LENS EPITHELIUM AND HELIUM ION CATARACTOGENESIS

晶状体上皮和氦离子白内障发生

• 批准号: 2711124
• 负责人: ELEANOR A BLAKELY
• 金额: $ 29.62万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2711124
• 关键词: DNA damage; HeLa cells; SDS polyacrylamide gel electrophoresis; cataract; cell differentiation; cellular pathology; chromosome aberrations; cytogenetics; enzyme linked immunosorbent assay; eye neoplasms; fibroblast growth factor; helium; human tissue; iatrogenic disease; in situ hybridization; lens proteins; melanoma; messenger RNA; neoplasm /cancer radiation therapy; northern blottings; radiation genetics; radiation therapy dosage; tissue /cell culture; tissue mosaicism; western blottings

Cataracts arise in uveal melanoma patients as a complication following their successful treatment with helium-ion radiotherapy. The objective of the proposed research is to determine the helium-ion-induced alterations in chromosomes and in protein expression that are important to cataractogenesis, and to develop strategies to diminish the incidence or severity of these changes. The current paradigm for cataractogenesis is focussed on genomic damage of lens epithelial cells leading to altered crystallin proteins. This proposal will examine two alternative mechanisms of cataractogenesis involving radiation-induction of basic Fibroblast Growth Factor (bFGF) in human lens epithelial (HLE) cells functioning either to alter the normal program of crystallin expression and thereby disrupting normal fiber formation, or the radiation-induced bFGF acting to hinder cell loss processes which leads to the formation of aberrant lens fiber formation. To this end experiments have been designed with three specific aims that will: Firstly, characterize acute and residual responses to helium-ion irradiation of cultured HLE cells grown on extracellular matrix. Quantitative dose-response measurements between helium-ion induced survival, repair capacity and yields of micronuclei, apoptotic nuclei, and persistent chromosomal rearrangements will be determined. Secondly, since radiation is known to induce bFGF in endothelial cells leading to inhibition of apoptosis, experiments are proposed that will determine whether helium-ion radiation changes levels of bFGF mRNA or protein in HLE cells, and whether there is a correlation with apoptotic events. Finally, an investigation of the possible modification of intracellular lens proteins by helium-ion radiation is proposed. The program will elucidate relationships between helium-ion- induced damage to the chromatin of HLE cells in vitro, and biological consequences to the cells surviving the resulting damage. In the revised application significant changes include: 1) an increased source of HLE cells, 2) new preliminary data from mRNA and protein analyses (SDS-PAGE) of exponential and confluent cell cultures, 3) new preliminary data with immunological evidence confirming feasibility to characterize lens cell differentiation, 4) some revised experimental protocols, and 5) and an updated review of literature published in this rapidly increasing area. This knowledge will allow correlative comparisons with available experimental work in vivo, and may provide a basis for potential changes to the treatment protocol that could reduce the risk of cataract. The biological mechanisms contributing to cataractogenesis may also be elucidated.

葡萄膜黑色素瘤患者出现白内障是以下的并发症 他们成功地接受了氦离子放射治疗。目标是 建议的研究是确定氦离子诱导的 重要的染色体和蛋白质表达的变化 白内障的发生，并制定减少发病率的策略 或这些变化的严重性。当前白内障发生的研究范式 集中在晶状体上皮细胞的基因组损伤导致的改变 晶体蛋白。这项提案将研究两种替代方案 BASIC辐射诱发白内障的发生机制 成纤维细胞生长因子对人晶状体上皮细胞的影响 作用是改变晶体蛋白表达的正常程序 从而扰乱正常的纤维形成，或辐射诱导的 碱性成纤维细胞生长因子阻止细胞丢失过程，从而导致细胞的形成 晶状体纤维的异常形成。为此，已经进行了实验 设计有三个具体目标：第一，刻画急性 培养的HLE细胞对氦离子照射的残余反应 生长在细胞外基质上。定量剂量-反应测量 氦离子诱导的细胞存活率、修复能力和产量之间的关系 微核、凋亡核和持续的染色体重排 将会被确定。其次，由于辐射已知能在体内诱导碱性成纤维细胞生长因子 内皮细胞导致细胞凋亡的抑制，实验是 这将决定氦离子辐射是否会改变水平 HLE细胞中碱性成纤维细胞生长因子mRNA或蛋白的表达及其是否存在相关性 与细胞凋亡事件有关。最后，对可能的 氦离子辐照对晶状体细胞内蛋白质的修饰 建议。该计划将阐明氦-离子-之间的关系。 体外诱导HLE细胞染色质损伤及其生物学意义 对细胞在由此造成的损伤中幸存下来的后果。在修订后的 应用程序的重大变化包括：1)HLE来源增加 细胞，2)来自信使核糖核酸和蛋白质分析的新的初步数据(SDS-PAGE) 指数和融合细胞培养，3)新的初步数据 免疫学证据证实晶状体细胞表型的可行性 区分，4)一些修改的实验方案，5)和一个 对这一快速增长领域发表的文献进行了最新回顾。 这一知识将允许与可用的相关比较 体内的实验工作，并可能为潜在的变化提供基础 可以降低白内障风险的治疗方案。这个 导致白内障发生的生物机制也可能是 已澄清。

项目成果

Growth and differentiation of human lens epithelial cells in vitro on matrix.

• 发表时间: 2000-11
• 期刊: Investigative ophthalmology & visual science
• 影响因子: 4.4
• 作者: Eleanor A. Blakely;K. Bjornstad;Polly Y. Chang;M. P. McNamara;Edmund Chang;G. Aragon;Serena P. Lin;GeMing Lui;Jon R. Polansky

New measurements for hadrontherapy and space radiation: biology.

• 发表时间: 2001
• 期刊: Physica medica : PM : an international journal devoted to the applications of physics to medicine and biology : official journal of the Italian Association of Biomedical Physics
• 作者: E. Blakely

LAURISTON S. TAYLOR LECTURE ON RADIATION PROTECTION AND MEASUREMENTS: WHAT MAKES PARTICLE RADIATION SO EFFECTIVE?

• DOI: 10.1097/hp.0b013e31826a5b85
• 发表时间: 2012-11-01
• 期刊: HEALTH PHYSICS
• 影响因子: 2.2
• 作者: Blakely, Eleanor A.