Co-targeting MADD and Wnt/β-catenin signaling in Anaplastic Thyroid Cancer

甲状腺未分化癌中 MADD 和 Wnt/β-catenin 信号的共同靶向

• 批准号: 9885983
• 负责人: Bellur S Prabhakar
• 金额: --
• 依托单位: JESSE BROWN VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9885983
• 关键词: AKT Signaling Pathway; Ablation; Address; Aggressive behavior; Apoptosis; BAY 54-9085; BRAF gene; Basement membrane; CRISPR/Cas technology; CTNNB1 gene; Cessation of life; Clinical; Data; Death Domain; Distant; Drug Targeting; Drug resistance; Epithelial; Epithelial Cells; Epithelium; Exhibits; Gene Proteins; Growth; High Prevalence; Hyperactive behavior; In Vitro; Incidence; Knock-out; MAP Kinase Gene; Malignant Neoplasms; Malignant neoplasm of thyroid; Mediating; Mesenchymal; Metastatic Neoplasm to the Lung; Mitogen-Activated Protein Kinases; Mus; Mutation; Nature; Neoplasm Metastasis; Nodal; Normal Cell; Oncogenes; Operative Surgical Procedures; PTEN gene; Palliative Care; Pathway interactions; Patient Self-Report; Patients; Pharmaceutical Preparations; Phenotype; Phosphorylation; Population; Proteins; Proto-Oncogene Proteins c-akt; Publishing; Radiation exposure; Radioactive Iodine; Resistance; Role; Services; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; TNF gene; Testing; Treatment Efficacy; Tumor Suppressor Proteins; Tumor Tissue; Ubiquitination; Undifferentiated; United States; Veterans; WNT Signaling Pathway; advanced disease; agent orange; anaplastic thyroid cancer; base; beta catenin; cancer cell; effective therapy; high risk; in vivo; in vivo evaluation; inhibitor/antagonist; knock-down; mouse model; novel; novel therapeutics; outcome forecast; patient population; prevent; protein expression; protein function; standard care; stemness; targeted treatment; therapeutic development; therapeutic target; therapy resistant; treatment strategy; tumor; tumor growth

Project Description: Thyroid cancer incidence is rapidly increasing in the United States. Veterans are at even higher risk of developing thyroid cancer in their lifetime due to the increased potential of radiation exposure in the armed services. Furthermore, veterans who suffered from thyroid cancer self-reported a higher prevalence of Agent Orange exposure, increasing the relevance of this malignancy to the VA population. Despite the use of a range of combinations of treatments, ATC exhibits a dismal prognosis with a median survival of < six months. ATC tumors possess a greater mutation burden than all other forms of thyroid cancer which impart significant growth benefit and high metastatic potential Therefore, it is important to identify proteins that function at “nodal points” of different signaling pathways implicated in ATC, and thus could represent ATC “Achilles Heel.” The CTNNB1 (β-catenin) is an effector molecule of Wnt signaling which is critical for epithelial-mesenchymal transition (EMT) required for metastasis. PI3K/Akt/GSK3β signaling is hyperactive in ATC due to mutations. This can enhance β-catenin activity and also phosphorylate Map kinase Activating Death Domain-containing protein (MADD) and contribute to its pro-survival function. pMADD renders ATC cells resistant to apoptosis. Thus in ATC, pMADD and β-catenin can serve as two potential “nodal points.” Most importantly, our recent novel findings have shown that MADD knockdown can significantly inhibit TNFα mediated activation of β-catenin signaling by preventing pERK activation and consequent pGSK3β activation. Lack of GSK3β phosphorylation by ERK, facilitated ubiquitination of β-catenin leading to its degradation and resultant blockage of EMT activation. Furthermore, intra-tumoral administration of MADD siRNA significantly reduced orthotpic ATC tumor growth and lung metastasis in treated mice. Therefore, MADD is a potential therapeutic target in ATC either alone or in combination with Wnt/β-catenin inhibitors. Based on a very strong scientific premise, we hypothesize that MADD down modulation can be effective in inhibiting growth and overcoming resistance to drugs targeting hyperactive MAPK, PI3K/Akt and Wnt/β-catenin signaling, which are hallmarks of ATC. To address this, in aim-1, we will functionally characterize the impact of CRISPR/CAS9 mediated MADD gene knock-out in ATC cells on Wnt signaling in vitro and ex vivo; in aim-2, we will determine the impact of down-modulating MADD expression, and MAPK and PI3K/Akt signaling pathways, on Wnt signaling; and in aim-3, we will test the in vivo therapeutic efficacy of combination treatment with MADD knockdown in orthotopic and spontaneous mouse models of ATC. ATC disproportionately accounts for the majority of thyroid cancer-related deaths. Cancer cell-specific expression of MADD, its role in activating several key signaling pathways, and its ability to act as a pro-survival factor and promote metastasis in ATC makes it an ideal target for therapeutic development. Importantly, MADD deletion had no apparent effect on normal cells. Thus the proposed novel studies are highly relevant to veterans.

项目描述：美国的甲状腺癌发病率正在迅速增加。退伍军人处于平局 由于辐射暴露的可能性增加，一生中患甲状腺癌的风险更高 武装部队。此外，患有甲状腺癌的退伍军人自我报告的患病率更高 橙剂暴露，增加了这种恶性肿瘤与退伍军人管理局人群的相关性。尽管使用 通过一系列治疗组合，ATC 的预后较差，中位生存期< 6 个月。 ATC 肿瘤比所有其他形式的甲状腺癌具有更大的突变负担，这会导致显着的突变。 生长效益和高转移潜力因此，识别在“节点”发挥作用的蛋白质非常重要 ATC 涉及的不同信号通路的“点”，因此可以代表 ATC 的“致命弱点”。这 CTNNB1（β-连环蛋白）是 Wnt 信号传导的效应分子，对上皮间质细胞至关重要 转移所需的转变（EMT）。由于突变，ATC 中 PI3K/Akt/GSK3β 信号传导过度活跃。这 可以增强 β-catenin 活性，还可以磷酸化含有 Map 激酶激活死亡结构域的蛋白质 （MADD）并有助于其促进生存的功能。 pMADD 使 ATC 细胞抵抗细胞凋亡。因此在 ATC、pMADD 和 β-catenin 可以作为两个潜在的“节点”。 最重要的是，我们最近的新发现表明 MADD 敲低可以显着抑制 TNFα 通过阻止 pERK 激活和随后的 pGSK3β 介导 β-连环蛋白信号传导的激活 激活。 ERK 缺乏 GSK3β 磷酸化，促进 β-catenin 泛素化，导致其 降解并由此导致 EMT 激活受阻。此外，MADD siRNA 的肿瘤内给药 显着减少治疗小鼠的正位 ATC 肿瘤生长和肺转移。因此，MADD 是一个 ATC 的潜在治疗靶点，无论是单独使用还是与 Wnt/β-连环蛋白抑制剂联合使用。 基于非常有力的科学前提，我们假设 MADD 下调制可能是有效的 抑制生长并克服针对高活性 MAPK、PI3K/Akt 和 Wnt/β-catenin 的药物的耐药性 信令，这是 ATC 的标志。为了解决这个问题，在目标 1 中，我们将从功能上描述以下因素的影响： CRISPR/CAS9 介导体外和离体 ATC 细胞中 Wnt 信号传导 MADD 基因敲除；在目标2中，我们 将确定下调 MADD 表达以及 MAPK 和 PI3K/Akt 信号通路的影响， 关于 Wnt 信号传导；在aim-3中，我们将测试与MADD联合治疗的体内疗效 ATC 的原位和自发小鼠模型中的敲低。 ATC 不成比例地占甲状腺癌相关死亡的大部分。癌细胞特异性 MADD 的表达、其在激活几个关键信号通路中的作用及其作为促生存因子的能力 因素并促进 ATC 转移，使其成为治疗开发的理想靶点。重要的是，MADD 缺失对正常细胞没有明显影响。因此，拟议的新颖研究与退伍军人高度相关。