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: 8597933
• 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/8597933
• 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)： 摘要近年来，肿瘤干细胞被认为是胰腺癌发生、发展和化疗失败的原因。在胰腺癌从Panin病变发展到转移肿瘤的过程中，胰腺癌中出现了异常的Hedgehog信号，提示Sonic Hedgehog(Shh)信号可能是导致胰腺癌未分化前体细胞聚集的早期事件。该项目的前提是萝卜硫素(SFN)，一种从十字花科蔬菜中提取的天然化合物，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CSCs生长和转移以及诱导细胞凋亡的细胞内机制从未被研究过。因此，本应用的主要目的是研究萝卜硫素抑制胰腺CSC特性的分子机制，并通过靶向CSCs来评估萝卜硫素对胰腺癌的化学预防/治疗潜力。这些研究将赋予我们新的知识，指导我们通过靶向CSCs来制定预防人类胰腺癌的策略。我们最近证实，人CD133+CD44+CD24+ESA+CSCs表达Nanog、Notch-1和Oct-4，并在小鼠中具有高致瘤性。我们的初步研究结果表明：(1)SFN抑制胰腺CSCs的自我更新能力；(2)SFN抑制胰腺CSCs中多能维持因子(Nanog、Oct-4和SOX-2)、上皮间充质(EMT)标志物(Zeb-1、Snail、Slug)、Bcl2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；(4)shRNA激活Shh途径或抑制Gli1+Gli2的表达可阻断SFN的抑制作用，提示萝卜硫素的这种作用是通过Shh途径介导的；(V)SFN抑制人原发肿瘤和KrasG12D小鼠胰腺CSCs(CD133+CD44+CD24+ESA+高Nanog细胞)的自我更新能力。根据我们的初步发现，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Sonic Hedgehog途径来抑制胰腺癌干细胞的特性。上述假说将通过以下四个具体目标进行验证：目的1.检测萝卜硫素在体外抑制人胰腺癌干细胞特性和增敏耐药肿瘤干细胞的分子机制。目的2.检测萝卜硫素是否通过Sonic Hedgehog途径抑制胰腺癌干细胞的自我更新能力、迁移、侵袭和上皮间充质转化，并进一步探讨Shh通路在肿瘤-间质相互作用中的作用。目的：利用KrasG12D转基因小鼠模型，观察口服萝卜硫素对胰腺癌发生的影响。目的4.在NOD/SCID/IL2RGammanull小鼠模型上，验证萝卜硫素能否通过抑制Sonic Hedgehog途径有效地抑制胰腺癌干细胞特性和肿瘤生长，并观察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和有意义，因为它们与预后相关，并通过靶向CSCs来抑制胰腺癌的发生和发展。