Screen and Design p18 Chemical Probes for Hematopoietic Stem Cell Self-Renewal

用于造血干细胞自我更新的 p18 化学探针的筛选和设计

• 批准号: 8284383
• 负责人: Xiang-Qun Xie
• 金额: $ 18.74万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8284383
• 关键词: Affect; Behavior; Binding; Biochemical; Biological Assay; Bone Marrow; CDK4 gene; CDK6-associated protein p18; Cell Cycle; Cell Therapy; Cells; Chemicals; Chemistry; Clinic; Collaborations; Colony-Forming Units Assay; Commit; Computer Simulation; Cyclin-Dependent Kinase Inhibitor; Data; Databases; Disease; Drug Delivery Systems; Ectopic Expression; Eligibility Determination; Equilibrium; Figs - dietary; Future; G1 Phase; Goals; Grant; Harvest; Hematological Disease; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; Immune system; In Vitro; Laboratories; Lead; Libraries; Malignant - descriptor; Malignant Neoplasms; Mediating; Modification; Molecular Target; Mus; Nature; Outcome; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Protein Binding; Proteins; Protocols documentation; Published Comment; Publishing; Recommendation; Regenerative Medicine; Research; Research Design; Research Proposals; Retroviridae; Screening procedure; Signal Transduction; Site-Directed Mutagenesis; Staging; Stem Cell Research; Stem cell transplant; Stem cells; Stimulus; Study models; Synthesis Chemistry; Techniques; Testing; Therapeutic; Therapeutic Uses; Toxic effect; Umbilical Cord Blood; Work; adult stem cell; base; design; high risk; human stem cells; improved; in vitro Bioassay; in vivo; inhibitor/antagonist; innovation; leukemogenesis; meetings; member; novel; novel strategies; research and development; research study; response; self-renewal; small molecule; stem cell biology; stem cell therapy; virtual

Human hematopoietic stem cells (HSC) transplantation is currently being used as regenerative medicine for the treatment of congenital deficiencies and malignant diseases as well as cancers and other disorders of the blood and immune systems. However, despite all the enthusiasm surrounding HSC biology and therapeutics, the potential of HSC-based therapies has yet to be fully realized. A major roadblock of broader use of the adult stem cells is the limited number of HSC per harvest for therapeutic benefit and their poorly understood expansion and differentiation behavior in response to proliferative stimuli. In vitro expansion of HSC remains a major challenge for wide applications of HSC transplantation for patients. Our studies show that p18, a member of the cyclin-dependent kinase (CDK) inhibitors (CKI), is a potent negative regulator of HSC self- renewal. Thus, we hypothesize that p18 is a unique drug target, and that small molecules capable of blocking p18 function and interfering with p18/CDK6 interactions are likely to be potent drugs for activating HSC self- renewal. Our objective is to screen/identify p18 inhibitors that act by disrupting p18/CDK6 interactions, thus activating HSC self-renewal and increasing the quantity of active stem cells, and to use them as chemical probes for mechanism studies of HSC self-renewal. The feasibility of the proposed innovative research is supported by the proof-of-principle pilot data obtained by well-established research teams that have complementary expertise for the proposed research. Considering the limited throughput capacity of the current HSC bone-marrow culture protocol, we propose first to use our established in silico screening approach for initial screening to generate p18-focused lead sublibraries (Aim 1). We also apply NMR assays to screen/validate and characterize the p18 hits and their binding interactions with the protein in order to generate p18-active subsets (Aim 2A). Also, the small subsets of validated p18-targeting compounds will then be confirmed by extensive HSC functional assays (Aim 2B). Through these, the compounds that are capable of increasing the number of active stem cells in the bone-marrow culture will be identified as leads. The discovered leads are then used as specific chemical probes for studies of p18/CDK6 interactions and signaling mechanisms of the G1-phase of the cell cycle. As a future plan, the identified leads will be further optimized by chemistry modification and SAR medicinal chemistry studies to improve the potency and cell toxicity. Our long- term goal is to identify/design CKI p18-specific small molecule effectors that can either maintain/stimulate self- renewal of hematopoietic stem cells in a predictable manner, and ultimately to develop new drugs for HSC therapies. Achieving this goal will have a significant impact on stem cell drug research development in general.

人类造血干细胞（HSC）移植目前被用作再生医学 先天性缺陷和恶性疾病以及癌症和其他疾病的治疗 血液和免疫系统。然而，尽管人们对 HSC 生物学和治疗充满热情， 基于 HSC 的疗法的潜力尚未完全实现。更广泛使用成人的主要障碍 干细胞是指每次收获的造血干细胞数量有限，无法发挥治疗作用，但人们对它们知之甚少 响应增殖刺激的扩张和分化行为。 HSC 的体外扩增仍然是一个 HSC移植在患者中广泛应用的主要挑战。我们的研究表明，p18 细胞周期蛋白依赖性激酶 (CDK) 抑制剂 (CKI) 的成员，是 HSC 自身的有效负调节因子。 更新。因此，我们假设 p18 是一个独特的药物靶点，并且小分子能够阻断 p18 功能和干扰 p18/CDK6 相互作用可能是激活 HSC 自身的有效药物。 更新。我们的目标是筛选/鉴定通过破坏 p18/CDK6 相互作用发挥作用的 p18 抑制剂，从而 激活HSC自我更新，增加活性干细胞的数量，并将其用作化学物质 HSC自我更新机制研究的探针。所提出的创新研究的可行性是 得到了由成熟的研究团队获得的原理验证试点数据的支持，这些研究团队已经 拟议研究的补充专业知识。考虑到当前吞吐能力有限 HSC 骨髓培养方案，我们建议首先使用我们建立的计算机筛选方法 初步筛选以生成以 p18 为重点的先导子文库（目标 1）。我们还应用 NMR 分析来 筛选/验证并表征 p18 命中及其与蛋白质的结合相互作用，以便生成 p18 活性子集（目标 2A）。此外，经过验证的 p18 靶向化合物的小子集将被 通过广泛的 HSC 功能测定证实（目标 2B）。通过这些，化合物能够 增加骨髓培养物中活性干细胞的数量将被确定为先导。这 然后将发现的先导化合物用作特定化学探针，用于研究 p18/CDK6 相互作用和信号传导 细胞周期 G1 期的机制。作为未来的计划，已确定的线索将进一步优化 化学修饰和 SAR 药物化学研究，以提高效力和细胞毒性。我们的长期 长期目标是识别/设计 CKI p18 特异性小分子效应器，该效应器可以维持/刺激自我 以可预测的方式更新造血干细胞，最终开发针对 HSC 的新药 疗法。实现这一目标将对干细胞药物研究的总体发展产生重大影响。

项目成果

Cancer genomics: opportunities for medicinal chemistry?

癌症基因组学：药物化学的机会？

• DOI: 10.4155/fmc.16.1
• 发表时间: 2016
• 期刊: Future medicinal chemistry
• 影响因子: 4.2
• 作者: Wang,Lirong;Xie,Xiang-Qun
• 通讯作者: Xie,Xiang-Qun

N-terminal arginylation generates a bimodal degron that modulates autophagic proteolysis.

N 端精氨酸化产生双峰降解决定子，调节自噬蛋白水解。

• DOI: 10.1073/pnas.1719110115
• 发表时间: 2018
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Yoo,YoungDong;Mun,SuRan;Ji,ChangHoon;Sung,KiWoon;Kang,KeumYoung;Heo,AhJung;Lee,SuHyun;An,JeeYoung;Hwang,Joonsung;Xie,Xiang-Qun;Ciechanover,Aaron;Kim,BoYeon;Kwon,YongTae
• 通讯作者: Kwon,YongTae