Application of plant genomics to alcoholic brain damage

植物基因组学在酒精性脑损伤中的应用

• 批准号: 6950159
• 负责人: JOHN M. LITTLETON
• 金额: $ 5.43万
• 依托单位: NAPROGENIX, INC
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-15 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6950159
• 关键词: Agrobacterium tumefaciens; alcoholism /alcohol abuse; antioxidants; biotechnology; biotherapeutic agent; brain injury; drug screening /evaluation; ethanol; fluorescence microscopy; gas chromatography mass spectrometry; gene expression; genetically modified plants; high throughput technology; molecular cloning; molecular probes; neural degeneration; neuropharmacology; neuroprotectants; neurotoxicology; oxidation; oxidative stress; plant extracts; plant genetics; protoplast /spheroplast; tissue /cell culture

DESCRIPTION (provided by applicant): Alcoholic liver damage (ALD) is among the ten most common causes of death in the USA. The liver may be uniquely sensitive because it metabolizes ethanol at a high rate. This generates acetaldehyde intracellularly, damaging cell components directly, and/or inducing apoptotic cell death. Although some plant natural products are protective it would be of value to discover agents with greater efficacy against ALD. Here, a novel biotechnology, based on selection of mutant plant cells for resistance to alcohol, followed by high throughput pharmacological screening (HTPS) of extracts from surviving cells, is applied to this problem. The mutant population will be generated by activation tagging mutagensis, and extracts from the resulting alcohol-resistant clonal cultures screened for protection of hepatocyte cultures against ethanol-induced damage. However, because plant cells in culture metabolize ethanol to a very limited extent, we propose first to produce transgenic plant lines (N tabacum) over-expressing the activated gene for plant or mammalian ADH. Cell cultures derived from these plants will be evaluated for similarities to mammalian liver cells in metabolism of ethanol to acetaldehyde. Phase 1 is the creation of these ADH-expressing (ADHE) lines. In subsequent phases, hepatoprotective compounds from resitant clonal extracts will be tentatively identified by GC/MS. Regardless of whether novel compounds are discovered, the ADHE plant lines, and the ethanol resistant clonal cultures are patentable and represent a valuable resource for development of alcohol-related medications in the future.

描述（由申请人提供）：酒精性肝损伤 (ALD) 是美国十大最常见的死亡原因之一。肝脏可能特别敏感，因为它以很高的速度代谢乙醇。这会在细胞内产生乙醛，直接损害细胞成分，和/或诱导细胞凋亡。尽管一些植物天然产物具有保护作用，但发现对 ALD 更有效的药物将很有价值。在这里，一种新颖的生物技术应用于解决这个问题，该技术基于选择耐酒精的突变植物细胞，然后对存活细胞的提取物进行高通量药理学筛选（HTPS）。突变体群体将通过激活标记诱变产生，并筛选所得的耐酒精克隆培养物的提取物，以保护肝细胞培养物免受乙醇诱导的损伤。然而，由于培养中的植物细胞代谢乙醇的程度非常有限，我们建议首先生产过度表达植物或哺乳动物ADH激活基因的转基因植物系（N tabacum）。将评估源自这些植物的细胞培养物与哺乳动物肝细胞在乙醇代谢为乙醛方面的相似性。第一阶段是创建这些表达 ADH (ADHE) 的细胞系。在后续阶段，将通过 GC/MS 初步鉴定耐药克隆提取物中的保肝化合物。 无论是否发现新化合物，ADHE 植物系和乙醇抗性克隆培养物都可以获得专利，并且代表了未来酒精相关药物开发的宝贵资源。