GLOBIN EXPRESSION IN AN ERYTHROID PROGENITOR CULTURE SYSTEM

红细胞祖细胞培养系统中的珠蛋白表达

• 批准号: 2572909
• 负责人: A N SCHECHTER
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 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/2572909
• 关键词: Primates; blood disorder chemotherapy; clinical research; combination chemotherapy; cyclic compound; drug screening /evaluation; erythroid stem cell; erythropoiesis; fatty acids; gene expression; globin; heme; hemoglobin F; hemoprotein biosynthesis; human therapy evaluation; human tissue; hydroxyurea; sickle cell anemia; thalassemia; tissue /cell culture

Increased fetal hemoglobin (HbF) in cells of patients with b- hemoglobinopathies (b-thalassemia and sickle cell anemia) ameliorates the clinical symptoms of the underlying disease. Several pharmacologic agents have been used to stimulate HbF synthesis; e.g., treatment of patients with hydroxyurea resulted in higher HbF levels and a significant improvement in the clinical symptoms. Yet, there is a considerable interest in other, less toxic agents with even higher potential to increase HbF. So far only a handful of agents have been tested, mainly due to the lack of an appropriate experimental system that allows a rapid and accurate determination of the effect on relevant cells. We have developed a two-phase liquid culture system for growing erythroid progenitors derived from the peripheral blood of normal individuals and patients with hemoglobinopathies. We showed that the system recapitulates many aspects of erythropoiesis in vivo, including stimulation of HbF production by pharmacologic agents. The purpose of our research is to utilize this culture procedure and various biochemical, molecular and immunological methods to study the regulation of Hb production and for screening of agents for their HbF- stimulating potential with the final objective to produce an efficient, low toxicity therapy. The experiments include optimization of the conditions for stimulation of HbF production, followed by screening of various agents (including derivatives of hydroxyurea, aromatic fatty acids, hemoglobin and hemin) and subsequently studying their mode(s) of action. The latter studies will provide the rationale for further screening of additional agents and for their chemical modification in order to increase efficacy. Classification of the drugs according to functional and mechanistic considerations will enhance experiments with combinations of drugs belonging to different groups and, thus, expected to act synergistically. In addition, we are attempting to develop, based on the cell culture system, an assay for evaluating an individual patient's response to a particular drug or drug combination. We are also working with a newly developed robotic system for screening large numbers of compounds which may affect globin gene transcription; chemical libraries of thousands of compounds are now being screened by this method. We are testing compounds which score positively in this system in our culture system, as well as in rhesus primates (in conjunction with the NHLBI facility).

B-型糖尿病患者细胞中胎儿血红蛋白（HbF）增加， 血红蛋白病（b-地中海贫血和镰状细胞贫血）改善 潜在疾病的临床症状。 几 已经使用药理学试剂来刺激HbF合成;例如， 使用羟基脲治疗患者导致HbF水平升高 临床症状明显改善。 然而， 对其他毒性更低、毒性更高的药物也有相当大的兴趣。 有可能增加HbF。 到目前为止，只有少数特工 测试，主要是由于缺乏适当的实验系统 这使得能够快速准确地确定对 相关细胞 我们开发了一种两相液体培养系统 用于生长来源于外周血的红系祖细胞， 正常个体和血红蛋白病患者。 我们展示 该系统概括了体内红细胞生成的许多方面， 包括通过药物刺激HbF产生。 的 我们研究的目的是利用这种培养方法， 各种生物化学、分子和免疫学方法来研究 调节Hb产生和筛选用于其HbF的试剂， 激发潜力，最终目标是 高效低毒的治疗方法 实验包括优化 刺激HbF产生的条件，然后 筛选各种试剂（包括羟基脲的衍生物， 芳香族脂肪酸，血红蛋白和氯化血红素），并随后研究 他们的行动方式。 后一项研究将提供 进一步筛选其他药物及其 化学修饰以提高功效。 分类 根据功能和机制考虑， 将加强实验的药物组合属于 不同的群体，因此，预计将发挥协同作用。 在 此外，我们正在尝试开发基于细胞培养的 系统，一种用于评估个体患者对药物的反应的测定， 特定药物或药物组合。我们还与一个新的 开发了用于筛选大量化合物的机器人系统 其可能影响珠蛋白基因转录; 目前正在用这种方法筛选数以千计的化合物。 我们 在我们的文化中测试在这个系统中得分为正的化合物 系统，以及恒河猴灵长类动物（与NHLBI 设施）。