Characterization of NYP Peptides in Prostate Cancer

NYP 肽在前列腺癌中的表征

• 批准号: 9901482
• 负责人: Everardo Macias
• 金额: $ 21.01万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9901482
• 关键词: Adipocytes; Affinity; Aftercare; Agonist; Amino Acids; Antigens; Binding; Biogenesis; Biological; Biological Assay; Biological Markers; Biological Process; CWR22Rv1; Cancerous; Cell Line; Cellular biology; Cessation of life; Cleaved cell; Clinical; Development; Dipeptidyl-Peptidase IV; Doxycycline; Event; Exhibits; Feeding behaviors; G-Protein-Coupled Receptors; Genes; Genetic Screening; Genomics; Gleason Grade for Prostate Cancer; Growth; Human; Hypothalamic structure; Immunohistochemistry; In Vitro; Kinetics; Kininogenase; Knock-in Mouse; Length; Libraries; Ligands; Link; Male Genital Organs; Malignant Neoplasms; Malignant neoplasm of prostate; Mass Spectrum Analysis; Membrane; Mus; Neoplasm Metastasis; Neurons; Neuropeptide Y Receptor; Neurotransmitters; Obese Mice; Obesity; Open Reading Frames; PSA screening; Pathway interactions; Peptide Fragments; Peptide Hydrolases; Peptides; Peptidyl-Dipeptidase A; Periprostatic; Phenotype; Phosphotransferases; Physiology; Plasma; Plasma Kallikrein; Play; Property; Prostate; Prostate-Specific Antigen; Prostatic; Protease Inhibitor; Protein-Serine-Threonine Kinases; Radical Prostatectomy; Reading Frames; Reporter; Reporting; Ribosomes; Role; Serine; Serine Protease; Serum; Signal Transduction; Specificity; Specimen; Stable Isotope Labeling; Testing; Tetracyclines; Tissues; Transgenic Mice; Uncertainty; cancer biomarkers; cancer recurrence; differential expression; drug metabolism; high risk; human tissue; in vivo; knock-down; matrigel; nerve supply; neuropeptide Y; neuropeptide Y2 receptor; novel; overexpression; patient response; promoter; prostate cancer cell; prostate cancer cell line; prostate carcinogenesis; receptor; small hairpin RNA; steroid hormone biosynthesis; transcriptome sequencing; tumor growth; tumorigenesis; tumorigenic

ABSTRACT Obesity is linked with greater risk of high-grade prostate cancer (PC), recurrence after therapy, metastases, and PC death. We exploited the link between obesity and aggressive prostate to identify actionable targets. To this end, we performed a shRNA genomic screen in obese mice targeting the entire kinome. Our screen identified Right Open Reading Frame Kinase 2 (RIOK2), as a promising PC target in obese mice and suggests that ribosomal biogenesis plays a role in PC. Our subsequent studies have identified non-ribosomal RIOK2 functions that link Neuropeptide Y (NPY), a neurotransmitter involved in feeding behavior and obesity, with PC cell biology. We found RIOK2 regulates Neuropeptide Y2 Receptor (NYP2R), one of five known mammalian neuropeptide Y receptors designated Y1 through Y5, which bind to NPY. Mechanistically, RIOK2 binds the NPY2R promoter and depletion of RIOK2 significantly reduces NPY2R expression. We found knockdown of NPY2R in PC cells, inhibits proliferation and reduces tumorigenic properties of PC cells in vitro. NPY, the NPY2R ligand, is chiefly produced in hypothalmic neurons, yet the male genital track also expresses NPY secreted from innervation, including in prostate and periprostatic adipocytes. NPY, is further upregulated in PC (~50 fold). In fact, NPY is a reported PC biomarker. Yet, it is unclear if this neurotransmitter plays a biological role in PC development or progression. Adding to the uncertainty, the full length 36 amino acid NPY1–36 is readily cleaved to NPY3–36, making it a selective NPY2R agonist by a dipeptidyl peptidase IV (DPP4), a membrane protease is also highly expressed in PC. Thus, NPY3-36 likely accounts for a large portion of NPY peptides in PC, which may elicit distinct signaling events compared to full length NPY1-36 primarily via NPY2R. However, prostate serum antigen aka kallikerin 3, a prostate serum biomarker is also a peptidase that may act on NPY. Our overarching hypothesis is that NPY is readily cleaved by prostatic peptidases and NPY peptide fragments play a role in PC. Mechanistically, we hypothesize that NPY is cleaved by prostatic DPP4 and/or PSA, and prostatic NPY cleavage peptides have differential downstream signaling cascades and phenotypic effects compared to full length NPY. To test this hypothesis we will (Aim 1) identify and characterize prostatic NPY cleaved peptides and (Aim 2) test if NPY levels alter PC development in vivo and in ex vivo human PCs.

抽象的 肥胖与高级前列腺癌（PC）的风险更大，治疗后复发，转移和 PC死亡。我们探索了肥胖和侵略性前列腺之间的联系，以识别可起诉的靶标。对此 最后，我们在靶向整个Kinome的肥胖小鼠中进行了shRNA基因组筛查。我们的屏幕确定了 右开放式阅读框架2（RIOK2），作为肥胖小鼠的承诺PC目标，并暗示 核糖体生物发生在PC中起作用。我们随后的研究确定了非核糖体RIOK2功能 该神经肽Y（NPY）是一种与PC细胞生物学有关的神经递质，涉及喂养行为和肥胖的神经递质。 我们发现RIOK2调节神经肽Y2受体（NYP2R），这是五个已知的哺乳动物神经肽Y之一 接收器指定Y1至Y5，与NPY结合。从机械上讲，RIOK2结合NPY2R启动子 RIOK2的耗竭显着降低了NPY2R的表达。我们发现PC细胞中NPY2R的敲低， 抑制体外PC细胞的肿瘤性特性，抑制增殖和降低。 NPY，NPY2R配体主要是 在假设神经元中产生，但男性生殖器轨迹也表达了从神经上分泌的NPY， 包括前列腺和腹膜脂肪细胞。 NPY在PC中进一步更新（〜50倍）。实际上，npy是一个 报告了PC生物标志物。但是，尚不清楚该神经递质在PC开发中起生物学作用还是 进展。除了不确定性外，全长36氨基酸NPY1-36很容易裂解至NPY3-36，使 它是二肽基肽IV（DPP4）的选择性NPY2R激动剂，膜蛋白酶也高度表达 在PC中。这是NPY3-36可能是PC中的大部分NPY Pepperides，这可能引起不同的信号 与全长NPY1-36主要通过NPY2R相比。但是，前列腺血清抗原又名kallikerin 3， 前列腺血清生物标志物也是一种可能作用于NPY的肽。我们的总体假设是NPY 很容易被前列腺宠物酶和NPY胡椒片段裂解在PC中发挥作用。机械上， 我们假设NPY被前列腺DPP4和/或PSA裂解，并且前列腺NPY裂解肽具有 与全长NPY相比，差异下游信号级联和表型效应。测试这个 假设我们将（目标1）识别和表征前列腺NPY裂解的Petides，并且（AIM 2）测试NPY如果 水平会改变体内和体内人体PC中的PC发育。