Aspartate β-hydroxylase promotes pancreatic ductal adenocarcinoma metastasis through activation of SRC signaling pathway
Aspartate β-hydroxylase promotes pancreatic ductal adenocarcinoma metastasis through activation of SRC signaling pathway
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DOI：
10.1186/s13045-019-0837-z
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发表时间：
2019-12-30
影响因子：
28.5
通讯作者：
Dong, Xiaoqun
中科院分区：
文献类型：
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作者：
Ogawa, Kosuke;Lin, Qiushi;Dong, Xiaoqun
背景：在成人胰腺肿瘤发生过程中，对胚胎发育至关重要的信号通路再次出现。天冬氨酸β -羟化酶（ASPH）在胰腺发育/分化过程中驱动胚胎细胞的运动/侵袭。我们探讨了失调的ASPH是否在胰腺癌发病机制中起关键作用。方法：为了证明ASPH是否/如何介导恶性表型、增殖、迁移、2-D/3-D侵袭、胰球形成、免疫荧光、Western blot、共免疫沉淀、侵入性形成/成熟/功能、qRT-PCR、免疫组织化学（IHC）和自行开发的体外转移试验。建立了人胰腺导管腺癌（PDAC）患者源异种移植（PDX）模型，以说明第三代小分子抑制剂特异性抗ASPH β -羟化酶活性的体内抗肿瘤作用。应用Kaplan-Meier图、log-rank检验和Cox比例风险回归模型评估ASPH网络成分的预后价值。结果：ASPH使胰腺癌细胞具有更强的侵袭性表型，其特征是上皮-间质转化（EMT）， 2-D/3-D侵袭，浸润形成/功能，如细胞外基质（ECM）降解，干性（癌症干细胞标记物升高和胰腺圈形成），跨内皮迁移（模拟内渗/外渗）和球体形成（模拟远处转移性定植/生长）。在机制上，ASPH通过与ADAM12/ADAM15的直接物理相互作用激活SRC级联，而不依赖于FAK。ASPH-SRC轴促进侵过性构建，并启动mmp介导的ECM降解/重塑，作为侵袭性的执行者。invadopdia的药理学抑制可减弱体外转移。在PDAC的PDX模型中，ASPH促进原发肿瘤的发展和肺转移，这是由一种专门针对ASPH酶活性的先导化合物阻断的。在正常胰腺中，ASPH是沉默的，从恶性病变前到PDAC侵袭性/晚期，ASPH逐渐上调。ASPH-SRC网络组分的表达谱独立/联合预测PDAC患者的临床预后。与负低水平相比，中度至极高水平的ASPH、ADAM12、活化SRC和MMPs与胰腺癌患者总生存期（OS）缩短相关（log-rank检验，ps < 0.001）。患者携带的不良分子越多，预后就越差。5种分子0-2分（n = 4）、3-5分（n = 8）、6-8分（n = 24）、9-12分（n = 73）不良表达评分患者的中位生存时间分别为55.4、15.9、9.7、5.0个月（p < 0.001）。结论：靶向PDAC多步转移过程中至关重要的ASPH-SRC轴可能特异性/实质性地延缓PDAC的发生/进展，从而改善PDAC的预后。
Background: Signaling pathways critical for embryonic development re-emerge in adult pancreas during tumorigenesis. Aspartate beta-hydroxylase (ASPH) drives embryonic cell motility/invasion in pancreatic development/differentiation. We explored if dysregulated ASPH is critically involved in pancreatic cancer pathogenesis.Methods: To demonstrate if/how ASPH mediates malignant phenotypes, proliferation, migration, 2-D/3-D invasion, pancreatosphere formation, immunofluorescence, Western blot, co-immunoprecipitation, invadopodia formation/maturation/function, qRT-PCR, immunohistochemistry (IHC), and self-developed in vitro metastasis assays were performed. Patient-derived xenograft (PDX) models of human pancreatic ductal adenocarcinoma (PDAC) were established to illustrate in vivo antitumor effects of the third-generation small molecule inhibitor specifically against ASPH's beta-hydroxylase activity. Prognostic values of ASPH network components were evaluated with Kaplan-Meier plots, log-rank tests, and Cox proportional hazards regression models.Results: ASPH renders pancreatic cancer cells more aggressive phenotypes characterized by epithelial-mesenchymal transition (EMT), 2-D/3-D invasion, invadopodia formation/function as demonstrated by extracellular matrix (ECM) degradation, stemness (cancer stem cell marker upregulation and pancreatosphere formation), transendothelial migration (mimicking intravasation/extravasation), and sphere formation (mimicking metastatic colonization/outgrowth at distant sites). Mechanistically, ASPH activates SRC cascade through direct physical interaction with ADAM12/ADAM15 independent of FAK. The ASPH-SRC axis enables invadopodia construction and initiates MMP-mediated ECM degradation/remodeling as executors for invasiveness. Pharmacologic inhibition of invadopodia attenuates in vitro metastasis. ASPH fosters primary tumor development and pulmonary metastasis in PDX models of PDAC, which is blocked by a leading compound specifically against ASPH enzymatic activity. ASPH is silenced in normal pancreas, progressively upregulated from pre-malignant lesions to invasive/advanced stages of PDAC. Expression profiling of ASPH-SRC network components independently/jointly predicts clinical outcome of PDAC patients. Compared to a negative-low level, a moderate-very high level of ASPH, ADAM12, activated SRC, and MMPs correlated with curtailed overall survival (OS) of pancreatic cancer patients (log-rank test, ps < 0.001). The more unfavorable molecules patients carry, the more deleterious prognosis is destinated. Patients with 0-2 (n = 4), 3-5 (n = 8), 6-8 (n = 24), and 9-12 (n = 73) unfavorable expression scores of the 5 molecules had median survival time of 55.4, 15.9, 9.7, and 5.0 months, respectively (p < 0.001).Conclusion: Targeting the ASPH-SRC axis, which is essential for propagating multi-step PDAC metastasis, may specifically/substantially retard development/progression and thus improve prognosis of PDAC.