Cancer cell signaling through lipids complexed to proteins

通过脂质与蛋白质复合的癌细胞信号传导

基本信息

批准号：
8708521
负责人：
Raymond Daniel Blind
金额：
$ 11.22万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-11 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8708521
关键词：
1-Phosphatidylinositol 3-Kinase Accounting Acute Myelocytic Leukemia Adult African American American Cancer Society Antineoplastic Agents Binding Biochemistry Cancer Biology Cancer cell line Cell Nucleus Cells Cellular biology Chemicals Colorectal Cancer Complex Crystallography Data Deuterium Development Endometrial Carcinoma Enzymatic Biochemistry Enzyme Kinetics Enzymes Genetic Programming Hispanics Holly Human Human Cell Line Hydrogen In Vitro Incidence Intestinal Cancer Link Lipid Binding Lipids Malignant Neoplasms Malignant neoplasm of liver Malignant neoplasm of pancreas Manuscripts Maps Mediating Membrane Mentored Research Scientist Development Award Mentors Methods Molecular Molecular Structure Multienzyme Complexes Mutation Nuclear Nuclear Protein Nuclear Proteins Nuclear Receptors PTEN gene Pathway interactions Patients Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols Phospholipids Phosphoric Monoester Hydrolases Phosphotransferases Physiology Protein Chemistry Proteins Reporting Research Research Personnel Risk Roentgen Rays Role Science Signal Transduction Structure Surface Therapeutic Tissues Training Tumor Suppressor Proteins United States Woman Work X-Ray Crystallography anticancer research base cancer cell enzyme structure enzyme substrate enzyme substrate complex essential phospholipids experience high reward high risk improved inhibitor/antagonist inositol polyphosphate multikinase interfacial killings novel programs protein protein interaction simulation small molecule structural biology success tool transcription factor tumor tumor progression tumorigenesis tumorigenic

项目摘要

DESCRIPTION (provided by applicant): The American Cancer Society's 2012 report revealed that incidence rates are decreasing or stable for most cancers in the United States since 1999. Some notable exceptions are liver cancer in African Americans and Hispanics, endometrial cancer in African American women, colorectal cancer in patients under 50, and pancreatic cancer in every demographic. In all these tissues, NR5A nuclear receptors (NR5A1 and NR5A2) mediate genetic programs that are essential determinants of development, differentiation and adult function. At the molecular level, these two transcription factors bind phosphoinositides (PIPs), phospholipids that are essential to PI-3 kinase signaling in PTEN-dependent cancers. However almost nothing is known about how these lipids regulate NR5A transcriptional functions, or how dysregulation of those mechanisms contributes to the cancers mentioned above. Based on a manuscript currently in revision at Science, we hypothesize that PIP lipids bound to NR5As (and other nuclear proteins) are directly remodeled by the PI3- kinase inositol polyphosphate multikinase (IPMK) and the PTEN lipid phosphatase in cancer cells. This hypothesis is a clear departure from the standard dogma that PI3- kinases & PTEN only act on phosphoinositides in membranes. In Aim 1, we will determine how currently available small molecules inhibit IPMK using enzymology and crystallography, accelerating improvements of these inhibitors and training the PI in crystallography, X-ray diffraction methods and molecular structure determination, taking advantage of a relatively low-risk project. In Aim 2, we will use mutational analyses to map the interface between NR5A1 and IPMK, and attempt to crystallize NR5A/PIP/IPMK to determine the interfacial structure of this complex. This high-risk high-reward structure is hedged by Aim 1. In Aim 3, we will identify novel protein/PIP complexes that are substrates of PTEN and IPMK, using hypothesis-driven biochemistry. The candidate has had extensive training in protein chemistry, enzymology, biochemistry and cell biology, but has not had any training in structural biology. The mentor Holly Ingraham and advisory team have extensive experience in structural biology, particularly Robert Fletterick in NR5A structure and its links to pancreatic cancer, and Natalia Jura in kinase structure and membrane cancer biology. The candidate will expand his research program as an independent investigator with the training afforded by this K01 award, opening new avenues to cancer research.

描述（由申请人提供）：美国癌症协会 2012 年报告显示，自 1999 年以来，美国大多数癌症的发病率都在下降或保持稳定。一些值得注意的例外是非洲裔美国人和西班牙裔美国人的肝癌、非洲裔美国妇女的子宫内膜癌、 50 岁以下患者的结直肠癌，以及各个人群的胰腺癌。在所有这些组织中，NR5A 核受体（NR5A1 和 NR5A2）介导遗传程序，这些遗传程序是发育、分化和成年功能的重要决定因素。在分子水平上，这两种转录因子与磷酸肌醇 (PIP) 结合，这种磷脂对于 PTEN 依赖性癌症中的 PI-3 激酶信号转导至关重要。然而，关于这些脂质如何调节 NR5A 转录功能，或者这些机制的失调如何导致上述癌症，人们几乎一无所知。根据《Science》杂志目前正在修订的一篇手稿，我们假设与 NR5As（和其他核蛋白）结合的 PIP 脂质是由癌细胞中的 PI3 激酶肌醇多磷酸激酶 (IPMK) 和 PTEN 脂质磷酸酶直接重塑的。这一假设明显背离了 PI3-激酶和 PTEN 仅作用于膜中的磷酸肌醇的标准教条。在目标 1 中，我们将利用酶学和晶体学确定目前可用的小分子如何抑制 IPMK，加速这些抑制剂的改进，并利用相对低风险的项目对 PI 进行晶体学、X 射线衍射方法和分子结构测定方面的培训。在目标 2 中，我们将使用突变分析来绘制 NR5A1 和 IPMK 之间的界面图谱，并尝试使 NR5A/PIP/IPMK 结晶以确定该复合物的界面结构。这种高风险高回报的结构由目标 1 来对冲。在目标 3 中，我们将使用假设驱动的生物化学来鉴定作为 PTEN 和 IPMK 底物的新型蛋白质/PIP 复合物。该候选人接受过蛋白质化学、酶学、生物化学和细胞生物学方面的广泛培训，但没有接受过任何结构生物学方面的培训。导师Holly Ingraham和顾问团队在结构生物学方面拥有丰富的经验，特别是Robert Fletterick在NR5A结构及其与胰腺癌的联系方面的经验，以及Natalia Jura在激酶结构和膜癌生物学方面的经验。候选人将通过 K01 奖项提供的培训扩大其作为独立研究者的研究项目，为癌症研究开辟新途径。