Sialylation-dependent mechanisms driving pancreatic cancer progression

唾液酸化依赖机制驱动胰腺癌进展

• 批准号: 10468125
• 负责人: Susan L Bellis
• 金额: $ 50.37万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-18 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10468125
• 关键词: Address; Apoptosis; Apoptotic; Area; Automobile Driving; Biochemical; Biological Assay; Biology; Cell model; Cells; Cessation of life; Characteristics; Colon; Competence; Data; Development; Disease; Duct (organ) structure; Enzymes; Epidermal Growth Factor Receptor; Epithelial; Event; Fostering; Genetic Transcription; Genetically Engineered Mouse; Glycobiology; Growth; Human; Implant; Injections; Integrins; KRAS2 gene; Ligation; Link; Literature; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Metaplasia; Metaplastic Cell; Metastatic Neoplasm to the Liver; Modeling; Modification; Molecular; Molecular Biology; Monitor; Mus; Neoplasm Metastasis; Oncogenic; Organoids; Pancreas; Pancreatic Adenocarcinoma; Pancreatic Ductal Adenocarcinoma; Pathogenesis; Patients; Phenotype; Play; Polysaccharides; Prognosis; Property; Resistance; Role; ST6Gal I; Seminal; Sialic Acids; Sialyltransferases; Signal Transduction; Stains; Surface; TNFRSF1A gene; Testing; Tissues; Tumor-Derived; Up-Regulation; Variant; Work; anticancer research; bioluminescence imaging; cancer stem cell; cell behavior; cohort; effective therapy; exosome; glycoproteomics; glycosylation; glycosyltransferase; in vivo; in vivo imaging system; mortality; mouse model; neoplastic cell; novel; overexpression; pancreatic cancer model; pancreatic ductal adenocarcinoma cell; receptor; sialylation; stem; stem-like cell; targeted treatment; transcription factor; tumor; tumor initiation; tumor progression; unpublished works

Summary Although it has been known for decades that the ST6Gal-I sialyltransferase is upregulated in cancer, there is still a striking dearth of information regarding the functional role of ST6Gal-I in regulating tumor cell behavior. Our group has been at the forefront of characterizing the specific ST6Gal-I receptor substrates that reprogram tumor cell signaling to impart a stem-like, metastatic phenotype. These studies implicate ST6Gal-I as a master regulatory molecule that controls the function (via sialylation) of key receptors including TNFR1, Fas, EGFR and β1 integrin. Together this receptor cohort directs the activation of signaling cascades that promote hallmark cancer stem cell (CSC) characteristics such as invasiveness and apoptosis-resistance. In recent unpublished work, we have definitively confirmed a tumor-driver role for ST6Gal-I in pancreatic ductal adenocarcinoma (PDAC). We generated a new genetically engineered mouse with ST6Gal-I overexpression in the pancreas, and crossed this line to the “KC” pancreatic cancer model, which harbors oncogenic K-ras expression in the pancreas. Compared to KC mice, KC mice with ST6Gal-I overexpression have greatly accelerated PDAC development, metastasis, and mortality. We hypothesize that ST6Gal-I activity contributes to two critical steps in pathogenesis: (1) acinar-to-ductal metaplasia (ADM), an early event in PDAC initiation (Aim 1), and (2) PDAC metastasis (Aim 2), which is the chief cause of patient mortality. In Aim 1 we will elucidate molecular mechanisms by which ST6Gal-I promotes ADM, focusing on the hypothesis that ST6Gal-I activates NF-ĸB signaling to induce Sox9 expression. A robust literature has established that upregulation of Sox9 plays an essential role in ADM, however the finding that ST6Gal-I-mediated sialylation controls Sox9 expression points to a transformative role for tumor glycosylation in PDAC initiation. The importance of a ST6Gal-I/NF-ĸB/Sox9 signaling axis in ADM will be evaluated in: (i) the canonical 266-6 ADM cell model; (ii) epithelial organoid lines derived from our various mouse models; and (iii) the in vivo ADM models, ductal ligation and cerulein injection. In Aim 2 we will interrogate the sialylation-dependent signaling mechanisms that reprogram tumor cells into CSC-like cells with metastatic capability. Our central premise is that ST6Gal-I- mediated sialylation of EGFR, TNFR1/Fas, and β1 integrin acts as a molecular switch to alter signaling nodes that confer stem-like properties. Furthermore, we will test the game-changing hypothesis that exosomal transfer of active ST6Gal-I to recipient cells can reprogram these cells to acquire metastatic properties. To verify that ST6Gal-I plays a causal role in metastasis we will use bioluminescence imaging to track the metastatic dissemination of orthotopically implanted PDAC patient organoids, as well as Suit2 cells and their isogenic metastatic clones. In the aggregate, these studies are expected to reveal an unprecedented role for tumor cell sialylation in driving PDAC progression.

总结 尽管几十年来已知ST 6 Gal-I唾液酸转移酶在癌症中上调，但仍存在一些缺陷。 关于ST 6 Gal-I在调节肿瘤细胞行为中的功能作用的信息仍然非常缺乏。 我们的团队一直处于表征重编程的特异性ST 6 Gal-I受体底物的最前沿。 肿瘤细胞信号传导以赋予干细胞样转移表型。这些研究表明，ST 6 Gal-I是一种主要的 控制关键受体（包括TNFR 1、Fas、EGFR）功能（通过唾液酸化）的调节分子 和β1整联蛋白。这种受体群共同指导信号级联的激活， 标志性癌症干细胞（CSC）特征，如侵袭性和抗肿瘤性。近几 在未发表的工作中，我们已经明确证实了ST 6 Gal-I在胰腺导管中的肿瘤驱动作用。 腺癌（PDAC）。我们产生了一种新的基因工程小鼠，其ST 6 Gal-I过表达于 胰腺，并越过这条线到达“KC”胰腺癌模型，该模型含有致癌K-ras 在胰腺中的表达。与KC小鼠相比，ST 6 Gal-I过表达的KC小鼠具有显著的 加速PDAC的发展、转移和死亡。我们假设ST 6 Gal-I活性有助于 发病机制的两个关键步骤：（1）腺泡-导管化生（ADM），PDAC启动的早期事件 (Aim 1），和（2）PDAC转移（目的2），这是患者死亡的主要原因。在目标1中， 阐明了ST 6 Gal-I促进ADM的分子机制，重点是ST 6 Gal-I促进ADM的假说， 激活NF-κ B信号传导以诱导Sox 9表达。一个强大的文献已经确定，上调 Sox 9在ADM中起重要作用，然而，发现ST 6 Gal-I介导的唾液酸化控制Sox 9， 表达指出肿瘤糖基化在PDAC起始中的转化作用。的重要性 ADM中的ST 6 Gal-I/NF-κ B/Sox 9信号传导轴将在以下中评估：（i）典型的266-6 ADM细胞模型;（ii） 来自我们的各种小鼠模型的上皮类器官系;和（iii）体内ADM模型，导管 结扎和蛙皮素注射。在目标2中，我们将询问唾液酸化依赖的信号传导机制， 将肿瘤细胞重编程为具有转移能力的CSC样细胞。我们的中心前提是，ST 6 Gal-I- EGFR、TNFR 1/Fas和β1整联蛋白介导的唾液酸化作为分子开关改变信号节点 赋予了茎状的特性。此外，我们将测试改变游戏规则的假设，即外泌体 将活性ST 6 Gal-I转移到受体细胞中可以重编程这些细胞以获得转移特性。到 为了证实ST 6 Gal-I在转移中起因果作用，我们将使用生物发光成像来跟踪ST 6 Gal-I在转移中的作用。 原位植入的PDAC患者类器官以及Suit 2细胞和其 同基因转移克隆。总的来说，这些研究预计将揭示一个前所未有的作用， 肿瘤细胞唾液酸化在驱动PDAC进展中的作用。

项目成果

Glycosyltransferase ST6Gal-I promotes the epithelial to mesenchymal transition in pancreatic cancer cells.

• DOI: 10.1074/jbc.ra120.014126
• 发表时间: 2021-01
• 期刊: The Journal of biological chemistry
• 作者: Britain CM;Bhalerao N;Silva AD;Chakraborty A;Buchsbaum DJ;Crowley MR;Crossman DK;Edwards YJK;Bellis SL
• 通讯作者: Bellis SL