Structural and molecular determinants of protein phosphatase 2A function in lung cancer

肺癌中蛋白磷酸酶 2A 功能的结构和分子决定因素

基本信息

批准号：
10675070
负责人：
Goutham Narla
金额：
$ 62.48万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10675070
关键词：
Anti-Cholinergics Antibodies Antipsychotic Agents Binding Biological Biology C-terminal CRISPR/Cas technology Catalytic Domain Cell Line Cessation of life Characteristics Chemicals Clinical Collection Combined Modality Therapy Complex Coupling Data Diagnosis Disease Dose Limiting Engineering Enzymes Genetic Goals Holoenzymes Human Immunoprecipitation In Vitro Incidence Individual K-ras mouse model KRAS2 gene Libraries Lung Adenocarcinoma MEK inhibition Malignant Neoplasms Malignant neoplasm of lung Mass Spectrum Analysis Missense Mutation Modeling Molecular Mus Mutation Non-Small-Cell Lung Carcinoma Nude Mice Oncogenes Oncogenic Patients Periodicity Pharmaceutical Preparations Pharmacology Phenothiazines Phenotype Phosphoric Monoester Hydrolases Physiological Post-Translational Protein Processing Post-Translational Regulation Prognosis Property Protein Dephosphorylation Protein Phosphatase Inhibitor Protein phosphatase Recurrent tumor Regulation Resistance development Role Signal Transduction Somatic Mutation Specificity Structure Survival Rate Tail Therapeutic Transgenic Mice Tumor Promotion Tumor Suppressor Proteins Tumor-Derived Woman anticancer activity cancer therapy cell immortalization combinatorial complement C2a design effective therapy in vitro activity in vivo in vivo Model inhibitor insight kinase inhibitor loss of function lung cancer cell lung carcinogenesis men mortality mutant next generation novel overexpression pre-clinical protein activation protein phosphatase 2A regulatory subunit 65 kDa response scaffold small molecule tool treatment response treatment strategy tumor tumor progression

项目摘要

Project Summary/Abstract Lung cancer progression involves coordinate changes in both oncogene and tumor suppressor function. Protein Phosphatase 2A (PP2A) is a tumor suppressor that is dysregulated and deactivated in lung cancer. PP2A is inactivated through several mechanisms including somatic mutations, suppression of individual subunits, increased expression of endogenous PP2A inhibitors and changes in post-translational modifications of the C subunit. While several therapeutics have been developed to indirectly target PP2A, recent studies have uncovered small molecules such as phenothiazines that directly activate PP2A. Nevertheless, because their pronounced extrapyramidal and anti-cholinergic effects were severely dose limiting, further pursuit of phenothiazines and related dibenzazepines for treatment of cancer seems unlikely. Our lab has reengineered the tricyclic neuroleptics to decouple the CNS pharmacology from the anti-proliferative properties of this drug- class. We have developed and characterized these small molecule activators of PP2A (SMAPs) as both chemical tools to probe PP2A regulation and for the treatment of PP2A dependent diseases. Our combined structure- function studies have revealed that SMAPs bind to PP2A in a way that protects the regulatory C-terminal tail of the catalytic subunit and stabilizes the holoenzyme. Our goal is to define the SMAP induced alterations on PP2A holoenzyme composition that promote the tumor suppressive properties of PP2A. We aim to identify the tumor suppressive PP2A regulatory subunits enhanced by SMAPs in order to correlate treatment response to clinical and genetic characteristics, and to aid in the design of more selective, next-generation SMAPs. In this project, three robust aims are designed to assess how activation of PP2A using SMAPs represents an effective treatment option for patients diagnosed with NSCLC. Aim 1 will structurally and functionally characterize the physiologic and cancer-relevant posttranslational regulation of PP2A by SMAPs. We will investigate the SMAPs-specific structural modulation of the PP2A holoenzyme by various, recurrent tumor derived mutations through coupling methyl-PP2A-C antibody and immunoprecipitation-mass spectrometry based approaches to quantify post- translational modification and PP2A activity. Aim 2 will discern how PP2A Aα mutations modulate the biological activity of SMAPs. We will assess the functional role of common missense mutations in lung cancer by using isogenic CRISPR/Cas9 cell lines to identify cellular phenotypes, subunit expression, SMAPs response, and biological responses in nude mice. Aim 3 will determine combination treatment strategies to promote regression of tumors in KRAS mutant lung cancer. We will examine rational treatment combinations with SMAPs based on our studies demonstrating that protein phosphatase 2A (PP2A) activity defines the response of KRAS mutant lung cancer cells across library of >200 kinase inhibitors. Collectively, these studies will reveal novel insights into the molecular basis of regulation of PP2A and functional consequences of PP2A activation/inactivation in lung cancer.

项目概要/摘要肺癌的进展涉及癌基因和抑癌蛋白功能的协调变化。磷酸酶 2A (PP2A) 是一种肿瘤抑制因子，在肺癌中 PP2A 失调并失活。通过多种机制失活，包括体细胞突变、抑制单个亚基、内源性 PP2A 抑制剂的表达增加以及 C 翻译后修饰的变化虽然已经开发了几种间接靶向 PP2A 的疗法，但最近的研究表明尽管如此，还是发现了吩噻嗪等小分子可以直接激活 PP2A。明显的锥体外系和抗胆碱能作用严重限制剂量，进一步追求我们的实验室已经重新设计了吩噻嗪类药物和相关的二苯并氮杂卓类药物用于治疗癌症的可能性。三环类精神安定药将中枢神经系统药理学与该药物的抗增殖特性分开- 我们开发了这些 PP2A (SMAP) 小分子激活剂，并将其表征为化学物质。探索 PP2A 调节和治疗 PP2A 依赖性疾病的工具我们的组合结构- 功能研究表明，SMAP 与 PP2A 结合，可保护调节性 C 末端尾部我们的目标是确定 SMAP 诱导的 PP2A 改变。促进 PP2A 肿瘤抑制特性的全酶组合物我们的目标是鉴定肿瘤。 SMAP 增强了抑制性 PP2A 调节亚基，以便将治疗反应与临床相关联和遗传特征，并帮助设计更具选择性的下一代 SMAP。设计了三个强有力的目标来评估使用 SMAP 激活 PP2A 如何代表一种有效的治疗目标 1 将从结构和功能上表征 NSCLC 患者的选择。我们将研究 SMAP 特异性的 PP2A 和癌症相关的翻译后调节。 PP2A全酶的结构调节是通过偶联产生的各种、复发性肿瘤衍生突变甲基-PP2A-C 抗体和基于免疫沉淀质谱的方法来量化后目标 2 将了解 PP2A Aα 突变如何调节生物学。我们将使用 SMAP 的活性来评估常见错义突变在肺癌中的功能作用。同基因 CRISPR/Cas9 细胞系，用于鉴定细胞表型、亚基表达、SMAP 反应和目标 3 将确定促进消退的联合治疗策略。我们将基于 SMAP 来研究合理的治疗组合。我们的研究表明，蛋白磷酸酶 2A (PP2A) 活性决定了 KRAS 突变体的反应总的来说，这些研究将揭示超过 200 种激酶抑制剂库中的肺癌细胞的新见解。肺中 PP2A 调节的分子基础以及 PP2A 激活/失活的功能后果癌症。

项目成果

期刊论文数量（16）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Small molecule inhibitors of 15-PGDH exploit a physiologic induced-fit closing system.

15-PGDH 的小分子抑制剂利用生理诱导贴合闭合系统。

DOI：
发表时间：
2023-02-11
期刊：
影响因子：
16.6
作者：
Huang, Wei;Li, Hongyun;Kiselar, Janna;Fink, Stephen P;Regmi, Sagar;Day, Alexander;Yuan, Yiyuan;Chance, Mark;Ready, Joseph M;Markowitz, Sanford D;Taylor, Derek J
通讯作者：
Taylor, Derek J

Protein phosphatase 2A controls ongoing DNA replication by binding to and regulating cell division cycle 45 (CDC45).

蛋白磷酸酶 2A 通过结合并调节细胞分裂周期 45 (CDC45) 来控制正在进行的 DNA 复制。

DOI：
发表时间：
2019
期刊：
影响因子：
0
作者：
Perl, Abbey L;O'Connor, Caitlin M;Fa, Pengyan;Mayca Pozo, Franklin;Zhang, Junran;Zhang, Youwei;Narla, Goutham
通讯作者：
Narla, Goutham

SLX4IP N-terminus dictates telomeric localization in ALT-like castration-resistant prostate cancer cell lines.

SLX4IP N 末端决定 ALT 样去势抵抗性前列腺癌细胞系中的端粒定位。

DOI：
发表时间：
2021-11
期刊：
影响因子：
0
作者：
Mangosh, Tawna L;Grabowska, Magdalena M;Taylor, Derek J
通讯作者：
Taylor, Derek J

Cryo-Electron Microscopy Structure of an Acinetobacter baumannii Multidrug Efflux Pump.

鲍曼不动杆菌多药外排泵的冷冻电子显微镜结构。

DOI：
发表时间：
2019
期刊：
影响因子：
6.4
作者：
Su, Chih;Morgan, Christopher E;Kambakam, Sekhar;Rajavel, Malligarjunan;Scott, Harry;Huang, Wei;Emerson, Corey C;Taylor, Derek J;Stewart, Phoebe L;Bonomo, Robert A;Yu, Edward W
通讯作者：
Yu, Edward W

Targeting protein phosphatase PP2A for cancer therapy: development of allosteric pharmaceutical agents.

靶向蛋白磷酸酶 PP2A 用于癌症治疗：变构药物制剂的开发。