Inhibition of Cancer stem cell characterisitcs in Pancreatic Cancer

抑制胰腺癌中的癌症干细胞特征

• 批准号: 8334254
• 负责人: Sharmila Shankar
• 金额: --
• 依托单位: KANSAS CITY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-10-01 至 2016-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8334254
• 关键词: Apoptosis; Area; BCL2 gene; BIRC4 gene; Binding; Biological; CD44 gene; Cancer Patient; Cell surface; Cells; Characteristics; Chemopreventive Agent; Data; Development; Disease; Disease Progression; Drug resistance; Epithelial; Event; Failure; Goals; Growth; Human; Implant; In Vitro; Knowledge; Lesion; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Mesenchymal; Modeling; Molecular; Mus; Neoplasm Metastasis; Oral Administration; Outcome; Pancreas; Pancreatic Intraepithelial Neoplasia; Patients; Pharmaceutical Preparations; Play; Prevention; Prevention strategy; Preventive; Primary Neoplasm; Radiation therapy; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Pathway; Snails; Sonic Hedgehog Pathway; Staging; Stem cells; Sulforaphane; Testing; Therapeutic; Transgenic Mice; Undifferentiated; Up-Regulation; base; cancer initiation; cancer prevention; cancer stem cell; cancer therapy; carcinogenesis; cell behavior; cell growth; chemotherapy; chromatin immunoprecipitation; clinically significant; cruciferous vegetable; design; empowered; gemcitabine; improved; in vivo; lapatinib; migration; mouse model; notch protein; novel; novel strategies; pancreatic cancer cells; pancreatic neoplasm; pluripotency; precursor cell; prevent; prognostic; public health relevance; response; self-renewal; slug; small hairpin RNA; smoothened signaling pathway; stemness; tumor; tumor growth; tumor progression; tumorigenic

DESCRIPTION (provided by applicant): ABSTRACT Cancer stem cells (CSCs) have been proposed recently to be the cause cancer initiation, progression and chemotherapy failure in pancreatic cancer. Abberant hedgehog signaling occurs in pancreatic cancer during the progression of PanIN lesions to metastatic tumors, suggesting that sonic hedgehog (Shh) signaling may be an early event leading to accumulation of undifferentiated precursor cells in pancreatic cancer. This project is based on the premise that sulforaphane (SFN), a natural compound from the cruciferous vegetables, can be used for prevention and/or treatment of human pancreatic cancer. Unfortunately, the intracellular mechanisms by which SFN inhibits growth and metastasis, and induces apoptosis in pancreatic cancer cells and CSCs have never been examined. Thus, the main objective of this application is to examine the molecular mechanisms by which sulforaphane inhibits pancreatic CSC characteristics, and assess the chemopreventive / therapeutic potential of sulforaphane against pancreatic cancer by targeting CSCs. These studies will empower us with new knowledge that will guide us in formulating strategies for prevention of human pancreatic cancer by targeting CSCs. We have recently demonstrated that human CD133+CD44+CD24+ESA+ CSCs express Nanog, Notch-1 and Oct-4 and are highly tumorigenic in mice. Our preliminary data indicate that: (i) SFN inhibits self-renewal capacity of pancreatic CSCs; (ii) SFN inhibits the expression of pluripotency maintaining factors (Nanog, Oct-4 and Sox-2), epithelial mesenchymal (EMT) markers (Zeb-1, Snail, Slug), Bcl-2 and XIAP in pancreatic CSCs; (iii) SFN inhibits the expression of Smo, Ptch as well as effector molecule Gli 1 and 2, suggesting the clinical significance of Shh pathway in pancreatic cancer; (iv) enforced activation of Shh pathway or inhibition of Gli1 plus Gli2 expression by shRNA blocked the inhibitory effects of SFN, suggesting these effects of sulforaphane are mediated through Shh pathway; and (v) SFN inhibits self-renewal capacity of CSCs (CD133+CD44+CD24+ESA+ cells with high Nanog) isolated from pancreas of human primary tumors and KrasG12D mice. Based on our preliminary findings, we hypothesize that sulforaphane alone or in combination with standard chemotherapeutic drugs inhibit pancreatic cancer stem cell characteristics by regulating pluripotency promoting factors and inhibiting the sonic hedgehog pathway. The above hypothesis will be tested by the following four specific aims: Aim 1. To examine the molecular mechanisms by which sulforaphane inhibits human pancreatic cancer stem cell characteristics and sensitizes drug- resistant cancer stem cells in vitro. Aim 2. To examine whether sulforaphane inhibits self-renewal capacity, migration, invasion and epithelial mesenchymal transition of pancreatic cancer stem cells through sonic hedgehog pathway and further to examine the involvement of Shh pathway in tumor-stroma interaction. Aim 3. To examine the effects of oral administration of sulforaphane on pancreatic carcinogenesis using a KrasG12D transgenic mouse model. Aim 4. To validate whether oral administration of sulforaphane is effective in inhibiting pancreatic cancer stem cell characteristics and tumor growth by inhibiting sonic hedgehog pathway, and to examine the interactive effects of sulforaphane with standard chemotherapeutic drugs in a NOD/SCID/IL2Rgammanull mouse model. Our studies are very novel and significant because they have prognostic relevance and inhibit pancreatic cancer initiation and progression by targeting CSCs.

描述（由申请人提供）： 摘要：癌症干细胞（CSC）最近被认为是胰腺癌癌症发生、进展和化疗失败的原因。在 PanIN 病变进展为转移性肿瘤的过程中，胰腺癌中出现了异常的刺猬信号传导，这表明声波刺猬 (Shh) 信号传导可能是导致胰腺癌中未分化前体细胞积累的早期事件。该项目的前提是萝卜硫素（SFN）是一种来自十字花科蔬菜的天然化合物，可用于预防和/或治疗人类胰腺癌。不幸的是，SFN 抑制胰腺癌细胞和 CSC 生长和转移以及诱导细胞凋亡的细胞内机制尚未得到研究。因此，本申请的主要目的是研究萝卜硫素抑制胰腺CSC特性的分子机制，并评估萝卜硫素通过靶向CSC对胰腺癌的化学预防/治疗潜力。这些研究将为我们提供新知识，指导我们制定针对 CSC 预防人类胰腺癌的策略。我们最近证明人类 CD133+CD44+CD24+ESA+ CSC 表达 Nanog、Notch-1 和 Oct-4，并且在小鼠中具有高度致瘤性。我们的初步数据表明：（i）SFN 抑制胰腺 CSC 的自我更新能力； (ii) SFN 抑制胰腺 CSC 中多能性维持因子（Nanog、Oct-4 和 Sox-2）、上皮间质 (EMT) 标记物（Zeb-1、Snail、Slug）、Bcl-2 和 XIAP 的表达； (iii) SFN抑制Smo、Ptch以及效应分子Gli 1和2的表达，提示Shh通路在胰腺癌中的临床意义； (iv) 强制激活Shh通路或通过shRNA抑制Gli1和Gli2表达阻断了SFN的抑制作用，表明萝卜硫素的这些作用是通过Shh通路介导的； (v) SFN 抑制从人类原发性肿瘤和 KrasG12D 小鼠胰腺中分离的 CSC（具有高 Nanog 的 CD133+CD44+CD24+ESA+ 细胞）的自我更新能力。根据我们的初步研究结果，我们假设萝卜硫素单独或与标准化疗药物联合使用，通过调节多能性促进因子和抑制声波刺猬通路来抑制胰腺癌干细胞特征。上述假设将通过以下四个具体目标进行检验： 目标1. 探讨萝卜硫素在体外抑制人胰腺癌干细胞特性并使耐药肿瘤干细胞敏感的分子机制。目的2.探讨萝卜硫素是否通过Sonic Hedgehog通路抑制胰腺癌干细胞的自我更新能力、迁移、侵袭和上皮间质转化，并进一步探讨Shh通路在肿瘤间质相互作用中的参与。目标 3. 使用 KrasG12D 转基因小鼠模型检查口服萝卜硫素对胰腺癌发生的影响。目的 4. 在 NOD/SCID/IL2Rgammanull 小鼠模型中验证口服萝卜硫素是否能有效通过抑制 sonic hedgehog 通路来抑制胰腺癌干细胞特征和肿瘤生长，并检查萝卜硫素与标准化疗药物的相互作用。我们的研究非常新颖且意义重大，因为它们具有预后相关性，并通过靶向 CSC 来抑制胰腺癌的发生和进展。