A role for AKT2 in BRAFV600E melanoma initiation, progression, and response to therapy

AKT2 在 BRAFV600E 黑色素瘤发生、进展和治疗反应中的作用

基本信息

批准号：
9320005
负责人：
Siobhan K. Mcree
金额：
$ 3.58万
依托单位：
TUFTS UNIVERSITY BOSTON
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-16 至 2018-05-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9320005
关键词：
AKT Signaling Pathway AKT inhibition AKT1 gene AKT2 gene AKT3 gene Ablation Aggressive behavior Allografting Antibodies Automobile Driving BRAF gene Cell Line Cells Classification Clinical Trials Data Diagnostic Disease Disease Progression Dose-Limiting Drug Targeting Drug resistance Early Diagnosis Extravasation Family Future Gene Amplification Genes Genetic Genomics Goals Human In Vitro Injection of therapeutic agent Investigation MAP Kinase Signaling Pathways Malignant Neoplasms Mass Spectrum Analysis Mediating Mediator of activation protein Melanoma Cell Metabolism Metastatic Melanoma Mitogen-Activated Protein Kinases Morphology Mus Mutation Neoplasm Metastasis Null Lymphocytes Oncogenic Outcome PI3K/AKT Pathway interactions Pharmaceutical Preparations Phenotype Phosphoproteins Phosphotransferases Play Process Protein Isoforms Protein-Serine-Threonine Kinases Proto-Oncogene Proteins c-akt Recurrence Resistance Role Skin Cancer Specificity Testing Therapeutic Therapeutic Intervention Toxic effect Treatment Protocols Tumor Burden Ursidae Family Work cancer cell cell growth cell motility epithelial to mesenchymal transition experimental study improved in vivo inhibitor/antagonist kinase inhibitor knock-down melanocyte melanoma migration mouse model mutant novel novel therapeutic intervention novel therapeutics outcome forecast resistance mechanism response targeted treatment therapeutic candidate treatment response tumor

AKT Signaling Pathway AKT inhibition AKT1 gene AKT2 gene AKT3 gene Ablation Aggressive behavior Allografting Antibodies Automobile Driving BRAF gene Cell Line Cells Classification Clinical Trials Data Diagnostic Disease Disease Progression Dose-Limiting Drug Targeting Drug resistance Early Diagnosis Extravasation Family Future Gene Amplification Genes Genetic Genomics Goals Human In Vitro Injection of therapeutic agent Investigation MAP Kinase Signaling Pathways Malignant Neoplasms Mass Spectrum Analysis Mediating Mediator of activation protein Melanoma Cell Metabolism Metastatic Melanoma Mitogen-Activated Protein Kinases Morphology Mus Mutation Neoplasm Metastasis Null Lymphocytes Oncogenic Outcome PI3K/AKT Pathway interactions Pharmaceutical Preparations Phenotype Phosphoproteins Phosphotransferases Play Process Protein Isoforms Protein-Serine-Threonine Kinases Proto-Oncogene Proteins c-akt Recurrence Resistance Role Skin Cancer Specificity Testing Therapeutic Therapeutic Intervention Toxic effect Treatment Protocols Tumor Burden Ursidae Family Work cancer cell cell growth cell motility epithelial to mesenchymal transition experimental study improved in vivo inhibitor/antagonist kinase inhibitor knock-down melanocyte melanoma migration mouse model mutant novel novel therapeutic intervention novel therapeutics outcome forecast resistance mechanism response targeted treatment therapeutic candidate treatment response tumor

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项目摘要

Project Summary Despite recent advances, melanoma remains the deadliest form of skin cancer, and new therapeutic strategies are urgently needed. The MAP kinase (MAPK) and PI3K/AKT pathways are central to both disease progression and chemoresistance in melanoma, with dual targeting approaches yielding promising results that would benefit from increased target specificity and selectivity. The AKT family of serine/threonine kinases comprises three highly homologous and functionally distinct isoforms (AKT1, AKT2, and AKT3) that play unique roles in melanoma but have not yet been leveraged for isoform-specific targeting. AKT2 is an attractive candidate for therapeutic intervention, given that AKT2 mutations and AKT2 gene amplification are known mechanisms of adaptive resistance to current targeted therapies such as the BRAF inhibitor (BRAFi) Vemurafenib. We hypothesize that AKT2 is involved in melanoma invasion and metastasis, and acts as a mediator of the BRAFi response. We further suggest that testing AKT2 isoform-specific targeting in combination with BRAF inhibitors is an anti-melanoma strategy that may avoid serious dose limiting toxicities commonly associated with pan-AKT inhibition. This work will establish the contribution of AKT2 to BRAF-mutant melanoma progression and metastasis, using both human melanoma cell lines and mouse models. In Aim1, we will investigate the effect of AKT2 knockdown on human melanoma cell migration and invasion, and whether this differs in the presence of the BRAFi Vemurafenib. Further, we will identify AKT2-specific substrates that influence the BRAFi response by immunoprecipitating phosphoproteins reacting with AKT phospho-substrate-specific antibodies in cells treated with Vemurafenib with or without AKT2-specific knockdown. Immunoprecipitates will be analyzed by mass spectrometry, prioritized and validated using AKT2 wild-type and knockdown cells to determine if manipulation of any substrate alters cell sensitivity to BRAF inhibition. In Aim 2, we will investigate the contribution of genetic AKT2 loss in a BRAF-mutant melanoma prone mouse model, and perform metastasis seeding experiments by intra-cardiac injection of AKT2 null cell lines, as well as drug studies in allograft tumors. We will investigate the effect of co-targeting BRAF and AKT2 with specific inhibitors, as a proof-of- principle for future studies. With AKT inhibitors in clinical trials for melanoma, our findings could yield novel therapeutic strategies and increase the efficiency of existing therapies to improve treatment options and outcome for this devastating disease.

项目摘要尽管最近进展，黑色素瘤仍然是皮肤癌和新治疗的最致命形式迫切需要策略。地图激酶（MAPK）和PI3K/AKT途径对于这两种疾病都是核心黑色素瘤的进展和化学抗性，双重靶向方法产生了令人鼓舞的结果目标特异性和选择性的提高将受益。 AKT丝氨酸/苏氨酸激酶家族包括三个高度同源且功能上不同的同工型（AKT1，AKT2和AKT3）在黑色素瘤中的独特作用，但尚未利用同工型特异性靶向。 AKT2很有吸引力鉴于已知AKT2突变和Akt2基因扩增的治疗干预术的候选者对当前靶向疗法（例如BRAF抑制剂）的适应性抗性机制（BRAFI） vemurafenib。我们假设AKT2参与了黑色素瘤侵袭和转移，并充当 Brafi响应的中介。我们进一步建议，在与BRAF抑制剂结合是一种抗黑色素瘤策略，可能避免严重限制毒性通常与Pan-Akt抑制相关。这项工作将确定Akt2对BRAF突变的黑色素瘤进展和转移，使用人黑色素瘤细胞系和小鼠模型。在AIM1中，我们将研究 AKT2敲除人类黑色素瘤细胞迁移和侵袭，以及在存在的情况下这是否有所不同 Brafi Vemurafenib。此外，我们将确定影响BRAFI响应的Akt2特异性底物通过免疫沉淀与细胞中Akt磷酸基底特异性抗体反应的磷酸蛋白在有或没有AKT2特异性敲低的情况下用vemurafenib处理。免疫沉淀将通过质谱法，使用AKT2野生型和敲低细胞进行优先级和验证，以确定是否是否操纵任何底物都会改变细胞对BRAF抑制的敏感性。在AIM 2中，我们将调查 BRAF突变的黑色素瘤易于小鼠模型中遗传AKT2损失的贡献，并进行转移播种实验通过对Akt2无效细胞系的心内注射以及同种异体移植中的药物研究肿瘤。我们将研究与特定抑制剂共同靶向BRAF和AKT2的效果，作为证明未来研究的原则。在黑色素瘤临床试验中，AKT抑制剂可以产生新颖治疗策略并提高现有疗法的效率，以改善治疗选择和这种毁灭性疾病的结果。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

AKT2 Loss Impairs BRAF-Mutant Melanoma Metastasis.

DOI：
10.3390/cancers15204958
发表时间：
2023-10-12
期刊：
CANCERS
影响因子：
5.2
作者：
Mcree, Siobhan K.;Bayer, Abraham L.;Pietruska, Jodie;Tsichlis, Philip N.;Hinds, Philip W.
通讯作者：
Hinds, Philip W.