Maxwell RSC rapid DNA/RNA purification instrument for high-quality genotyping and phenotyping research

Maxwell RSC 快速 DNA/RNA 纯化仪，用于高质量基因分型和表型研究

• 批准号: 10799221
• 负责人: Ying Yan
• 金额: $ 3.41万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10799221
• 关键词: Achievement; Anchorage-Independent Growth; Automobile Driving; CUL1 gene; Catalytic Domain; Cells; Communities; Complex; Critical Pathways; Cytoplasm; DNA; Data; Development; Disease; Equipment; F-Box Proteins; Family; Funding; Genes; Genotype; Goals; Human; Knowledge acquisition; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Medical center; Methods; Mus; Nebraska; Neoplasm Metastasis; Nucleic Acids; Oncogenic; Pathway interactions; Patients; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Protein Kinase; Protein Phosphatase 2A Regulatory Subunit PR53; Protein phosphatase; Proteins; RNA; RNA purification; Regulation; Request for Applications; Research; Research Project Grants; Retinal blind spot; Role; Sampling; Signal Transduction; Source; Specimen; Standardization; Substrate Specificity; Survival Rate; TP53 gene; Testing; Tissue Sample; Tissues; Tumor Suppression; Tumorigenicity; Universities; Work; cancer initiation; improved; innovation; instrument; knock-down; mouse model; new therapeutic target; novel; pancreatic cancer cells; scaffold; success; targeted treatment; tumor progression; tumorigenesis; ubiquitin-protein ligase

Abstract Pancreatic cancer (PC) is one of the most lethal cancers in the U.S. with a 5-year survival rate of 7%, which has essentially unchanged for the past 40 years. Despite extensive research, there are no effective targeted therapies for this disease, suggesting an insufficient understanding of oncogenic signaling networks in PC. While almost all of the signaling networks implicated in cancer rely on the cooperation of kinases and phosphatases to transduce signals promoting cancer, most efforts have only been put on the study of protein kinases in cancer while leaving protein phosphatases as a blind spot. PP2A is a family of heterotrimers that serve as the main source of Ser/Thr phosphatase activities in human cells, with each consisting of a scaffold subunit (Aα/β), a catalytic subunit (Cα/β), and one of many non-conserved regulatory (B) subunits (>27) that control the substrate specificities of PP2A complexes. Our preliminary studies identified PR55α, a regulatory subunit of PP2A, as an important new player in promoting malignant PC. In particular, our studies revealed that PR55 inhibits the MOB1/LATS auto-activation loop of the Hippo tumor suppression pathway, whose function is to induce the phosphorylation of YAP causing its cytoplasmic retention and proteasomal degradation. In line with the essential role of YAP signaling in Kras-driven PC development and progression, knockdown of PR55α impeded the anchorage-independent growth, tumorigenicity, and metastasis of PC cells. Our preliminary data also revealed that the stability of PR55α protein is negatively regulated by the tumor suppressor p53 and its target gene FBXL20, a substrate adaptor for the SCF (SKP1-CUL1-F-box protein) E3 ligase. Consistently, we observed that PR55α is elevated in PC specimens, where its expression correlates with poor patient survival. Based on these findings, we hypothesize that loss of p53 function results in the stabilization of PR55α protein, which, in turn, activates the YAP oncogenic signaling required for Kras-driving PC initiation and progression. We will test the hypothesis by three specific aims: Aim 1 to define the mechanism by which PR55α/PP2A promotes YAP activation and the significance of this cascade in oncogenesis and malignant potential of PC, Aim 2 to elucidate how p53 suppresses the level of PR55α, and Aim 3 to delineate the role of the PR55/YAP axis in pancreatic cancer development and progression in the KPC spontaneous mouse model. The work is innovative, as it will illuminate a critical pathway that promotes PC initiation and progression through PR55/PP2A activated YAP signaling, which is novel to both the PP2A field and the PC field. Successful completion of the proposed studies will yield an in-depth mechanistic understanding of the critical contribution of PR55/PP2A complex to the observed cooperation between Kras activation and the loss of the p53 tumor suppressor during PC development and progression. The acquired knowledge not only will improve the understanding of the role of PP2A in cancer but also will contribute to identifying novel therapeutic targets for improving the treatment of PC.

摘要 胰腺癌(PC)是美国最致命的癌症之一，5年生存率为7%， 在过去的40年里基本上没有变化。尽管进行了广泛的研究，但没有有效的针对性 对这种疾病的治疗，表明对PC中的致癌信号网络了解不足。而当 几乎所有与癌症有关的信号网络都依赖于激酶和磷酸酶的协同作用 转导促进癌症的信号，大多数努力只放在癌症中的蛋白激酶的研究上 同时让蛋白质磷酸酶成为一个盲点。PP2A是一个杂三聚体家族，是主要的 人类细胞中丝氨酸/苏氨酸磷酸酶活性的来源，每个都由一个支架亚基(Aα/β)，一个 催化亚基(Cα/β)，以及控制底物的许多非保守的调节性(B)亚基之一(&gt；27 PP2A复合体的特异性。我们的初步研究发现，PP2A的一个调节亚基Pr55α是一种 促进恶性PC的重要新参与者。特别是，我们的研究表明，Pr55抑制了 河马肿瘤抑制通路中的MOB1/LATS自激活环，其功能是诱导 YAP的磷酸化导致其胞质滞留和蛋白酶体降解。符合基本原则 YAP信号在KRAS驱动的PC发生发展中的作用，敲除PR55α阻碍了 PC细胞的非锚定生长、致瘤性和转移。我们的初步数据还显示 抑癌基因P53及其靶基因对α蛋白稳定性的负性调节 FBXL20，是SCF(Skp1-CUL1-F-box蛋白)E3连接酶的底物接头。一贯地，我们观察到 Pr55α在PC标本中升高，其表达与患者的生存不良有关。基于这些 结果，我们假设P53功能的丧失会导致PR55α蛋白的稳定，这反过来又会导致 激活Kras所需的YAP致癌信号，从而驱动PC的启动和进展。我们将测试 三个特定目的的假说：目的1确定Pr55α/PP2A促进YAP的机制 目的2阐明该级联蛋白的激活及其在PC致癌和恶性潜能中的意义 P53是如何抑制pR55α水平的？目的3：阐明pR55/YAP轴在胰腺中的作用 KPC自发小鼠模型中肿瘤的发生和发展。这项工作是创新的，它将会 通过pR55/PP2A激活的YAP阐明促进PC启动和进展的关键途径 信令，这在PP2A领域和PC领域都是新的。圆满完成拟议的研究 将从机制上深入理解Pr55/PP2A复合体对 PC发生过程中Kras激活与P53抑癌基因缺失的协同作用 和进步。所获得的知识不仅将提高对PP2A在癌症中作用的理解 但也将有助于确定新的治疗靶点，以改善PC的治疗。

项目成果

The GSK3 kinase and LZTR1 protein regulate the stability of Ras family proteins and the proliferation of pancreatic cancer cells.

• DOI: 10.1016/j.neo.2022.01.002
• 发表时间: 2022-03
• 期刊: Neoplasia (New York, N.Y.)
• 作者: Palanivel C;Chaudhary N;Seshacharyulu P;Cox JL;Yan Y;Batra SK;Ouellette MM
• 通讯作者: Ouellette MM

p53/FBXL20 axis negatively regulates the protein stability of PR55α, a regulatory subunit of PP2A Ser/Thr phosphatase.

• DOI: 10.1016/j.neo.2021.10.002
• 发表时间: 2021-12
• 期刊: Neoplasia (New York, N.Y.)
• 作者: Madduri LSV;Brandquist ND;Palanivel C;Talmon GA;Baine MJ;Zhou S;Enke CA;Johnson KR;Ouellette MM;Yan Y
• 通讯作者: Yan Y

Benzimidazole carbamate induces cytotoxicity in breast cancer cells via two distinct cell death mechanisms.

• DOI: 10.1038/s41420-023-01454-6
• 发表时间: 2023-05-13
• 期刊: CELL DEATH DISCOVERY
• 影响因子: 7
• 作者: Graff, Brendan T.;Palanivel, Chitra;Jenkins, Christopher B.;Baranowska-Kortylewicz, Janina;Yan, Ying
• 通讯作者: Yan, Ying

Cell Signaling Pathways That Promote Radioresistance of Cancer Cells.

• DOI: 10.3390/diagnostics12030656
• 发表时间: 2022-03-08
• 期刊: Diagnostics (Basel, Switzerland)
• 作者: Ouellette MM;Zhou S;Yan Y
• 通讯作者: Yan Y