Impact of attenuated oxidative damage in aged hematopoietic stem cells

氧化损伤减弱对衰老造血干细胞的影响

• 批准号: 9770203
• 负责人: Isabel Beerman
• 金额: $ 5.36万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9770203
• 关键词: Acetylcysteine; Address; Age; Antioxidants; Attenuated; Biological Assay; Bone Marrow; Cell Aging; Cell Compartmentation; Cells; DNA Damage; Ensure; Goals; Hematopoietic stem cells; Intervention; Laboratories; Life; Link; Mus; Stem cells; Testing; Tissues; Transplantation; aged; cohort; congenic; drinking water; experimental study; functional loss; improved; insight; male; oxidative damage; peripheral blood; reconstitution

To establish if there are reduced levels of damage in the HSC compartment after N-acetylcysteine treatment, we administered 1mg/mL of NAC in the drinking water to young and aged C57BL/6 male mice. Age-matched non-treated control cohorts were also included in the experiment. Initial FLARE comet experiments assaying for DNA damage demonstrated a significant decrease in the levels of oxidative damage (FPG+) in old HSCs after one month of NAC treatment. We examined the bone marrow for changes in the primitive bone marrow compartments, but do not see any significant differences in the primitive bone marrow compartments assayed. Functional assays of NAC treated HSCs by transplant assays demonstrated no significant changes in functional potential of HSCs with reduced oxidative damage in peripheral blood (Old-NAC vs Old-WT) or in bone marrow analysis in the primary transplant. To test long term potential of HSCs, secondary transplants were performed on 200 donor derived HSCs again 2x105 congenic competitor cells. Peripheral blood analysis again showed no significant differences in either total reconstitution or in lineage composition four months post-secondary transplant. Current efforts are focused on establishing if aged HSC function can be improved by supplementing NAC during the transplant setting, where the HSCs will be actively dividing, and antioxidant benefits may be more pronounced in this setting.

为了确定N-乙酰半胱氨酸治疗后HSC隔室的损伤水平降低，我们对饮用水中的NAC施用了1mg/mL的NAC，以对年轻和年龄的C57BL/6雄性小鼠施用。实验中还包括年龄匹配的未处理对照组。在NAC治疗一个月后，对DNA损伤进行测定的初始耀斑彗星实验表明，旧HSC的氧化损伤水平（FPG+）显着降低。我们检查了骨髓是否有原始骨髓室的变化，但没有看到测定的原始骨髓室的任何显着差异。 通过移植测定对NAC处理的HSC进行的功能测定表明，HSC的功能潜力没有显着变化，而外周血（Old-NAC与Old-WT）的氧化损伤或主要移植中的骨髓分析没有显着变化。为了测试HSC的长期潜力，对200个供体衍生的HSC进行了继发性移植，再次进行了2x105个先天性竞争者细胞。 周围血液分析再次显示出四个月的次生移植后四个月的总重建或谱系组成均无显着差异。当前的努力集中在建立是否可以通过在移植设置中补充NAC来提高老化的HSC功能，而HSC将积极分裂，并且在此环境中可能会更明显。