Nuclear Factor-kappaB in Ovarian Cancer

卵巢癌中的核因子-kappaB

• 批准号: 10926118
• 负责人: Christina Annunziata
• 金额: $ 97.96万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10926118
• 关键词: Adhesions; Affect; Aggressive behavior; Apoptosis; Biological; Biological Markers; Biology; Bypass; CASP8 gene; Cancer cell line; Cell Nucleus; Cell physiology; Cells; Chemoresistance; Chemotherapy-Oncologic Procedure; Classification; Clinic; Clinical; Clinical Trials; Collaborations; Complex; Cytoprotection; Databases; Dependence; Diagnosis; Disease; Drug Screening; Family; Gene Expression; Gene Expression Profile; Genes; Goals; Growth; Heterogeneity; Histone Deacetylase Inhibitor; I-kappa B Proteins; IKKepsilon; Immunofluorescence Immunologic; In Vitro; Invaded; Laboratories; Libraries; Lymphoid; Maintenance; Malignant Neoplasms; Malignant neoplasm of ovary; Mammary Neoplasms; Mediating; Molecular Structure; Multiple Myeloma; Mutation; NF-kappa B; NF-kappaB-inducing kinase; Neoplasm Metastasis; Nuclear; Outcome; Output; Pathogenesis; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Phase II Clinical Trials; Phenotype; Phosphorylation; Population; Predisposition; Process; Proliferating; Property; Proteins; Refractory; Relapse; Research; Resistance; Role; Signal Pathway; Signal Transduction; Solid Neoplasm; Specificity; TNFRSF5 gene; Testing; Therapeutic; Therapeutic Intervention; Time; Tissues; Woman; Work; cancer cell; cancer survival; chemotherapy; cytokine; design; development of lymphoid malignancy; effective therapy; extracellular; genetic manipulation; genetic signature; improved; inhibitor; insight; malignant breast neoplasm; mimetics; mouse model; multicatalytic endopeptidase complex; neoplastic cell; novel; ovarian neoplasm; overexpression; p65; pharmacologic; preclinical development; prognostic; prostate cancer cell; relapse prevention; response; small hairpin RNA; synergism; taxane; therapeutic target; transcription factor; translational approach; tumor; tumor initiation

The NF-kB pathway promotes survival of cancer cells. My research in ovarian cancer began with characterizing the activation state and biological relevance of NF-kB in this disease. The NF-kB family of transcription factors is ubiquitously expressed. NF-kB signaling has been implicated in ovarian cancer, but the significance and mechanism of NF-kB signaling in ovarian cancer is unknown. There is precedent to propose that NF-kB is a critical signaling mechanism in cancer. I initially hypothesized that the NF-kB pathway is over-activated in ovarian cancers with more aggressive behavior. The NF-kB pathway was implicated in ovarian cancer proliferation and cytokine secretion in vitro, and contributed to chemoresistance of ovarian cancer cell lines. I therefore sought to determine the expression patterns and prognostic associations of NF-kB pathway proteins in primary ovarian cancer tissues. I demonstrated that overexpression of the NF-kB subunit p50 at diagnosis conveyed poor outcome in these patients. The biological relevance of NF-kB in ovarian cancer was established in my laboratory. Having demonstrated the coordinate presence of NF-kB machinery in ovarian cancers, I sought to modulate its activity. Inhibitors of NF-kB (IkBs) are tagged for degradation through the proteasome upon specific inducible phosphorylation by IkB kinases (IKKs). Therefore, targeted inhibition of IKKs could isolate NF-kB as a mechanism for ovarian cancer pathogenesis. A subset of ovarian cancer cell lines was affected by inhibition of IKKb in properties of growth, adhesion, invasion and cytokine secretion. I developed a gene expression signature of IKKb signaling in ovarian cancer using both pharmacologic and genetic manipulation of IKKb. This signature gave insight into the results of NF-kB in ovarian cancer, based on known functions of the ovarian cancer-specific target genes, and allowed me to probe established ovarian cancer databases in order to estimate the relative impact of NF-kB signaling on the survival of women with ovarian cancer. Higher NF-kB activity conveyed a worse outcome, suggesting that modulation of IKKb might benefit patients whose tumors showed elevated target gene expression. A key discovery from this work was the tissue specificity of NF-kB signaling. The 9-gene signature experimentally defined in ovarian cancer was completely different from the 11 genes I previously identified in multiple myeloma. The overall goal of this project is to dissect the molecular structure of NF-kB signaling in ovarian cancer, with the intent to develop biomarkers of dependence on NF-kB, and novel points of therapeutic intervention. We completed two shRNA library screens, one in combination with an inhibitor of IKKbeta, and another in combination with shRNA against IKKepsilon. These studies identified novel interactions between the NF-kB pathway in ovarian cancer. In combination with IKKbeta, we found caspase 8 to be cooperative in protecting the cells from necroptosis. This work continues with designing a translational strategy for improving clinical outcome of women with caspase 8-deficient cancers. We plan to target the necroptosis pathway therapeutically, to bypass the defective apoptosis that mediates resistance to standard chemotherapy regimens. In a related avenue, we continue to study NF-kB signaling in ovarian cancer tumor-initiating cells. We observed by immunofluorescence that classical NF-kB appeared to be active in only a sub-population of the cultures at any given time, based on the presence of NF-kB p65 in the nucleus. Elevated classical NF-kB signaling has been observed in tumor-initiating cells (TICs) of prostate, breast, and ovarian tumors, but there are limited studies examining alternative NF-kB signaling. Both classical and alternative signaling cascades are required for maintenance and promotion of breast cancer TICs. The classical and alternative NF-kB pathways can regulate each other and integrate with other signaling pathways for fine-tuning functional outputs. Thus, the diverse and complex roles of NF-kB suggest this transcription factor family regulates cellular functions in a context dependent manner and may be a key factor in maintaining heterogeneity. We are currently testing the hypothesis that classical and alternative NF-kB pathways support distinct subpopulations of ovarian cancer tumor-initiating spheroids, that collaborate to populate secondary tumors following chemotherapy. Our recent results suggest that NF-kB supports the TIC phenotype and functional outputs of NF-kB are responsible for proliferation, chemoresistance, differentiation, multipotency. A better understanding of the biology underlying NF-kB signaling in TIC and non-TIC populations of tumor cells will guide the design of more effective therapies to overcome chemoresistance, prevent relapse, and improve survival of women with ovarian cancer. From a therapeutic standpoint, we previously completed a phase 2 clinical trial using the SMAC mimetic birinapant - that can target NF-kB signaling via inhibition of IAP proteins - in women with relapsed and refractory ovarian cancer. There were no clinical responses. Therefore, we completed a matrix drug screen testing 2000 compounds in combination with birinapant, in order to identify synergistic combinations to move to the clinic. We identified drug classes that met the criteria for synergy, and are in the process of testing these in mouse models. Ongoing work is testing the combination of SMAC mimetic with either HDAC inhibitor or Taxane chemotherapy in vitro and in mouse models. Three clinical trials are currently under development from this preclinical work.

NF-kB通路促进癌细胞存活。我对卵巢癌的研究始于表征NF-kB在这种疾病中的激活状态和生物学相关性。NF-kB转录因子家族是普遍表达的。NF-kB信号传导与卵巢癌有关，但NF-kB信号传导在卵巢癌中的意义和机制尚不清楚。有先例提出NF-kB是癌症的关键信号机制。我最初假设NF-kB通路在具有更具攻击性行为的卵巢癌中过度激活。NF-kB通路参与卵巢癌体外增殖和细胞因子分泌，并参与卵巢癌细胞系的化疗耐药。因此，我试图确定NF-kB途径蛋白在原发性卵巢癌组织中的表达模式和预后相关性。我证明了NF-kB亚基p50在诊断时的过表达在这些患者中传达了不良的结果。NF-kB在卵巢癌中的生物学相关性是在我的实验室建立的。在证明了NF-kB机制在卵巢癌中的协调存在后，我试图调节其活性。NF-kB抑制剂（IkBs）被标记为在IkB激酶（IKKs）特异性诱导磷酸化后通过蛋白酶体降解。因此，靶向抑制IKKs可以分离NF-kB作为卵巢癌发病机制之一。抑制IKKb可影响卵巢癌细胞系的生长、粘附、侵袭和细胞因子分泌。我利用IKKb的药理学和基因操作开发了卵巢癌中IKKb信号的基因表达特征。基于已知的卵巢癌特异性靶基因的功能，这一特征使我深入了解了NF-kB在卵巢癌中的结果，并使我能够探索已建立的卵巢癌数据库，以估计NF-kB信号传导对卵巢癌女性患者生存的相对影响。较高的NF-kB活性传达了较差的结果，这表明IKKb的调节可能有利于肿瘤中靶基因表达升高的患者。这项工作的一个关键发现是NF-kB信号的组织特异性。在卵巢癌中实验确定的9个基因特征与我之前在多发性骨髓瘤中确定的11个基因完全不同。该项目的总体目标是剖析卵巢癌中NF-kB信号的分子结构，旨在开发依赖NF-kB的生物标志物，以及新的治疗干预点。我们完成了两个shRNA文库筛选，一个与IKKbeta抑制剂联合，另一个与IKKepsilon的shRNA联合。这些研究发现了NF-kB通路在卵巢癌中的新的相互作用。结合IKKbeta，我们发现caspase 8可以协同保护细胞免于坏死。这项工作将继续设计一种转化策略，以改善患有caspase 8缺陷癌症的女性的临床结果。我们计划在治疗上靶向坏死下垂途径，以绕过介导对标准化疗方案耐药的缺陷细胞凋亡。在相关途径中，我们继续研究NF-kB信号在卵巢癌肿瘤启动细胞中的作用。我们通过免疫荧光观察到，基于细胞核中NF-kB p65的存在，在任何给定时间，经典NF-kB似乎只在培养的一个亚群中活跃。在前列腺、乳腺和卵巢肿瘤的肿瘤起始细胞（tic）中观察到经典NF-kB信号的升高，但对NF-kB信号的替代研究有限。经典和替代信号级联是维持和促进乳腺癌tic的必要条件。经典和替代NF-kB通路可以相互调节，并与其他信号通路整合以微调功能输出。因此，NF-kB的多样性和复杂作用表明，该转录因子家族以依赖于环境的方式调节细胞功能，可能是维持异质性的关键因素。我们目前正在测试一个假设，即经典的和可选择的NF-kB途径支持卵巢癌肿瘤启动球体的不同亚群，这些亚群在化疗后共同形成继发性肿瘤。我们最近的研究结果表明，NF-kB支持TIC表型，NF-kB的功能输出负责增殖、化学耐药、分化和多能性。更好地了解TIC和非TIC肿瘤细胞群中NF-kB信号传导的生物学基础，将指导设计更有效的治疗方法，以克服化疗耐药，防止复发，提高女性卵巢癌患者的生存率。从治疗的角度来看，我们之前在复发和难治性卵巢癌女性中完成了一项使用SMAC模拟双抗剂的2期临床试验，该双抗剂可以通过抑制IAP蛋白靶向NF-kB信号。没有临床反应。因此，我们完成了2000种化合物与birinapant联合使用的基质药物筛选测试，以确定协同组合，以便推向临床。我们确定了符合协同作用标准的药物类别，并正在小鼠模型中进行测试。正在进行的工作是在体外和小鼠模型中测试SMAC模拟物与HDAC抑制剂或紫杉烷化疗的组合。这项临床前工作目前正在进行三项临床试验。

项目成果

Topotecan synergizes with CHEK1 (CHK1) inhibitor to induce apoptosis in ovarian cancer cells.

• DOI: 10.1186/s12885-015-1231-z
• 发表时间: 2015-03-28
• 期刊: BMC cancer
• 影响因子: 3.8
• 作者: Kim MK;James J;Annunziata CM
• 通讯作者: Annunziata CM

Smac-mimetic enhances antitumor effect of standard chemotherapy in ovarian cancer models via Caspase 8-independent mechanism.

• DOI: 10.1038/s41420-021-00511-2
• 发表时间: 2021-06-04
• 期刊: Cell death discovery
• 影响因子: 7
• 作者: Hernandez LF;Dull AB;Korrapati S;Annunziata CM
• 通讯作者: Annunziata CM

BRD4 facilitates DNA damage response and represses CBX5/Heterochromatin protein 1 (HP1).

• DOI: 10.18632/oncotarget.17572
• 发表时间: 2017-08-01
• 期刊: Oncotarget
• 作者: Pongas G;Kim MK;Min DJ;House CD;Jordan E;Caplen N;Chakka S;Ohiri J;Kruhlak MJ;Annunziata CM
• 通讯作者: Annunziata CM

Molecular Pathways: The Balance between Cancer and the Immune System Challenges the Therapeutic Specificity of Targeting Nuclear Factor-κB Signaling for Cancer Treatment.

• DOI: 10.1158/1078-0432.ccr-15-1374
• 发表时间: 2016-09-01
• 期刊: Clinical cancer research : an official journal of the American Association for Cancer Research
• 作者: Zeligs KP;Neuman MK;Annunziata CM
• 通讯作者: Annunziata CM

Matrix Drug Screen Identifies Synergistic Drug Combinations to Augment SMAC Mimetic Activity in Ovarian Cancer.

• DOI: 10.3390/cancers12123784
• 发表时间: 2020-12-15
• 期刊: Cancers
• 影响因子: 5.2
• 作者: Noonan AM;Cousins A;Anderson D;Zeligs KP;Bunch K;Hernandez L;Shibuya Y;Goldlust IS;Guha R;Ferrer M;Thomas CJ;Annunziata CM
• 通讯作者: Annunziata CM