Selective KGFR Antagonists for the Prevention of Cancer Metastasis

选择性 KGFR 拮抗剂预防癌症转移

• 批准号: 7365010
• 负责人: JOSEPH Thomas PENTO
• 金额: $ 21.98万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7365010
• 关键词: Actins; Address; Adult; Affinity; Animals; Area; Binding; Biological; Biological Assay; Biology; Breast; Breast Cancer Cell; Breast Carcinoma; Cell Proliferation; Cells; Class; Computer Assisted; Development; Dose; Ductal Breast Hyperplasia; Epidermal Growth Factor Receptor; Fatty acid glycerol esters; Female; Frozen Sections; Gefitinib; Growth; Growth and Development function; Harvest; Human; Implant; In Vitro; Libraries; Liver; Lung; Lung Neoplasms; MCF7 cell; Malignant Neoplasms; Mammary gland; Measures; Mediating; Membrane; Methods; Microscopy; Modeling; Mus; National Cancer Institute; Neoplasm Metastasis; Nude Mice; Prevention; Range; Receptor Inhibition; Receptor Signaling; Relative (related person); Reporting; Research; Research Personnel; Rodent; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Specificity; Testing; Therapeutic Agents; Time; Tyrosine Kinase Inhibitor; Video Microscopy; Western Blotting; Xenograft Model; Xenograft procedure; base; cancer cell; cancer type; cell motility; day; density; design; dosage; in vivo; inhibitor/antagonist; keratinocyte growth factor; keratinocyte growth factor receptor; malignant breast neoplasm; migration; novel; prevent; protein expression; receptor; receptor binding; research study; therapeutic target; tumor growth; tumor progression; tumor xenograft

DESCRIPTION (provided by applicant): Background: The mammary glands of adult female animals are remarkably sensitive to KGF. KGF acts at the KGFR to produce a rapid and profound stimulation of breast cancer cell proliferation and motility. Further, KGF-induced motility in breast cancer cells is mediated via the Erk1/2 signaling pathway. Thus, enhancement of KGF/KGFR signal transduction may be an early step in the metastatic progression of breast cancer. Receptor modeling of KGFR was used to identify a library of selective KGFR TKI molecules with high receptor affinity. Forty of the KGFR TKI have been synthesized and are available to study the involvement of KGFR signaling in breast cancer metastasis. Hypothesis: KGFR is an important therapeutic target in breast cancer; and thus, selective inhibition of KGFR-mediated signaling will reduce or eliminate breast cancer cell proliferation, motility and metastasis. Specific Aims: The First Specific Aim will measure the relative activity and specificity of the 40 KGFR TKI on KGF-mediated KGFR signal transduction in human breast cancer cells. The Second Specific Aim will examine the ability of these 40 inhibitors to reduce KGF-induced breast cancer cell proliferation and motility in vitro. The Third Specific Aim will determine the inhibitory effect of the most selective and potent KGFR TKI (as determined in vitro in Specific Aims #1 and 2) on KGF-mediated growth and metastasis of human breast cancer cells in a mouse xenograft model. Methods: In Specific Aim #1, MCF-7 breast cancer cells will receive either a motility stimulating dose of KGF (50 ng/ml) or KGFR TKI at doses ranging from 10 to 200 ¿M or KGF (50 ng/ml) + KGFR TKI at each inhibitor dose or Iressa (epidermal growth factor receptor selective TK inhibitor; used as a negative control) over the same dosage range or KGF + Iressa (at the same doses) or vehicle control. At various times, from 1 to 48 hrs following KGF administration, the cells will be harvested to determine the protein expression of KGFR, phospho-KGFR, Erk1/2, phospho-Erk1/2 and ¿-actin by Western blotting. In Specific Aim #2 proliferation and migration of MCF-7 cells will be evaluated with both time-lapse videomicroscopy and a culture wounding assay over a period of 1 to 3 days using the same experimental groups and doses described in the first specific aim. In Specific Aim #3 the influence of KGFR inhibition on the growth and metastasis of tumor xenografts will be determined. The xenograft studies will be conducted by implanting MCF-7 cells into mammary fat pads of nude mice. At the end of the experiment, tumor, lung and liver will be removed and frozen sections examined by fluorescent microscopy to quantify tumor growth and development of lung and liver micro-metastasis. Significance: The establishment of KGFR as an important therapeutic target, together with the identification of specific and potent KGFR TKI, should result in the rapid development of a new class of highly selective therapeutic agents to prevent the metastatic progression of cancer. The establishment of KGFR as an important therapeutic target, together with the identification of specific and potent KGFR TKI, should result in the rapid development of a new class of highly selective therapeutic agents designed to prevent the metastatic progression of cancer. This project addresses several focus areas of the National Cancer Institute; namely, the biology of cancer metastasis and the development of highly selective and molecularly targeted therapeutic agents.

描述（由申请人提供）： 背景：成年雌性动物的乳腺对 KGF 非常敏感。 KGF 作用于 KGFR，对乳腺癌细胞增殖和运动产生快速而深刻的刺激。此外，KGF 诱导的乳腺癌细胞运动是通过 Erk1/2 信号通路介导的。因此，KGF/KGFR信号转导的增强可能是乳腺癌转移进展的早期步骤。 KGFR 受体模型用于鉴定具有高受体亲和力的选择性 KGFR TKI 分子库。四十种 KGFR TKI 已被合成，可用于研究 KGFR 信号传导在乳腺癌转移中的作用。假设：KGFR是乳腺癌的重要治疗靶点；因此，选择性抑制 KGFR 介导的信号传导将减少或消除乳腺癌细胞的增殖、运动和转移。具体目标：第一个具体目标将测量 40 KGFR TKI 对人乳腺癌细胞中 KGF 介导的 KGFR 信号转导的相对活性和特异性。第二个具体目标将检查这 40 种抑制剂在体外减少 KGF 诱导的乳腺癌细胞增殖和运动的能力。第三个具体目标将确定最具选择性和最有效的 KGFR TKI（如具体目标 #1 和 2 中的体外测定）对小鼠异种移植模型中 KGF 介导的人乳腺癌细胞生长和转移的抑制作用。方法：在具体目标 #1 中，MCF-7 乳腺癌细胞将接受运动刺激剂量的 KGF (50 ng/ml) 或剂量范围为 10 至 200 µM 的 KGFR TKI，或每个抑制剂剂量的 KGF (50 ng/ml) + KGFR TKI，或相同剂量的易瑞沙（表皮生长因子受体选择性 TK 抑制剂；用作阴性对照） 范围或 KGF + 易瑞莎（相同剂量）或载体对照。在施用KGF后1至48小时的不同时间，收获细胞以通过Western印迹测定KGFR、磷酸-KGFR、Erk1/2、磷酸-Erk1/2和β-肌动蛋白的蛋白质表达。在具体目标 #2 中，将使用第一个具体目标中描述的相同实验组和剂量，在 1 至 3 天内通过延时视频显微镜和培养物损伤测定来评估 MCF-7 细胞的增殖和迁移。在具体目标#3中，将确定KGFR抑制对肿瘤异种移植物生长和转移的影响。异种移植研究将通过将 MCF-7 细胞植入裸鼠的乳腺脂肪垫进行。实验结束时，将切除肿瘤、肺和肝脏，并通​​过荧光显微镜检查冷冻切片，以量化肿瘤生长以及肺和肝脏微转移的发展。意义：KGFR 作为重要治疗靶点的确立，加上特异有效的 KGFR TKI 的鉴定，应该会导致一类新型高选择性治疗药物的快速开发，以预防癌症的转移进展。 KGFR 作为重要治疗靶点的确立，加上特异有效的 KGFR TKI 的鉴定，应该会导致一类新型高选择性治疗药物的快速开发，旨在预防癌症的转移进展。该项目涉及国家癌症研究所的几个重点领域；即癌症转移的生物学以及高选择性和分子靶向治疗剂的开发。

项目成果

Influence of novel KGFR tyrosine kinase inhibitors on KGF-mediated proliferation of breast cancer.

• 发表时间: 2010-12
• 期刊: Anticancer research
• 影响因子: 2
• 作者: Meghna Mehta;J. Kesinger;X. Zang;M. Lerner;D. Brackett;R. Brueggemeier;Pui-Kui Li;J. Pento

Oncolytic potential of a novel KGFR tyrosine kinase inhibitor using a KGFR-selective breast cancer xenograft model.

使用 KGFR 选择性乳腺癌异种移植模型研究新型 KGFR 酪氨酸激酶抑制剂的溶瘤潜力。

• 发表时间: 2015
• 期刊: Anticancer research
• 影响因子: 2
• 作者: Kesinger,JasonW;Mehta,Meghna;Lerner,MeganR;Brackett,DanielJ;Brueggemeier,RobertW;Li,Pui-Kui;Pento,JThomas
• 通讯作者: Pento,JThomas