Nanoelectrode arrays for study of the molecular mechanisms, triggers, and inhibit

用于研究分子机制、触发和抑制的纳米电极阵列

基本信息

批准号：
7778178
负责人：
PETER J. BURKE
金额：
$ 24.32万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-03-15 至 2013-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7778178
关键词：
Action Potentials Address Adenine Nucleotides Apoptosis Apoptosis Inhibitor Apoptosis Regulator Apoptotic Binding Biochemical Caliber Cancer Biology Carbon Nanotubes Cell Death Cells Charge Cholesterol Cytosol Defect Diamide Down-Regulation Electrodes Functional disorder Generations Genes Glucose Heme Hepatocyte Hexokinase 2 Individual Induction of Apoptosis Infusion procedures Inner mitochondrial membrane Ions Kinetics Lipids Maintenance Measures Membrane Membrane Potentials Metabolism Metals Microfluidics Mitochondria Mitochondrial DNA Mitochondrial Proteins Mitochondrial Proton-Translocating ATPases Mitochondrial Swelling Molecular Monitor Mutation Neurons Oxidative Stress Oxygen Peroxides Phase Phospholipids Potential Energy Primary carcinoma of the liver cells Process Production Protein Family Protein Isoforms Regulation Repression Resolution Signal Transduction Signal Transduction Pathway Source Surface Swelling System Technology Testing Variant Vesicle aerobic glycolysis cancer cell cell killing complex IV coproporphyrinogen III cytochrome c oxidase falls functional status high throughput technology inhibitor/antagonist macromolecule miniaturize mitochondrial dysfunction mitochondrial membrane mitochondrial permeability transition pore nano nanowire oligomycin sensitivity-conferring protein response seal sensor single walled carbon nanotube theories tumorigenesis uptake

项目摘要

DESCRIPTION (provided by applicant): Mitochondria are the central regulator of apoptosis, a process initiated by the activation of the mitochondrial permeability transition pore (mtPTP), an aggregate of several mitochondrial proteins. When this pore opens, the critical membrane polarization of the mitochondrial inner membrane disappears and ions equilibrate between the matrix and cytosol resulting in mitochondrial swelling. This leads to release of the contents of the mitochondrial intermembrane space into the cell cytosol, including a number of cell death promoting factors killing the cell. The mtPTP can be activated by uptake of excessive Ca++; increased oxidative stress; decreased mitochondrial membrane potential, and reduced ADP and ATP. It is generally agreed upon that repression of apoptosis is one of the fundamental steps in tumorigenesis. Cancer cells acquire unresponsiveness to apoptosis facilitating signals, thus enabling uncontrolled proliferation. For this reason, the induction of apoptosis is one of the modes of actions of chemotherapeutic compounds. In order to allow further high throughput studies of the biochemical facilitators and inhibitors, of apoptosis, and to determine if changes in individual mitochondrial membrane potential P are important to cellular metabolism, we need to develop a system to monitor P in individual mitochondria. To accomplish this objective, we propose to extend studies that have monitored the action potentials in neurons using an array of parallel electrodes to which the mitochondria are adhered. Our thesis is that a nanoelectrode technology can be developed to capacitively measure membrane potential across the mitochondrial inner membrane phospholipid bilayer without actually penetrating the membrane. We propose to develop nano-electrical transduction sensor arrays with sufficiently high spatial and temporal resolution to monitor the charge changes on the surface of a mitochondrion sized lipid vesicle and the individual mitoplast. With this technology, we will then interrogate the regulation of P in normal and cancer cells. Several key features on mitochondrial metabolism are now recognized as important to the alteration of cancer cell mitochondrial function: changes in the Akt signal transduction pathway, induction of hexokinase II, alteration an adenine nucleotide translocator (ANT) isoform expression, down regulation of the SOC2 cytochrome c oxidase (complex IV, COX) assembly factor, mutation in mitochondrial DNA (mtDNA) genes, and modulation of the mitochondrial permeability transition pore (mtPTP) and its interaction with the pro- and anti- apoptotic Bcl2 family proteins. While all of these are important factors in the alteration of cancer cell metabolism, they still fall short of explaining the near universal alterations in mitochondrial function observed in cancer cells. A high throughput technology to monitor P in mitochondria will allow further studies of these issues in cancer biology.

描述（由申请人提供）：线粒体是细胞凋亡的中心调节剂，这是通过激活线粒体通透性过渡孔（MTPTP）引发的，这是几种线粒体蛋白的聚集体。当这种孔打开时，线粒体内膜的临界膜极化消失，离子在基质和胞质溶胶之间平衡，导致线粒体肿胀。这导致线粒体膜间空间的含量释放到细胞胞质中，包括许多细胞死亡促进因子杀死细胞。 MTPTP可以通过吸收过多的Ca ++激活；增加氧化应激；线粒体膜电位降低，ADP和ATP降低。人们普遍认为，抑制凋亡是肿瘤发生的基本步骤之一。癌细胞对凋亡促进信号的无反应性，从而导致不受控制的增殖。因此，凋亡的诱导是化学治疗化合物作用的一种模式之一。为了允许对生化促进剂和抑制剂，凋亡的进一步高通量研究，并确定单个线粒体膜电位P的变化是否对细胞代谢很重要，我们需要开发一个系统来监测单个线粒体中P的P。为了实现这一目标，我们建议使用一系列平行电极来扩展对神经元的作用电位的研究，并遵守线粒体。我们的论点是，可以开发出一种纳米电极技术来测量线粒体内膜磷脂双层的膜电位，而无需实际穿透膜。我们建议开发具有足够高的空间和时间分辨率的纳米电气转导传感器阵列，以监视线粒体大小的脂质囊泡表面的电荷变化和单个线粒体。利用这项技术，我们将在正常细胞和癌细胞中询问P的调节。 Several key features on mitochondrial metabolism are now recognized as important to the alteration of cancer cell mitochondrial function: changes in the Akt signal transduction pathway, induction of hexokinase II, alteration an adenine nucleotide translocator (ANT) isoform expression, down regulation of the SOC2 cytochrome c oxidase (complex IV, COX) assembly factor, mutation in mitochondrial DNA （mtDNA）基因，以及线粒体通透性过渡孔（MTPTP）的调节及其与促凋亡BCL2家族蛋白的相互作用。尽管所有这些都是癌细胞代谢改变的重要因素，但它们仍然没有解释癌细胞中观察到的线粒体功能的几乎普遍变化。高通量技术以监测线粒体的P，将允许对癌症生物学中的这些问题进行进一步研究。