Chemical approaches for studying the biology of CD38

研究 CD38 生物学的化学方法

• 批准号: 8389598
• 负责人: Hening Lin
• 金额: $ 28.26万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8389598
• 关键词: Active Sites; Address; Adenosine Diphosphate Ribose; Affect; Affinity; Antibodies; Autistic Disorder; Binding; Biochemistry; Biological Markers; Biology; Biomedical Research; Cell Adhesion; Cell Culture Techniques; Cell Differentiation process; Cell Surface Proteins; Cell membrane; Cell physiology; Cell surface; Cells; Chemicals; Communities; Conflict (Psychology); Cyclic ADP-Ribose; Data; Defect; Dependence; Diabetes Mellitus; Differentiation Antigens; Disease; Endocytosis; Enzymes; Event; Fluorescent Probes; Goals; Hematopoietic; Immune; Knowledge; Label; Laboratories; Lead; Life; Ligand Binding; Ligands; Location; Mammalian Cell; Mediating; Methods; Mitogen-Activated Protein Kinases; Monitor; NAADP; NADP; Neuropeptides; Nicotinamide adenine dinucleotide; Organelles; Osteoporosis; PECAM1 gene; Pathway interactions; Permeability; Phosphorylation; Play; Poly(ADP-ribose) Polymerases; Production; Proteins; Receptor Activation; Reporting; Research Personnel; Role; Science; Second Messenger Systems; Serum; Signal Transduction; Sirtuins; Social Behavior; Streptavidin; Time; Up-Regulation; Work; analog; base; cell type; cytokine; design; disease diagnosis; extracellular; human disease; in vivo; insulin secretion; leukemia; prevent; public health relevance; receptor; receptor function; research study; response; second messenger; small molecule; tool; trafficking

DESCRIPTION (provided by applicant): The significance of understanding biomarkers to take full advantage of them in disease diagnosis and treatment as well as in biomedical research is well recognized by the science community. CD38, originally identified as a differentiation marker for hematopoietic cells, is present in many other types of cells, and its deregulation is found to contribute to several different human diseases, including leukemia, social behavior defects, diabetes, and osteoporosis. Understanding the roles CD38 plays in various normal and pathological conditions has the potential to offer new ways to treat these diseases. So far, some knowledge about CD38 has been acquired, though limited, thanks to the efforts from researchers in the field. CD38 acts both as an enzyme and a receptor. The enzymatic activity can convert nicotinamide adenine dinucleotide (NAD) to adenosine diphosphate ribose (ADPR) and cyclic ADPR (cADPR), and NAD phosphate (NADP) to nicotinic acid adenine dinucleotide phosphate (NAADP). Both cADPR and NAADP are potent Ca2+ messengers that can trigger Ca2+ release from internal stores. As a receptor, when activated by certain ligands, CD38 can trigger the phosphorylation of intracellular proteins, including c-Cbl and mitogen-activated protein kinases (MAPK). However, the exact roles CD38 plays in most normal and pathological conditions are poorly understood at present, despite the accumulation of the knowledge mentioned above. Several unanswered questions prevent a clear and complete understanding of CD38 function. For example, how CD38 gains access to its NAD or NADP substrate, whether CD38 is present in intracellular organelles, and how the receptor function affects the enzymatic function. In this proposal, we will develop and use chemical tools, particularly NAD analogs that can covalently label CD38 in live cells, to address these important questions concerning CD38 biology. These CD38 probes have unique features, such as cell permeability, compatibility with ligand binding to CD38, and the ability to inhibit CD38 enzymatic activity. These small molecule probes can be used to track CD38 trafficking in real time in live cells, determine the intracellular distribution of CD38, and isolate and identify unknown CD38 ligands. These experiments cannot be easily achieved using other methods. Our studies will lead to a more detailed mechanistic picture of CD38 biochemistry, which will help to understand the function of CD38 in various normal and pathological conditions and potentially lead to new ways to treat diseases involving CD38, such as leukemia, diabetes, autism, and osteoporosis. In addition, the NAD metabolizing capability of CD38 can have a significant impact on other NAD-dependent enzymes, particularly the NAD-dependent deacetylases (sirtuins) and poly(ADP-ribose) polymerases (PARPs). Understanding how CD38 works should shed lights on the potential interactions between CD38 and other NAD-dependent enzymes.

描述（由申请人提供）：理解生物标志物在疾病诊断和治疗以及生物医学研究中充分利用它们的意义得到了科学界的认可。 CD38最初被鉴定为造血细胞的分化标记物，存在于许多其他类型的细胞中，并且发现其失调会导致几种不同的人类疾病，包括白血病，社会行为缺陷，糖尿病和骨质疏松症。了解CD38在各种正常和病理条件下的作用有可能提供新方法来治疗这些疾病。到目前为止，由于该领域的研究人员的努力，尽管有限，但已经获得了有关CD38的一些知识。 CD38既充当酶和受体。酶活性可以将烟酰胺腺嘌呤二核苷酸（NAD）转换为二磷酸二磷酸核糖（ADPR）和环状ADPR（CADPR），以及NAD磷酸盐（NADP）将其转换为烟碱酸腺嘌呤二核苷酸（NAADP）。 CADPR和NAADP都是有效的CA2+ Messenger，可以触发内部商店的Ca2+释放。作为受体，当通过某些配体激活时，CD38会触发细胞内蛋白的磷酸化，包括C-CBL和有丝分裂原激活的蛋白激酶（MAPK）。然而，尽管上述知识的积累，但目前，CD38在大多数正常情况下的确切作用在大多数正常情况下都尚未理解。几个未解决的问题阻止了对CD38功能的清晰完整的了解。例如，CD38如何获得对其NAD或NADP底物的访问，是否存在CD38在细胞内细胞器中，以及受体功能如何影响酶促功能。在此提案中，我们将开发和使用化学工具，尤其是可以在活细胞中共价标记CD38的NAD类似物，以解决有关CD38生物学的这些重要问题。这些CD38探针具有独特的特征，例如细胞渗透性，与配体与CD38结合的兼容性以及抑制CD38酶活性的能力。这些小分子探针可用于实时跟踪CD38运输，确定CD38的细胞内分布，并分离并鉴定未知的CD38配体。这些实验无法使用其他方法轻松实现。我们的研究将导致CD38生物化学的更详细的机理图片，这将有助于了解CD38在各种正常和病理条件下的功能，并有可能导致治疗涉及CD38的疾病的新方法，例如白血病，糖尿病，自闭症，自闭症和骨质疏松症。另外，CD38的NAD代谢能力可以对其他NAD依赖性酶产生重大影响，尤其是NAD依赖性脱乙酰基酶（Sirtuins）和聚（ADP-核糖）聚合酶（PARPS）。了解CD38的工作原理应阐明CD38与其他NAD依赖性酶之间的潜在相互作用。