Combinatorial Approach for Reducing Invasive Potential of Melanoma Cells

降低黑色素瘤细胞侵袭潜力的组合方法

• 批准号: 8510083
• 负责人: FARRUKH AFAQ
• 金额: $ 19.12万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8510083
• 关键词: Adverse effects; BAY 54-9085; BRAF gene; Cell Line; Cells; Cessation of life; Clinical Research; Collagen; Data; Development; Dietary Flavonoid; Dose; Drug resistance; Epithelial; Epithelial Cells; Fibroblasts; Genetic; Green Fluorescent Proteins; Health; Human; In Vitro; Lead; Life; Liver; Lung; MEKs; Malignant Neoplasms; Melanoma Cell; Mesenchymal; Mesentery; Metastatic Melanoma; Mitogen-Activated Protein Kinases; Molecular; Mutation; Neoplasm Metastasis; Nude Mice; Oncogenic; Organ; PI3K/AKT; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phenotype; Phosphorylation; Phytochemical; Prevention; Prevention strategy; Process; Property; Proto-Oncogene Proteins c-akt; Publishing; Regimen; Resistance; Signal Pathway; Signal Transduction; Skin; Skin Cancer; Survival Rate; Tail; Testing; Therapeutic; Tissue Engineering; Tissues; Toxic effect; Treatment Efficacy; Veins; bone; cancer cell; cancer recurrence; clinically relevant; combinatorial; design; fisetin; fruits and vegetables; improved; in vivo; inhibitor/antagonist; intravenous injection; keratinocyte; lymph nodes; melanoma; novel; outcome forecast; programs; protein expression; public health relevance; research study; response; subcutaneous

DESCRIPTION (provided by applicant): Melanoma is one of the fastest-rising malignancies in the past few decades. It is a serious form of cancer responsible for approximately 80% of skin cancer-related deaths. Melanoma is resistant to current therapeutic regimens because for its propensity to metastasize and has a very poor prognosis with a median survival rate of six months. Studies have demonstrated that the majority of human melanomas have constitutively active mitogen activated protein kinases (MAPK) through oncogenic mutation in either BRAF or NRAS. In malignant melanoma the BRAF-MEK-ERK and PI3K-AKT-NFkB signaling pathways are constitutively activated and regulated by RAS signals. Activation of these pathways leads to an induction of epithelial-mesenchymal transition (EMT) resulting in cell invasion. EMT is a molecular program whereby epithelial cells undergo reprogramming from a polarized differentiated phenotype to a mesenchymal phenotype and is considered to be a key process in regulating the initial step of metastatic progression. Targeting BRAF with sorafenib resulted in inhibition of MAPK signaling pathways both in vitro and in vivo. However, as a monotherapy it is not effective in patients with metastatic melanoma as revealed by a phase II clinical study. These data and other published studies support the hypothesis that it is not sufficient to inhibit only a single constitutively activated signaling pathway for the treatment of melanoma. Recently, we found that fisetin treatment reduced melanoma cell invasion by inhibiting the protein expression of PI3K (p110a and p85) and phosphorylation of AKT at Ser473 and Thr308. It also reduced invasive potential of metastatic melanoma A375 cells in tissue-engineered three dimensional skin equivalents. Furthermore, fisetin treatment was found to reduce the formation of A375 cell colonies. The present proposal capitalizes on our recent novel unpublished findings and is designed to investigate the effects of fisetin alone and in combination with sorafenib on invasion and metastasis of melanoma cells by targeting BRAF and PI3K/AKT pathways. The central hypothesis to be tested in this proposal is that the "combination of fisetin with sorafenib will be more effective in reducing melanoma cell invasion and metastasis than with fisetin or sorafenib alone". To test our hypothesis, two inter-related specific aims are proposed: (I) to investigate the effect of fisetin alone and in combination with sorafenib on invasion and EMT in melanoma cell lines and in tissue-engineered three dimensional skin equivalents by targeting BRAF and PI3K/AKT signaling pathways, and (II) to determine the effect of fisetin alone and in combination with sorafenib on the formation of systemic metastases in athymic Crl:NU-Foxn1nu nude mice by an intravenous injection of GFP-tagged melanoma cells. HEALTH RELEVANCE: Through this proposal we envision the identification of a natural nontoxic dietary flavonoid fisetin, which can be exploited in combination with sorafenib for the prevention of cell invasion and metastasis from which a large number of patients would stand to benefit.

描述(申请人提供)：黑色素瘤是过去几十年中增长最快的恶性肿瘤之一。它是一种严重的癌症，大约80%的皮肤癌相关死亡是由它造成的。黑色素瘤对目前的治疗方案有抵抗力，因为它有转移的倾向，预后非常差，中位存活率为6个月。研究表明，大多数人类黑色素瘤通过BRAF或NRAS中的致癌基因突变而具有结构性活性的丝裂原激活蛋白激酶(MAPK)。在恶性黑色素瘤中，BRAF-MEK-ERK和PI3K-AKT-NFkB信号通路受RAS信号的结构性激活和调节。这些通路的激活导致上皮-间充质转化(EMT)的诱导，从而导致细胞侵袭。EMT是一种分子程序，使上皮细胞经历从极化分化表型到间充质表型的重新编程，被认为是调节转移进展的初始步骤的关键过程。索拉非尼靶向BRAF在体外和体内均可抑制MAPK信号转导通路。然而，作为单一疗法，它对II期临床研究揭示的转移性黑色素瘤患者并不有效。这些数据和其他已发表的研究支持这样的假设，即仅抑制单一的结构性激活的信号通路用于治疗黑色素瘤是不够的。最近，我们发现，非瑟素治疗通过抑制PI3K(p110a和p85)的蛋白表达和AKT在Ser473和Thr308的磷酸化来减少黑色素瘤细胞的侵袭性。它还降低了转移性黑色素瘤A375细胞在组织工程三维皮肤等效物中的侵袭潜力。此外，还发现非瑟素处理可减少A375细胞集落的形成。目前的建议利用我们最近未发表的新发现，旨在通过靶向BRAF和PI3K/AKT通路来研究非瑟素单独和联合索拉非尼对黑色素瘤细胞侵袭和转移的影响。在这项提议中要检验的中心假设是：“非瑟丁与索拉非尼的联合 在减少黑色素瘤细胞侵袭和转移方面将比单独使用非赛汀或索拉非尼更有效。为了验证我们的假设，我们提出了两个相互关联的特定目标：(I)通过靶向BRAF和PI3K/AKT信号通路，研究非瑟素单独及联合索拉非尼对黑色素瘤细胞系和组织工程三维皮肤等效物侵袭和EMT的影响；(Ii)通过静脉注射GFP标记的黑色素瘤细胞，确定非瑟素单独及联合索拉非尼对裸鼠无瘤CRL：NU-Foxn1nu裸鼠全身转移的影响。健康相关性：通过这项建议，我们设想鉴定一种天然的无毒膳食类黄酮非瑟素，它可以与索拉非尼联合开发用于预防细胞侵袭和转移，将使大量患者受益。