Regulation of oncogenic KIT by LMTK3

LMTK3 对致癌 KIT 的调节

• 批准号: 9260685
• 负责人: Michael C Heinrich
• 金额: --
• 依托单位: PORTLAND VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9260685
• 关键词: Acute Myelocytic Leukemia; Adult; Affect; Alleles; American; Antineoplastic Agents; Binding; Binding Proteins; Biological Assay; Biological Process; Budgets; Cancer Biology; Caring; Cell Line; Cell Survival; Cells; Clinical; Development; Disease; Drug resistance; Effectiveness; Gastrointestinal Stromal Tumors; Genetic Transcription; Growth; Healthcare; Human; Imatinib; In Vitro; Lemurs; Lesion; Libraries; Ligands; Luciferases; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Mast Cell Neoplasm; Mediating; Medical; Melanoma Cell; Messenger RNA; Methods; Morbidity - disease rate; Mutation; Nature; Neoplasms; Oncogenic; Pathogenicity; Patients; Phosphorylation Site; Phosphotransferases; Play; Protein Isoforms; Protein Tyrosine Kinase; Protein-Serine-Threonine Kinases; Proteins; Quality of life; Radiation; Receptor Protein-Tyrosine Kinases; Regulation; Reporter; Resistance; Resistance development; Role; Seminoma; Series; Small Interfering RNA; Soft tissue sarcoma; Stem Cell Factor; Toxic effect; Transcriptional Regulation; Tumor Biology; Tyrosine Kinase Inhibitor; Veterans; agent orange; cancer cell; cancer therapy; chemotherapy; clinically relevant; conventional therapy; gain of function mutation; improved; in vivo; knock-down; mRNA Expression; mast cell; melanoma; model development; mortality; mutant; neoplastic cell; novel strategies; novel therapeutic intervention; promoter; protein complex; public health relevance; resistance mutation; response; scaffold; targeted agent; targeted cancer therapy; therapeutic target; transcription factor; tumor

 DESCRIPTION (provided by applicant): Gain-of-function mutations of KIT are found in a number of human malignancies, including gastrointestinal stromal tumors (GIST), mast cell neoplasms, melanoma, seminoma, and acute myeloid leukemia. GIST tumors are resistant to both radiation and chemotherapy. KIT mutations are the oncogenic driver in the case of most GISTs. Tyrosine kinase inhibitors (TKIs) such as imatinib have revolutionized the treatment of advanced GIST. In mast cell neoplasms (SM), the disease is caused by mast cells harboring activating KIT mutations. SM patients are candidates for chemotherapy, but these therapies are toxic and ineffective. Melanomas with activating KIT mutations are aggressive and do not respond to chemotherapy. In all three diseases, the effectiveness of KIT TKIs is limited by primary or secondary drug resistance. Our proposal seeks to improve therapy for KIT-mutant malignancies by identifying the role LMTK3 has in positively regulating the expression of oncogenic KIT protein. Plan: Our proposal has three specific aims. In SA1, we will determine the mechanisms by which LMTK3 regulates the transcription of oncogenic KIT isoforms. In SA2, we will determine the functional domains of LMTK3 required for regulated expression of oncogenic KIT. In SA3, we will identify proteins that interact with LMTK3 and determine their role in promoting KIT transcription. Methods: In SA1, we will use a KIT promoter-reporter construct to identify regions of the KIT promoter that are regulated by LMTK3 knockdown. In addition, we will identify candidate transcription factors that mediate indirect regulation of KIT transcription by LMTK3. In SA2, we will create a series of LMTK3 mutants with mutation of specific domains (e.g. LMTK3 kinase domain) and determine which domains are required for regulation of the KIT promoter. In addition, we will identify LMTK3 phosphorylation sites using mass spectrometry. In SA3 we will use mass spectrometry and protein pull down assays to identify proteins that interact with LMTK3 and potentially regulate KIT transcription. Clinical Relevance: The care of veterans with cancer represents a significant portion of the overall Veterans Affairs Health Care budget. New cancer treatment agents that target significant aspects of tumor biology will greatly increase the quality of life o veterans with cancer. Our study of the mechanisms by which LMTK3 regulates oncogenic KIT proteins has the potential to identify new treatments for KIT-mutant cancers. These new treatments are likely to be more effective than conventional treatments and be associated with significantly less treatment toxicity.

 描述（由申请人提供）： 在许多人类恶性肿瘤中发现了KIT的功能获得性突变，包括胃肠道间质瘤（GIST）、肥大细胞肿瘤、黑色素瘤、腺瘤和急性髓性白血病。GIST肿瘤对放疗和化疗都有抗性。KIT突变是大多数GIST的致癌驱动因素。酪氨酸激酶抑制剂（TKI），如伊马替尼已经彻底改变了晚期GIST的治疗。在肥大细胞肿瘤（SM）中，该疾病是由携带活化KIT突变的肥大细胞引起的。SM患者是化疗的候选者，但这些疗法是有毒且无效的。具有激活KIT突变的黑色素瘤具有侵袭性，并且对化疗无反应。在所有这三种疾病中，KIT TKI的有效性受到原发性或继发性耐药性的限制。我们的建议旨在通过鉴定LMTK 3在正调节致癌KIT蛋白表达中的作用来改善KIT突变型恶性肿瘤的治疗。计划：我们的建议有三个具体目标。在SA 1中，我们将确定LMTK 3调节致癌KIT亚型转录的机制。在SA 2中，我们将确定致癌KIT的调节表达所需的LMTK 3的功能结构域。在SA 3中，我们将鉴定与LMTK 3相互作用的蛋白质，并确定它们在促进KIT转录中的作用。方法：在SA 1中，我们将使用KIT启动子-报告基因构建体来鉴定受LMTK 3敲低调控的KIT启动子区域。此外，我们将确定候选转录因子介导的间接调控KIT转录LMTK 3。在SA 2中，我们将创建一系列具有特定结构域（例如LMTK 3激酶结构域）突变的LMTK 3突变体，并确定哪些结构域是调节KIT启动子所需的。此外，我们将使用质谱法鉴定LMTK 3磷酸化位点。在SA 3中，我们将使用质谱和蛋白质下拉分析来鉴定与LMTK 3相互作用并可能调节KIT转录的蛋白质。临床相关性：癌症退伍军人的护理占退伍军人事务部整体医疗保健预算的很大一部分。针对肿瘤生物学重要方面的新癌症治疗药物将大大提高癌症退伍军人的生活质量。我们对LMTK 3调节致癌KIT蛋白的机制的研究有可能为KIT突变型癌症找到新的治疗方法。这些新的治疗方法可能比传统治疗方法更有效，并且治疗毒性显著降低。