Integrative Molecular Profiling of Human Pancreatic Cancer

该项目利用基于组学和靶向策略的组合来研究胰腺癌进展和疾病侵袭的潜在机制，以确定候选治疗靶点。在我们的调查过程中，与前面描述的第一个项目形成了几个交叉对话。在我们的一项研究中，我们试图获得对mif高表达和低表达肿瘤的全面分子洞察，并确定驱动肿瘤进展的关键mif信号通路，我们分析了PDAC患者多组中高（高于中位数）和低（低于中位数）mif表达肿瘤的编码和非编码基因的表达。所确定的关键基因和途径与患者的生存有关，并通过细胞系、基因工程小鼠模型和PDAC患者队列进行了机制、功能和临床表征。我们报道了一种新的mif驱动的信号通路的机制、功能和临床证据，该信号通路抑制先前未描述的潜在肿瘤抑制因子，核受体亚家族3，组c，成员2 (NR3C2)，导致PDAC的疾病侵袭性增强和生存率降低。在机制上，MIF上调miR-301b，然后靶向并抑制NR3C2的表达。MIF表达较高的肿瘤显示miR-301b升高，NR3C2表达降低。此外，在多个独立的PDAC患者队列中，肿瘤中NR3C2表达较低的患者生存率较低。这些发现发现了一个关键的mif诱导的信号通路，它可以促进PDAC的肿瘤进展和疾病侵袭性。该研究还发现NR3C2，一种矿化皮质激素受体基因，作为疾病进展的推定抑制剂。我们目前正在研究NR3C2在PDAC中的机制作用，涉及其调节代谢途径基因的潜在功能。此外，在我们最初的努力中，我们比较了早期PDAC患者的肿瘤基因表达谱，这些患者的生存期明显不同，早期PDAC患者的生存期极差（6个月）和手术切除后存活2年或更长时间。两组间1820个差异表达基因中炎症基因网络占主导地位。低表达的NOSTRIN与显著的低生存率相关，表明其在疾病进展中的潜在肿瘤抑制作用。NOSTRIN灭活内皮细胞NOS (eNOS/NOS3)，抑制一氧化氮（NO）的产生。此外，miR-221与NOSTRIN的3'UTR结合并抑制其表达，miR-221表达的增加与PDAC的生存率低相关。我们的研究结果与假设一致，即NOSTRIN是疾病侵袭性的潜在负调节因子，这可能是设计改进PDAC治疗策略的目标。我们正在研究no依赖性和no非依赖性在PDAC中的机制作用。如上所述，通过使用类似的策略来表征生存率明显不同的患者，我们进行了非靶向微rna分析，并鉴定了一种新的miR-331-3p/ADAMTSL3轴，该轴增强了切除胰腺癌患者的疾病侵袭性。如前所述，基于基因组、转录组学和代谢谱的差异，已经报道了胰腺导管腺癌的分子亚群。在这些亚群中具有治疗意义的特定分子靶点尚未明确定义。我们正在进行的研究正在调查一个高度侵袭性的亚组，以确定驱动肿瘤进展和疾病侵袭的关键机制，这些机制可能具有潜在的治疗意义。在我们的代谢组学研究中，我们已经表明，涉及脂肪酶和一组独特的游离脂肪酸的脂溶途径的损伤可能在胰腺癌的发生和进展中发挥重要作用，并为治疗干预提供潜在的靶点。我们目前正在研究炎症信号通路在胰腺癌代谢重编程调节中的作用。我们的工作还包括研究MIF-miR-301b-NR3C2信号轴在胰腺癌代谢重编程调节中的相互作用。此外，我们已经定义了一种新的NO/RUNX3/犬尿氨酸代谢信号通路，该通路增强了人类胰腺癌的疾病侵袭性，可能在改善疾病预后方面具有潜在的翻译意义。此外，我们正在对最具侵袭性的分子亚型进行深入分析，以了解疾病的侵袭性并确定潜在的治疗靶点。通过这种方法，我们研究了不同分子亚型癌症的代谢适应差异如何介导一种亚型与另一种亚型的疾病侵袭性。

This project utilizes a combination of omics-based and targeted strategies for investigating the underlying mechanisms of pancreatic cancer progression and disease aggressiveness to identify candidate therapeutic targets. During the course of our investigation several cross-talks have developed with the first project described earlier. In one of our studies, we tried to attain a comprehensive molecular insight into tumors with high- and low-MIF expression and to identify key MIF-signaling pathways that drive tumor progression, we analyzed the expression of coding and non-coding genes in high (above median) and low (below medianmedian) MIF-expressing tumors in multiple cohorts of PDAC patients. The identified key genes and pathways were linked to patients' survival and were mechanistically, functionally and clinically characterized using cell lines, genetically engineered mouse model and PDAC patient cohorts. We reported mechanistic, functional and clinical evidence of a novel MIF-driven signaling pathway that inhibits a previously undescribed potential tumor suppressor, nuclear-receptor-subfamily-3, group-C, member-2 (NR3C2), leading to enhanced disease aggressiveness and poorer survival in PDAC. Mechanistically, MIF upregulated miR-301b, which then targeted and suppressed NR3C2 expression. Tumors with a higher MIF expression showed an elevated miR-301b and a reduced NR3C2 expression. Additionally, patients with a lower NR3C2 expression in tumors showed poorer survival in multiple independent cohorts of PDAC patients. These findings discovered a key MIF-induced signaling pathway, which enhances tumor progression and disease aggressiveness in PDAC. This study also identified NR3C2, a mineralocorticoid receptor gene, as a putative inhibitor of disease progression. We are currently investigating the mechanistic role of NR3C2 in PDAC which involves it potential function in regulating the metabolic pathway genes. Furthermore, in our initial effort to characterize tumors from early stage resected PDAC patients with markedly different survival, we compared gene expression profiles in tumors from early stage PDAC cases with extremely poor survival (6 months) and those surviving 2 years or more following surgical resection. Inflammatory gene network dominated among 1,820 differentially expressed genes between the two groups. A lower expression of NOSTRIN was associated with significantly poor survival indicating its potential tumor inhibitory role in disease progression. NOSTRIN inactivated endothelial NOS (eNOS/NOS3) and inhibited the production of nitric oxide (NO). Furthermore, miR-221, bound to the 3'UTR of NOSTRIN and suppressed its expression, and an increased miR-221 expression associated with poor survival in PDAC. Our findings are consistent with the hypothesis that NOSTRIN is a potential negative regulator of disease aggressiveness, which may be targeted for designing improved treatment strategy in PDAC. We are investigating the NO-dependent and NO-independent mechanistic role of NOSTRIN in PDAC. By using a similar strategy to characterize the patients tumor with markedly different survival, as described above, we conducted an untargeted micro-RNA profiling and identified a novel miR-331-3p/ADAMTSL3 axis which enhances disease aggressiveness in resected pancreatic cancer patients As mentioned earlier, molecular subgroups of pancreatic ductal adenocarcinoma have been reported based on the differences in genomic, transcriptomics and metabolic profiling, however, specific molecular targets with therapeutic significance in these subgroups are yet to be clearly defined. Our ongoing study is investigating a highly aggressive subgroup to define key mechanisms driving tumor progression and disease aggressiveness that may be targeted with potential therapeutic significance. In our metabolomic studies, we have shown that impairment in a lipolytic pathway involving lipases and a unique set of free fatty acids, may play an important role in the development and progression of pancreatic cancer and provide potential targets for therapeutic intervention. We are currently investigating the role of inflammatory signaling pathways in the regulation of metabolic reprogramming in pancreatic cancer. Our effort also includes the investigation of an interactive role of MIF-miR-301b-NR3C2 signaling axis in the regulation of metabolic reprogramming in pancreatic cancer. Additionally, we have defined a novel NO/RUNX3/kynurenine metabolic signaling pathway which enhances disease aggressiveness in human pancreatic cancer and may have potential translational significance in improving disease outcome. Furthermore, we are conducting and in-depth analysis of the most aggressive molecular subtype to get an insight into the disease aggressiveness and identify potential therapeutic target. With this approach, we have investigated how the difference in metabolic adaptation in different molecular subtypes of cancer mediates the disease aggressiveness in one subtype versus another.