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: 10618953
• 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 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10618953
• 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; Combined Modality Therapy; Data; Death Domain; Distant; Drug Targeting; Drug resistance; Epithelial Cells; Epithelium; Exhibits; Gene Proteins; Growth; High Prevalence; Hyperactivity; In Vitro; Incidence; 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; PIK3CG gene; PTEN gene; Palliative Care; Pathway interactions; Patient Self-Report; Patients; Pharmaceutical Preparations; Phenotype; Phosphorylation; Population; Prognosis; 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; beta catenin; cancer cell; effective therapy; glycogen synthase kinase 3 beta; high risk; in vivo; in vivo evaluation; inhibitor; kinase inhibitor; knock-down; knockout gene; migration; mouse model; novel; novel therapeutics; 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：美国甲状腺癌发病率正在迅速增加。退伍军人在 由于辐射暴露的潜力增加，甲状腺癌在其一生中较高的风险更高 武装服务。此外，患有甲状腺癌的退伍军人自我报告较高的患病率 特工橙色暴露，增加了这种恶性肿瘤与VA人群的相关性。尽管使用了 ATC的一系列治疗组合表现出令人沮丧的预后，中位生存期为六个月。 与所有其他形式的甲状腺癌相比，ATC肿瘤具有更大的突变燃烧 因此，生长益处和高转移性潜力，重要的是鉴定在淋巴结中起作用的蛋白质 ATC中实现的不同信号通路的点”，因此可以代表ATC“阿喀琉斯高跟鞋”。这 CTNNB1（β-catenin）是Wnt信号的效应分子，对上皮 - 间质至关重要 转移所需的过渡（EMT）。由于突变，PI3K/AKT/GSK3β信号传导在ATC中过度活跃。这 可以增强β-catenin活性，还可以磷酸化的MAP激酶激活含死亡结构域的蛋白 （MADD）并促进其亲生功能。 PMADD使ATC细胞抗凋亡。那在 ATC，PMADD和β-catenin可以用作两个潜在的“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介导的β-catenin信号传导的激活通过预防PERK激活和随之而来的PGSK3β激活 激活。 ERK缺乏GSK3β磷酸化，制备了β-catenin的泛素化导致其 EMT激活的降解和结果阻塞。此外，MADD siRNA的肿瘤内管理 在治疗的小鼠中，明显降低了正直ATC肿瘤的生长和肺转移。因此，Madd是一个 单独或与Wnt/β-catenin抑制剂组合的ATC中的潜在治疗靶标。 基于一个非常强大的科学前提，我们假设MADD降低调制可能是有效的 在抑制生长并克服对靶向多动MAPK的药物，PI3K/AKT和WNT/β-catenin的抗性 信号，是ATC的标志。为了解决这个问题，在AIM-1中，我们将在功能上表征 CRISPR/CAS9在体外和Ex Vivo中介导了ATC细胞中ATC细胞中的MADD基因敲除；在AIM-2中，我们 将确定下调MADD表达的影响以及MAPK和PI3K/AKT信号通路， 在Wnt信号上；在AIM-3中，我们将测试与MADD组合治疗的体内治疗效率 ATC的原位和赞助小鼠模型中的敲低。 ATC不成比例地解释了大多数甲状腺癌相关的死亡。癌细胞特异性 MADD的表达，其在激活多种关键信号通路中的作用及其充当生存的能力 因素并促进ATC中的转移使其成为治疗性发育的理想目标。重要的是，Madd 缺失对正常细胞没有明显的影响。拟议的新研究与退伍军人高度相关。