Developing glycosylated RNAs as novel clinical targets for aggressive prostate cancer.

开发糖基化 RNA 作为侵袭性前列腺癌的新临床靶点。

基本信息

批准号：
10651206
负责人：
Aurora Esquela Kerscher
金额：
$ 21.04万
依托单位：
EASTERN VIRGINIA MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10651206
关键词：
Animals Apoptosis Area Azides Benign Biological Biological Markers Biological Process Biotinylation Cancer Biology Cancer Etiology Cancer Patient Carbohydrates Cell Line Cell surface Cells Cellular Membrane Cessation of life Chemicals Chemistry Classification Clinical Colon Comparative Study Cultured Cells Diagnosis Disease Disease Progression Distant Ensure Enzymes Extracellular Matrix Fractionation Glycolipids Glycoproteins Golgi Apparatus Health Homeostasis Hormones Human Immune System Diseases Immune response Immunologic Surveillance Inosine Invaded Knockout Mice Label Lectin Link Lipids Liquid substance Malignant - descriptor Malignant Neoplasms Malignant neoplasm of prostate Mass Spectrum Analysis Mediating Metabolic Methods Modification Molecular Monitor Morphologic artifacts Mus Mutant Strains Mice Neoplasm Metastasis Non-Malignant Northern Blotting Nucleic Acids Oncogenic Organ Pancreas Pathology Pathway interactions Patient-Focused Outcomes Patients Pattern Pharmaceutical Preparations Play Polysaccharides Population Post-Transcriptional RNA Processing Post-Translational Protein Processing Process Prognosis Prostate Prostate Cancer therapy Prostatic Intraepithelial Neoplasias Prostatic Neoplasms Protein Glycosylation Proteins Pseudouridine RNA Recurrent disease Role Screening procedure Sialic Acids Signal Transduction Site Survival Rate Tail Testing Therapeutic Time Tissues Tumor Suppressor Genes United States Untranslated RNA Uridine Work advanced disease angiogenesis biomarker development cancer biomarkers cancer prevention cancer therapy cell growth curative treatments exosome extracellular vesicles glycosylation hormone sensitivity human disease improved in vivo inhibitor innovation insight intercellular communication male men mouse model novel nucleocytoplasmic transport patient stratification posttranscriptional prostate cancer cell prostate cancer cell line prostate cancer model prostate cancer progression response screening sialylation sugar survivorship tool transcriptome sequencing translational potential treatment strategy tumor growth tumor progression tumorigenic virtual

项目摘要

PROJECT SUMMARY This exploratory proposal will be the first to characterize glycosylated RNAs in the context of human prostate cancer (PCa). PCa is the most prevalent form of non-skin cancer in males and second leading cause of cancer-related deaths among men. 20% of men diagnosed with PCa will progress to fast-growing, advanced disease. There are no curative treatments for PCa that has spread to distant sites and the 5-year survival rate for metastatic disease is only 30%. As high-grade PCa often leads to poor prognoses, it is imperative to better understand these pathologies and improve patient stratification methods as well as therapeutic treatment options to increase patient survival. Noncoding RNAs are widely misexpressed in PCa patients and act as key tumor suppressor genes, pro-oncogenic factors in the prostate. Noncoding RNAs harbor a large range of post- transcriptional modifications (m6A, inosine, pseudouridine, 2'-O-Me, A-to-I editing, 3’ uridine tailing) that are important for RNA maturation, folding, expression, and nuclear transport. Dysregulation of these post- transcriptional modifications are associated with tumor growth, invasion, angiogenesis, immune response and disease recurrence. Noncoding RNAs were serendipitously discovered to carry glycosylation modifications when cultured human and mouse cells were metabolically labeled using bioorthogonal chemistry methods normally employed for glycosylated protein and lipid enrichment. Glycosylation is an intricate process that typically involves the covalent attachment of carbohydrates onto proteins and lipids as the biomolecules move through the secretory pathway. Aberrant protein/lipid glycosylation contributes to tumor growth, metastasis, and immunosurveillance evasion and is being utilized as cancer biomarkers. We hypothesize that RNAs require glycosylation for cell signaling to maintain prostate homeostasis and cancer prevention and therefore the glycosylated state of RNA will correlate with prostate tumorgenicity. In support of this rationale, we confirmed using click chemistry and northern blotting that glycoRNAs exist and with differing abundance in human prostate cells. Specific Aims: This proposal will use two independent methods, metabolic labeling with azide click chemistry and lectin-based purification, to classify glycoRNA distribution from a panel of human prostate cell lines differing in their metastatic potential and hormone sensitivity. Aim 1 will unbiasedly determine if small versus large noncoding RNAs are preferentially glycosylated in the prostate, if these glycoRNAs share features for N- or O-linked sugars, and identify these species using RNAseq. In Aim 2, fractionation methods and chemical inhibitor studies will determine the PCa-associated glycoRNA subcellular localization, possible exosome enrichment and characterize these carbohydrate moieties via mass spectrometry. In Aim 3, the Ptenpc-/- Smad4 pc-/- double knockout mouse model will be employed to characterize glycoRNAs from living animals throughout a prostate cancer progression time course. This work will lead to novel insights into how RNA modifications impact PCa progression and identify first-in-class clinical tools to improve patient outcome.

项目摘要该探索性建议将是第一个在人类前列腺中表征糖基化RNA 癌症（PCA）。 PCA是男性中最普遍的非皮肤癌的形式，第二个主要原因是男性与癌症有关的死亡。 20％被诊断为PCA的男性将发展为快速增长的高级疾病。没有针对PCA的治疗方法扩展到远处和5年生存率对于转移性疾病，只有30％。由于高级PCA通常会导致预后不佳，因此必须更好了解这些病理并改善患者分层方法以及治疗治疗增加患者生存的选择。未编码的RNA在PCA患者中被广泛采用，并充当关键肿瘤抑制基因，前列腺中促源性因子。未编码的RNA携带大量的后转录修改（M6A，inosine，pseudouridine，2'-o-me，a-to-i编辑，3'尿苷尾巴）对于RNA成熟，折叠，表达和核转运至关重要。这些后这些失调转录修饰与肿瘤的生长，侵袭，血管生成，免疫响应和疾病复发。偶然发现非编码RNA以进行糖基化修饰使用生物正交化学方法将培养的人类和小鼠细胞代谢标记通常用于糖基化蛋白质和脂质富集。糖基化是一个复杂的过程通常，随着生物分子的移动穿过秘密路径。异常蛋白/脂质糖基化有助于肿瘤生长，转移，和免疫监视的演变，并被用作癌症生物标志物。我们假设RNA 需要糖基化以维持前列腺稳态和预防癌症，因此 RNA的糖基化状态将与前列腺肿瘤性相关。为了支持这个理由，我们使用点击化学和北印迹确认糖原存在，并且有不同的丰度人前列腺细胞。具体目的：该提案将使用两种独立方法，包括代谢标签叠氮化物点击化学和基于讲座的纯化，以对人类小组进行分类前列腺细胞系的转移潜力和骑马敏感性不同。 AIM 1将公正地确定如果在前列腺中优选的小型非编码RNA是糖基化的，则这些糖菌共享 N-或O连接糖的特征，并使用RNASEQ识别这些物种。在AIM 2中，分馏方法化学抑制剂研究将确定与PCA相关的糖性亚细胞定位，可能外泌体富集并通过质谱法表征这些碳水化合物部分。在AIM 3中 PTENPC - / - SMAD4 PC - / - 双基因敲除鼠标模型将被雇用以表征Live的Glycornas 整个前列腺癌进展时间过程中的动物。这项工作将导致对如何 RNA修饰会影响PCA的进展并确定一流的临床工具以改善患者预后。