Elucidation of MLK3 Signaling in Hepatocellular Carcinoma

肝细胞癌中 MLK3 信号传导的阐明

• 批准号: 10324547
• 负责人: BASABI RANA
• 金额: --
• 依托单位: JESSE BROWN VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10324547
• 关键词: APC gene; APC mutation; Adult; Agonist; Alcoholism; Animals; BAY 54-9085; BCL2L1 gene; Cause of Death; Cell Cycle Arrest; Cell physiology; Ceramides; Childhood Liver Cancer; Combined Modality Therapy; Complex; Cyclin D1; Development; Diabetes Mellitus; Disease; Etiology; Event; FRAP1 gene; Family; Fatty Liver; Financial Hardship; Future; Genes; Genetic Diseases; Genetic Transcription; Goals; Hepatitis; Hepatitis B Virus; Hepatitis C virus; Human; In Vitro; Inflammation; Injury; Link; Liver Cirrhosis; Liver neoplasms; M cell; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of gastrointestinal tract; Mediating; Mediator of activation protein; Medical; Mitogen-Activated Protein Kinase Kinases; Modeling; Molecular; Mus; Mutation; Nuclear Translocation; Obesity; Oncogenic; PI3K/AKT; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphotransferases; Primary carcinoma of the liver cells; Proteins; Proto-Oncogene Proteins c-akt; Regulation; Reporting; Role; Signal Pathway; Signal Transduction; Signaling Protein; TCF Transcription Factor; Testing; Therapeutic; Tumor Suppressor Proteins; Veterans; Viral hepatitis; WNT Signaling Pathway; antagonist; base; beta catenin; c-myc Genes; cancer cell; chronic liver disease; design; effective therapy; glycogen synthase kinase 3 beta; in vivo; in vivo Model; inhibitor; insight; liver cancer model; military veteran; mixed lineage kinase 3; mutant; nanoparticle; new therapeutic target; next generation; non-alcoholic; nonalcoholic steatohepatitis; novel; overexpression; problem drinker; public health relevance; survivin; targeted treatment; therapy resistant; transcription factor

 DESCRIPTION (provided by applicant): Hepatocellular carcinoma (HCC) is one of the most common gastrointestinal (GI) malignancies and a major cause of death worldwide, with limited therapeutic options. Chronic liver diseases are believed to contribute significantly towards the development and progression of HCC due to their role in inflammation and injury. These chronic liver diseases including hepatitis, fatty live diseases (alcoholic and nonalcoholic) are prevalent among our veterans, thus creating a significant financial and medical burden. The only drug currently available to treat HCC is the multikinase inhibitor Sorafenib, which is effective only for a few months. Designing safe and efficient therapeutic approaches that can target additional signaling axes (aberrant in HCC) is critical for treating this deadly form of malignancy. Activation of various oncogenic pathways have been reported in HCC, which include PI3K/AKT/mTOR and Wnt/β-catenin signaling cascades, for which little or no effective therapies are available currently. Of particular importance is the Wnt/β-catenin pathway, since oncogenic mutations of β- catenin have been reported in a number of adult and childhood liver cancers. Attempts to develop drugs that can effectively target the Wnt/β-catenin axis has been extremely challenging mostly due to the fact that β- catenin is an intracellular protein without any enzymatic activity, and thus this axis is currently considered "undruggable". This is further complicated by the fact that β-catenin can localize in various cellular compartments and mediate multiple cellular functions some of which are paradoxical. Identification of specific upstream signaling pathways that can modulate β-catenin axis is thus critically needed to develop targeted therapies for HCC patients with aberrant activation of this axis. Our recent studies revealed a novel signaling pathway mediated by Mixed Lineage Kinase 3 (MLK3), a MAPK Kinase Kinase (MAP3K) that impacts β- catenin-mediated oncogenic signaling. Our studies indicate that MLK3 can phosphorylate and stabilize β- catenin in cancer cells, which involves a novel pathway independent of GSK3β. Despite stabilizing β-catenin, MLK3 inhibited β-catenin/TCF-dependent transcriptional activity and ultimately resulted in G2/M cell cycle arrest. Based on the preliminary results, we hypothesize that MLK3 activation can successfully antagonize HCC progression via inhibiting conventional Wnt/β-catenin pathway. The current proposal aims to validate our hypothesis utilizing both in vitro (cellular) and in vivo (animal) models while elucidating the fundamental mechanisms by which MLK3 modulates β-catenin pathway in HCC. The following specific aims are proposed to achieve our goals: (i) Determine the role of MLK3 in modulating Wnt/β-catenin axis in HCC cells, (ii) Elucidate the molecular mechanism of MLK3-induced modulation of Wnt/β-catenin axis in HCC cells and (iii) Determine the role of MLK3 in antagonizing Wnt/β-catenin axis in an in vivo model of HCC. Upon completion of these aims, we expect to establish the in vivo role of MLK3 in antagonizing Wnt/β-catenin axis in HCC, and identify the potential mechanism, which can be utilized for developing targeted therapeutic options in the future. The studies with the phospho-β-catenin mutants are expected to provide novel mechanistic insight revealing how this antagonism is achieved and identify new regulators of Wnt/β-catenin axis. Combined together, this information can be utilized to develop the next generation (more potent) MLK3 agonists to be utilized as monotherapy or in combination therapies to specifically target the β-catenin axis i HCC patients.

 描述（由申请人提供）： 肝细胞癌（HCC）是最常见的胃肠道（GI）恶性肿瘤之一，并且是全球死亡的主要原因，治疗方案有限。据信，慢性肝疾病由于其在炎症和损伤中的作用而对HCC的发展和进展产生重大贡献。这些慢性现场疾病包括肝炎，脂肪活疾病（酒精和非酒精性疾病）在我们的退伍军人中普遍存在，从而造成了重要的金融和医疗伯恩。目前唯一可用于治疗HCC的药物是多激酶抑制剂索拉非尼，这仅有效几个月。设计可以针对其他信号轴（HCC异常）的安全有效的治疗方法对于治疗这种致命的恶性肿瘤至关重要。 HCC中已经报道了各种致癌途径的激活，其中包括PI3K/AKT/MTOR和Wnt/β-catenin信号级联，目前几乎没有有效的疗法可用。 Wnt/β-catenin途径尤为重要，因为在许多成人和儿童肝癌中已经报道了β-catenin的致癌突变。试图开发可以有效靶向Wnt/β-catenin轴的药物的尝试极为挑战，主要是因为β-catenin是一种没有任何酶促活性的细胞内蛋白质，因此目前认为该轴“不可能”。 β-catenin可以定位在各种细胞室中并介导多个细胞功能的事实，这是更复杂的。因此，至关重要的是，需要确定可以调节β-catenin轴的特定上游信号通路，以开发针对该轴异常激活的HCC患者的靶向疗法。我们最近的研究揭示了由混合谱系激酶3（MLK3）介导的一种新型信号通路，这是一种MAPK激酶激酶（MAP3K），影响β-catenin介导的致癌信号传导。我们的研究表明，MLK3可以在癌细胞中磷酸化和稳定β-catenin，这涉及一种独立于GSK3β的新途径。尽管稳定β-catenin，MLK3仍抑制β-catenin/TCF依赖性转录活性，并最终导致G2/M细胞周期停滞。基于初步结果，我们假设MLK3激活可以通过抑制Wnt/β-catenin途径成功拮抗HCC的进展。当前的建议旨在通过体外（细胞）和体内（动物）模型验证我们的假设，同时阐明MLK3在HCC中调节β-catenin途径的基本机制。提出了以下具体目的来实现我们的目标：（i）确定MLK3在调节HCC细胞中Wnt/β-catenin轴上的作用，（ii）阐明MLK3诱导的Wnt/β-catenin轴在HCC细胞和（III）中的MLK3诱导的Wnt/β-catenin轴的调节的分子机制（III）确定MLK3在MLK3中的作用。 HCC。完成这些目标后，我们希望建立MLK3在HCC中对Wnt/β-catenin轴对抗的体内作用，并确定潜在机制，这些机制可以在将来开发出用于开发靶向治疗方案的潜在机制。对磷酸-β-catenin突变体的研究有望提供新颖的机械洞察力，揭示了如何实现这种拮抗作用并鉴定Wnt/β-catenin轴的新调节剂。合并在一起，可以利用这些信息来开发下一代（更多潜在的）MLK3激动剂，以作为单药治疗或联合疗法来专门针对β-catenin轴I HCC患者。

项目成果

Modulation of glycogen synthase kinase-3β following TRAIL combinatorial treatment in cancer cells.

• DOI: 10.18632/oncotarget.11834
• 发表时间: 2016-10-11
• 期刊: Oncotarget
• 作者: Santha S;Davaakhuu G;Basu A;Ke R;Das S;Rana A;Rana B
• 通讯作者: Rana B

Calcium channel blockers potentiate gemcitabine chemotherapy in pancreatic cancer.

• DOI: 10.1073/pnas.2200143119
• 发表时间: 2022-05-03
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1

Mixed Lineage Kinase 3 phosphorylates prolyl-isomerase PIN1 and potentiates GLI1 signaling in pancreatic cancer development.

• DOI: 10.1016/j.canlet.2021.04.015
• 发表时间: 2021-09-01
• 期刊: Cancer letters
• 影响因子: 9.7
• 作者: Viswakarma N;Sondarva G;Principe DR;Nair RS;Kumar S;Singh SK;Das S;Sinha SC;Grippo PJ;Grimaldo S;Giulianotti PC;Rana B;Rana A
• 通讯作者: Rana A

Involvement of AMP-activated protein kinase and Death Receptor 5 in TRAIL-Berberine-induced apoptosis of cancer cells.

• DOI: 10.1038/s41598-018-23780-x
• 发表时间: 2018-04-03
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Ke R;Vishnoi K;Viswakarma N;Santha S;Das S;Rana A;Rana B
• 通讯作者: Rana B

Ets1 mediates sorafenib resistance by regulating mitochondrial ROS pathway in hepatocellular carcinoma.

• DOI: 10.1038/s41419-022-05022-1
• 发表时间: 2022-07-04
• 期刊: CELL DEATH & DISEASE
• 影响因子: 9
• 作者: Vishnoi, Kanchan;Ke, Rong;Viswakarma, Navin;Srivastava, Piush;Kumar, Sandeep;Das, Subhasis;Singh, Sunil Kumar;Principe, Daniel R.;Rana, Ajay;Rana, Basabi
• 通讯作者: Rana, Basabi