Role of NF-kB signaling in supporting ovarian cancer tumor-initiating cells responsible for cancer recurrence

NF-kB 信号传导在支持卵巢癌肿瘤起始细胞（负责癌症复发）中的作用

• 批准号: 10046417
• 负责人: Carrie Danielle House
• 金额: $ 5.1万
• 依托单位: SAN DIEGO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10046417
• 关键词: Address; Adhesions; Award; Biology; Cancer Patient; Cancer Relapse; Cancer Remission; Cell Cycle; Cell Line; Cell Survival; Cell surface; Cells; Cellular biology; ChIP-seq; DNA Binding; Data; Development; Disease; Disease Resistance; Drug resistance; Family; Gene Expression Profiling; Gene Targeting; Genes; Genetic Transcription; Genomics; Goals; Immune response; In Vitro; Incidence; Individual; Inhibitor of Differentiation Proteins; Interruption; Investigation; MAP Kinase Gene; Malignant Female Reproductive System Neoplasm; Malignant neoplasm of ovary; Measures; Mediating; Molecular; Molecular Biology; Morbidity - disease rate; Mus; NF-kappa B; Outcome; Pathway interactions; Patients; Pharmacology; Phase; Phenotype; Platinum; Play; Population; Process; Proteins; Recurrence; Relapse; Reporter; Research; Research Design; Research Personnel; Resistance; Role; Secondary Prevention; Signal Pathway; Signal Transduction; Sorting - Cell Movement; System; Therapeutic; Training; Treatment Efficacy; Tumor stage; Tumorigenicity; United States; Woman; Xenograft Model; Xenograft procedure; base; cancer cell; cancer drug resistance; cancer recurrence; cancer type; cell type; chemotherapy; design; established cell line; in vivo; inhibitor/antagonist; insight; knock-down; monolayer; mortality; mouse model; neoplastic cell; novel; novel therapeutics; ovarian neoplasm; prevent; programs; public health relevance; small hairpin RNA; stem cells; tool; transcription factor; tumor; tumor initiation; tumorigenic

 DESCRIPTION (provided by applicant): The overall objective of this proposal is to define the role of the NF-kB pathway in ovarian cancer tumor-initiating cells (TICs) and to ascertain mechanisms required for successful repopulation of tumors following chemotherapy. Ovarian cancer is the most lethal gynecological malignancy in the United States and although most patients initially respond to platinum-based chemotherapy, over 70% of advanced stage tumors relapse leading to high morbidity and mortality. Thus, there is an urgent need to clarify processes that support therapy resistance and tumor regrowth, and the purpose of this research is to identify mechanisms that can be targeted to prevent tumor recurrence. NF-kB activity is associated with increased aggressiveness, enhanced metastatic potential and poor outcome in ovarian cancer. NF-kB is a ubiquitous signaling pathway whose target genes encode proteins that regulate immune response, cell survival, proliferation, adhesion, and interaction with the microenvironment. This proposal investigates the hypothesis that NF-kB pathways support subpopulations of ovarian cancer cells that can resist chemotherapy and drive tumor recurrence. The proposed project will address several questions relevant to the biology of ovarian cancer relapse. First, which functions of NF-kB support TICs that comprise a small percentage of the total cells in the tumor, and non-TICs that make up the bulk of the tumor? Are non- TICs necessary for efficient repopulation of the tumor? Does NF-kB activation integrate with MAPK/ERK to support non-TICs? Do transcriptional regulators that maintain stem cell phenotypes control NF-kB expression and activity? This proposal seeks to understand the coordination of multiple pathways in TICs and non-TICs necessary for ovarian cancer relapse. To begin answering these questions I first developed a culture system to enrich for TICs and compared NF-kB and MAPK/ERK activity in these cells with cells grown in non-TIC enriching conditions (adherent monolayer). My preliminary studies suggest classical NF-kB and MAPK/ERK activity support non-TICs whereas alternative NF-kB activity supports TICs. I will genetically interrupt the components leading to classical and alternative NF-kB activity in established cell lines and primary patient cell lines, and measure chemoresistance and tumor repopulation in vivo using xenograft mouse models. These studies will be extended in sorted TIC and non-TIC populations. Sorting will be accomplished using a novel reporter that responds to activity of stem cell transcription factors rather than traditional cell surface markers that ar context dependent. Investigation of pathways specifically activated in TICs or non-TICs will elucidate the potential cooperation of these different cell types in successful repopulation of tumors. Lastly, I will investigate novel mechanisms leading to alternative NF-kB activation in TICs to provide insights into new targets for prevention of secondary disease. During the K99 phase I will receive extensive training using mouse models to measure chemoresistance and tumor relapse and I will develop a reliable reporter for identifying and isolating ovarian cancer TICs. At the end of the K99 phase I will possess the tools necessary to be an independent investigator with expertise in xenograft mouse models and ovarian TIC biology. This new training will be applied to my overall study design during the R00 phase of this award, which aims to address significant gaps in our current understanding of ovarian cancer relapse. The expertise gained during the K99 phase, together with my molecular biology and genomics background, will allow me to develop an integrated research program designed to investigate novel mechanisms of ovarian cancer persistence and relapse, leading ultimately to new therapies to prolong the lives of women with this disease. Results of this proposal will advance the ovarian cancer field by providing mechanistic insight into ovarian cancer recurrence and the role of specific cell subtypes. Completion of the proposed aims will guide the development of alternative therapeutic strategies for patients with ovarian cancer and may highlight mechanisms relevant to other cancer types.

 描述（由申请人提供）：本提案的总体目标是确定NF-κ B通路在卵巢癌肿瘤起始细胞（TIC）中的作用，并确定化疗后肿瘤成功再增殖所需的机制。卵巢癌是美国最致命的妇科恶性肿瘤，尽管大多数患者最初对基于铂的化疗有反应，但超过70%的晚期肿瘤复发，导致高发病率和死亡率。因此，迫切需要澄清支持治疗抗性和肿瘤再生长的过程，本研究的目的是确定可以靶向预防肿瘤复发的机制。NF-kB活性与卵巢癌侵袭性增加、转移潜能增强和预后不良相关。NF-kB是一种普遍存在的信号通路，其靶基因编码调节免疫应答、细胞存活、增殖、粘附以及与微环境相互作用的蛋白质。该提案调查了NF-kB通路支持卵巢癌细胞亚群的假设，这些细胞亚群可以抵抗化疗并驱动肿瘤复发。拟议的项目将解决与卵巢癌复发的生物学相关的几个问题。首先，NF-kB的哪些功能支持占肿瘤细胞总数一小部分的TIC和占肿瘤大部分的非TIC？非TIC是肿瘤有效再增殖所必需的吗？NF-kB活化与MAPK/ERK整合以支持非TIC吗？维持干细胞表型的转录调节因子是否控制NF-κ B的表达和活性？该提案旨在了解卵巢癌复发所需的TIC和非TIC中多种途径的协调。为了开始回答这些问题，我首先开发了一种培养系统来富集TIC，并将这些细胞中的NF-κ B和MAPK/ERK活性与在非TIC富集条件下生长的细胞（贴壁单层）进行比较。我的初步研究表明，经典的NF-κ B和MAPK/ERK活性支持非TIC，而替代NF-κ B活性支持TIC。我将在已建立的细胞系和原代患者细胞系中遗传中断导致经典和替代NF-κ B活性的组分，并使用异种移植小鼠模型测量体内化疗耐药性和肿瘤再增殖。这些研究将在分类的TIC和非TIC人群中扩展。分选将使用响应干细胞转录因子的活性的新型报告基因而不是依赖于环境的传统细胞表面标记物来完成。对TIC或非TIC中特异性激活的通路的研究将阐明这些不同细胞类型在肿瘤成功再增殖中的潜在合作。最后，我将研究新的机制，导致替代NF-κ B激活的TIC提供新的目标，预防继发性疾病的见解。在K99阶段，我将接受广泛的培训，使用小鼠模型来测量化疗耐药性和肿瘤复发，我将开发一种可靠的报告基因，用于识别和分离卵巢癌TIC。在K99阶段I结束时，将拥有成为具有异种移植小鼠模型和卵巢TIC生物学专业知识的独立研究者所需的工具。这项新的培训将应用于我在该奖项的R 00阶段的整体研究设计，旨在解决我们目前对卵巢癌复发的理解存在的重大差距。在K99阶段获得的专业知识，加上我的分子生物学和基因组学背景，将使我能够开发一个综合研究计划，旨在研究卵巢癌持续性和复发的新机制，最终导致新的治疗方法，以延长患有这种疾病的妇女的生命。该提案的结果将通过提供对卵巢癌复发和特定细胞亚型的作用的机制见解来推进卵巢癌领域。完成拟议的目标将指导卵巢癌患者替代治疗策略的发展，并可能突出与其他癌症类型相关的机制。

项目成果

IΚΚε cooperates with either MEK or non-canonical NF-kB driving growth of triple-negative breast cancer cells in different contexts.

• DOI: 10.1186/s12885-018-4507-2
• 发表时间: 2018-05-25
• 期刊: BMC cancer
• 影响因子: 3.8
• 作者: House CD;Grajales V;Ozaki M;Jordan E;Wubneh H;Kimble DC;James JM;Kim MK;Annunziata CM
• 通讯作者: Annunziata CM

NF-κB Signaling Modulates miR-452-5p and miR-335-5p Expression to Functionally Decrease Epithelial Ovarian Cancer Progression in Tumor-Initiating Cells.

• DOI: 10.3390/ijms24097826
• 发表时间: 2023-04-25
• 期刊: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
• 影响因子: 5.6
• 作者: Kamdar, Rahul D.;Harrington, Brittney S.;Attar, Emma;Korrapati, Soumya;Shetty, Jyoti;Zhao, Yongmei;Tran, Bao;Wong, Nathan;House, Carrie D.;Annunziata, Christina M.
• 通讯作者: Annunziata, Christina M.