Differentiation of Human ES and iPS Cells into Lens Cells

人 ES 和 iPS 细胞分化为晶状体细胞

• 批准号: 8044309
• 负责人: Ales Cvekl
• 金额: $ 23.94万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8044309
• 关键词: Differentiation; Human; ES; iPS; Cells

DESCRIPTION (provided by applicant): The long-term goal of this program is to develop new models to study human age-onset cataract via lens cells generated from embryonic stem (ES) cells. These models employ patient-derived induced pluripotent stem (iPS) cells differentiated into lens cells to study lens-cell homeostasis under conditions known to induce human cataractogenesis. The immediate goal of this R21 application is to perform systematic exploratory studies to generate lens cells form human embryonic stem (ES) cells using defined cell culture conditions. Age-related cataract is a disease of the lens that is responsible for nearly half of blindness worldwide, and is expected to increase as a result of increased life expectancy in both developed and underdeveloped countries. Age-onset cataracts develop after the age of 40 as a result of the progressive breakdown of the lens microarchitecture. Age-onset cataract is a complex disease involving both genetic and environmental factors. Lens opacities are thought to originate from the cumulative damage of environmental insults on lens proteins and other structural components of the lens from UV-B exposure, low antioxidant intake, and cigarette smoking. A systemic approach to study human cataract is hampered both by the lack of appropriate animal models, and limited use of primary lens cell and organ cultures. We have now identified an experimental protocol to generate large quantities of lens cells progenitor cells differentiated from human ES cell line, H1. This proposal will (1) establish an optimized experimental protocol to generate a highly enriched population of lens progenitor cells from human ES cells, (2) will establish conditions to generate differentiated lentoid bodies followed by their characterization, and (3) will test these experimental conditions using a representative pool of six iPS cell lines. These data will lay foundation for generation of lens cells using iPS cells from distinct cataract patients. The patient-derived lens cells will be available at quantities sufficient for biochemical studies of specific antioxidant defensive mechanisms to identify those lens protective mechanisms that play key roles in cataractogenesis. By identifying molecular targets for drugs to exploit, this novel approach will inspire a rational search for drugs to reduce the effects of aging on the lens. Collectively, these exploratory studies will be used for R01 projects to model human cataract using genetically and phenotypically characterized biological materials. PUBLIC HEALTH RELEVANCE: This application is relevant to human health as lens cataract is a major cause of worldwide blindness. Age- related cataract develops at some time after 40 years of age as a result of progressive breakdown of the ocular lens structure. Current treatment for senile cataract generally consists of surgery that replaces the opaque lens with an artificial intraocular lens. Although the surgery is performed routinely in the US at a rate of 1.8-2 million patients per annum, it represents a major Medicare reimbursement category. It has been estimated by the National Eye Institute, NIH (Bethesda, MD) that a 10-year delay in the onset of cataracts, could decrease the number of surgeries needed by almost one half, thus significantly decreasing vision care costs.

描述（由申请人提供）：该项目的长期目标是开发新模型，通过胚胎干（ES）细胞产生的晶状体细胞来研究人类老年性白内障。这些模型采用源自患者的诱导多能干（iPS）细胞分化为晶状体细胞，以研究已知会诱发人类白内障发生的条件下的晶状体细胞稳态。该 R21 应用的直接目标是进行系统的探索性研究，以使用规定的细胞培养条件从人类胚胎干 (ES) 细胞生成晶状体细胞。与年龄相关的白内障是一种晶状体疾病，导致全世界近一半的失明，并且由于发达国家和不发达国家预期寿命的增加，预计该病的数量会增加。由于晶状体微结构逐渐损坏，40 岁后会出现老年性白内障。老年性白内障是一种复杂的疾病，涉及遗传和环境因素。晶状体浑浊被认为是由于 UV-B 暴露、低抗氧化剂摄入量和吸烟等环境损伤对晶状体蛋白和晶状体其他结构成分的累积损害所致。研究人类白内障的系统方法因缺乏适当的动物模型以及原代晶状体细胞和器官培养物的使用有限而受到阻碍。我们现在已经确定了一种实验方案，可以产生大量从人类 ES 细胞系 H1 分化而来的晶状体细胞祖细胞。该提案将 (1) 建立一个优化的实验方案，以从人类 ES 细胞中产生高度富集的晶状体祖细胞群，(2) 将建立产生分化的晶状体体的条件，然后对其进行表征，(3) 将测试这些实验使用六种 iPS 细胞系的代表性池的条件。这些数据将为使用来自不同白内障患者的 iPS 细胞生成晶状体细胞奠定基础。来自患者的晶状体细胞的数量将足以用于特定抗氧化防御机制的生化研究，以确定在白内障发生中起关键作用的晶状体保护机制。通过确定药物的分子靶标，这种新颖的方法将激发人们合理地寻找药物，以减少老化对晶状体的影响。总的来说，这些探索性研究将用于 R01 项目，利用遗传和表型特征的生物材料模拟人类白内障。 公共健康相关性：该应用与人类健康相关，因为晶状体白内障是全球失明的主要原因。由于眼晶状体结构逐渐损坏，年龄相关性白内障在 40 岁后的某个时间出现。目前老年性白内障的治疗通常包括用人工晶状体代替不透明晶状体的手术。尽管这种手术在美国是常规手术，每年有 18-200 万名患者接受手术，但它代表了医疗保险的主要报销类别。据美国国立卫生研究院 (NIH) 国家眼科研究所（马里兰州贝塞斯达）估计，白内障发病延迟 10 年，所需手术数量可减少近一半，从而显着降低视力保健费用。