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（适用提供）：背景：成年雌性动物的乳腺对kgf非常敏感。 KGF在KGFR上起作用，对乳腺癌细胞增殖和运动性产生快速而深刻的刺激。此外，KGF诱导的乳腺癌细胞中的运动性是通过ERK1/2信号通路介导的。这是KGF/KGFR信号转导的增强可能是乳腺癌转移性进展的早期一步。 KGFR的受体建模用于识别具有高受体亲和力的选择性KGFR TKI分子的库。四十KGFR TKI已合成，可用于研究KGFR信号传导参与乳腺癌转移。假设：KGFR是乳腺癌中重要的治疗靶标。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。具体目的：第一个具体目标将测量40 kgfr tki在人类乳腺癌细胞中KGF介导的KGFR信号转移中的相对活性和特异性。第二个特定目的将检查这40种抑制剂减少KGF诱导的乳腺癌细胞增殖和体外运动的能力。第三个特定目的将确定最选择性和潜在的KGFR TKI（在特定目标＃1和2中确定的）对kGF介导的人类乳腺癌细胞在小鼠异种移植模型中的生长和转移的抑制作用。方法：在特定的目标＃1中，MCF-7乳腺癌细胞将接受刺激剂量的kgf（50 ng/ml）或以10至200？m或kgf（50 ng/ml） + kgf（50 ng/ml） + kgf tki的剂量的kgffr tki，每个抑制剂剂量或iressa均可选择的coptermal proventor tk in Compenter and coption and contrester and contrester and contrand and contrand and contrand and cottror tk Andifter and and and and and and and and and Inderive nig and Indife night in K.范围或kgf + iressa（同样的剂量）或车辆控制。在不同时间，kGF给药后的1至48小时，将收集细胞，以确定KGFR，磷酸-KGFR，ERK1/2，Phosho-erk1/2和»-Actin的蛋白质表达。在特定的视频显微镜检查中，将评估MCF-7细胞的特定目标＃2增殖和迁移，并在第一个特定目标中使用相同的实验组和剂量在1至3天内进行了1至3天的培养图测定。在特定的目标＃3中，将确定KGFR抑制对肿瘤异种移植物的生长和转移的影响。异种移植研究将通过将MCF-7细胞植入裸鼠的乳腺脂肪垫中进行。在实验结束时，将去除肿瘤，肺和肝脏，并通​​过荧光显微镜检查冻结切片，以量化肺和肝脏微碎裂的肿瘤生长和发育。意义：建立KGFR作为重要的治疗靶标，以及鉴定特定和潜在的KGFR TKI，应导致新的高度选择性治疗剂的快速发展，以防止癌症的转移性进展。建立KGFR作为一个重要的治疗靶标，再加上特定和潜在的KGFR TKI，应导致旨在防止癌症转移进展的新型高度选择性治疗剂的快速发展。该项目介绍了国家癌症研究所的几个重点领域；也就是说，癌症转移的生物学以及高度选择性和分子靶向治疗剂的发展。

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