生物降解的聚合物微粒中控制释放 NFkappaB 诱饵寡核苷酸。

DOI：
10.1016/s0142-9612(01)00409-4
发表时间：
2002
期刊：
Biomaterials
影响因子：
14
作者：
Zhu,Xun;Lu,Lichun;Currier,BradfordL;Windebank,AnthonyJ;Yaszemski,MichaelJ
通讯作者：
Yaszemski,MichaelJ

Bortezomib alters microtubule polymerization and axonal transport in rat dorsal root ganglion neurons.

DOI：
10.1016/j.neuro.2013.09.001
发表时间：
2013-12
期刊：
NEUROTOXICOLOGY
影响因子：
3.4
作者：
Staff, Nathan P.;Podratz, Jewel L.;Grassner, Lukas;Bader, Miranda;Paz, Justin;Knight, Andrew M.;Loprinzi, Charles L.;Trushina, Eugenia;Windebank, Anthony J.
通讯作者：
Windebank, Anthony J.

Methylmercury induces apoptosis in cultured rat dorsal root ganglion neurons.

甲基汞诱导培养的大鼠背根神经节神经元细胞凋亡。