前列腺癌外泌体多基因预测模型及其侵袭性演变的分子机制

The application of prostate-specific antigen(PSA) in prostate cancer(PCa) screening has improved the positive diagnosis rate and the clinical management of PCa patients. Active surveillance for the indolent PCa and radical prostatectomy for the aggressive PCa were recommended. Thus, there is an urgent need for non-invasive PCa biomarkers with greater sensitivity and specificity that are capable of distinguishing aggressive disease from indolent disease during the active surveillance. Using whole genosome expression profile chip, we previously screened secretory genes including ERG, ITGB5, TIMP1 and TMEM176B which are differentially expressed between those patients with indolent and aggressive prostate cancer. The expression of these genes in peripheral blood and prostate cancer tissue were also confirmed using quantitative real-time PCR and IHC, respectively. Using weighted gene co-expression network analysis based on whole genosome expression profile chip, we will predict the genes on the pitch points and 4-gene regulating signal networks，which will validate the scientific hypothesis of the development of prostate cancer from indolent to aggressive stage. The present study aimed to establish the 4-gene detection in the exosome from the peripheral blood of prostate cancer patients. At the same time, the image texture features of multiparameter MRI are extracted and quantified to establish the radiomics method. This radiogenomics technique would be used as "liquid biopsy" for active surveillance or diagnosis of prostate cancer. The synergy/antagonism effect of 4-gene on prostate cancer invasion was further evaluated. we will further validate the genes on the pitch points and 4-gene mechanisms on regulating ERK, AKT, PKA and/or NF-κB signal networks. The conditional cell reprogramming technology will be used to establish the patient-derived cell bank of prostate cancer and verify the functional effects of four genes as a whole from indolent to aggressive PCa. Our study would evaluate the mechanism of 4-gene in driving prostate cancer progression, thus providing new clues for prostate cancer treatment strategy.

PSA筛查检出更多极早期前列腺癌(PCa)，惰性PCa主动监测和侵袭性PCa根治性治疗备受推荐，但缺乏有效的监测恶性演变指标。项目组前期利用基因芯片筛出惰性与侵袭性PCa组织间差异表达的四个分泌性基因(ERG,ITGB5,TIMP1,TMEM176B)，并验证其在PCa组织与外周血均差异表达。从生物信息学预测四基因最相关的节点基因(RPS11,RAP1GDS1)，提出PCa从惰性向浸润性性演变的科学假说。本项目拟提取PCa患者外周血外泌体RNA,定量检测四基因表达；提取并量化多参数MRI影像特征，建立影像基因组学预测模型，用于PCa诊断与主动监测的“液态活检”；研究四基因对PCa恶性演变的协同/拮抗作用以及如何调控ERK,AKT,PKA和/或NF-κB信号网络；拟利用条件性细胞重编程技术建立病人组织来源的细胞库，验证四基因作为整体在惰性向侵袭性PCa演变作用，阐明PCa恶性演变的分子机制。

前列腺癌的的诊断手段多样，但目前尚未出现一种无创的有效便捷的“液态活检”方法与生物学标记来监测前列腺癌的进程与恶性演变。建立一种方便有效的“液态活检”方法，用于鉴别和主动监测前列腺癌，并弄清其从惰性进展成侵袭性的分子机制，具有重大意义。本项目建立了四基因（ERG, ITGB5, TIMP1, TMEM176B）诊断前列腺癌以及鉴别惰性前列腺癌数学模型并通过分别构建四基因质粒载体，完成基因功能验证，阐明了对前列腺癌恶性演变的作用与调控机制。本项目采用条件重编程细胞技术在体外成功构建3例前列腺癌原代细胞株。本项目还收集了前列腺癌紫杉醇耐药细胞株分泌的外泌体，意在解释耐药性可以从紫杉醇耐药细胞株向紫杉醇敏感细胞株传递。本项目成功构建鉴别惰性前列腺癌和侵袭性前列腺癌的影像组学预测模型并对其进行验证本项目针对前列腺肿瘤微环境，设计了肿瘤微环境响应性自组装纳米粒，并在小鼠前列腺癌原位模型验证了其前列腺癌超声诊断与被动靶向治疗的能力。针对泌尿系肿瘤新型药物的开发，本项目开展了对CDK家族小分子抑制剂的研究，通过利用CRISPR-Cas9激酶组基因文库技术，筛选出CDK12作为前列腺癌有效的药物靶点，并进一步阐明了CDK12小分子抑制剂THZ531的抗癌作用机制和AR拮抗剂的协同效应。