Targeting SRC Signaling Pathways to Promote Cell Cycle Arrest in Ovarian Cancer

靶向 SRC 信号通路促进卵巢癌细胞周期停滞

基本信息

批准号：
8808036
负责人：
FIONA SIMPKINS
金额：
$ 16.15万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8808036
关键词：
Adjuvant Chemotherapy American Association of Cancer Research American Society of Clinical Oncology Apoptosis Aromatase Inhibitors BRAF gene Basic Science Binding Biology Breast Cancer Cell Cancer Cell Growth Cancer Center Cancer Patient Cancer cell line Cell Cycle Cell Cycle Arrest Cell Cycle Progression Cell Proliferation Cell Survival Cells Clinic Clinical Clinical Data Clinical Drug Development Clinical Research Clinical Trials Clinical Trials Design Cyclin E Cyclin-Dependent Kinases Cyclins Data Development Diagnosis Diagnostic Neoplasm Staging Doctor of Philosophy Drug Combinations Early treatment Educational workshop Estrogen Antagonists Estrogen Receptor Modulators Estrogen Receptors Estrogen Therapy Estrogen receptor positive Estrogens Etiology Exhibits Fellowship Fibrous capsule of kidney Fostering Fulvestrant Funding Future G1 Arrest G1 Phase Gene Expression Profile Goals Grant Growth Growth Factor Gynecologic Oncology Hormones Human In Vitro KRAS2 gene Knowledge Letrozole MAP Kinase Gene MEKs Malignant Neoplasms Malignant neoplasm of ovary Mediating Mentors Modeling Molecular Molecular Target Mus Mutation Neoplasm Metastasis Operative Surgical Procedures Ovarian PI3K/AKT PIK3CA gene PTEN gene Pathway interactions Patients Peripheral Pharmaceutical Preparations Pharmacotherapy Phase Phase I Clinical Trials Phosphotransferases Physicians Protein Tyrosine Kinase Proto-Oncogene Proteins c-akt Ras/Raf Receptor Activation Receptor Protein-Tyrosine Kinases Receptor Signaling Recruitment Activity Recurrence Recurrent disease Regimen Relapse Research Research Training Residencies Resistance Resistance development Resources Scientist Series Signal Pathway Signal Transduction Staging Steroids Symptoms Testing Time Toxic effect Training Tumor Debulking Universities Work Xenograft procedure base cancer cell cancer therapy career chemotherapy deprivation design exhaust experience hormone therapy human FRAP1 protein improved in vivo inhibitor/antagonist innovation kinase inhibitor malignant breast neoplasm novel novel therapeutics overexpression palliation pre-clinical prevent professor receptor expression response src Genes steroid hormone receptor tumor tumor progression tyrosine receptor

项目摘要

DESCRIPTION (provided by applicant): On completion of her MD (1999), Residency in OB-GYN (1999-2003, Johns Hopkins), two yrs of basic research training at the NCI (2003-05) followed by a Clinical and Research Fellowship in Gynecologic Oncology (2005-08, Cleveland Clinic), Dr. Simpkins was recruited to the University of Miami (UM) as Assistant Professor and physician scientist in the Division of Gynecologic Oncology and UM Sylvester Cancer Center. During her training, she completed 3 yrs of lab research, attended the AACR/ASCO Clinical Trial Design Workshop, and designed and executed a phase II ovarian cancer (OVCA) trial during fellowship. She began mentored research in Aug, 2008 in Slingerland's lab, investigating mechanisms of growth stimulation in OVCA by estrogens (E2) and signaling via Src, MEK and AKT. Dr. Simpkins has 75% protected research time and start-up funds to support initial project development to garner independent funding. The applicant's goals for the next 2-3 yrs are to deepen her knowledge of estrogen and growth factor signaling in OVCA, gain experience with pre-clinical drug development and predictive markers in OVCA, become proficient in grantsmanship and phase I trial design. Her career goal is to develop and implement innovative strategies in targeted therapy for Gyn malignancies and move seamlessly between lab and clinic. Dr. Simpkins has J. Slingerland MD, PhD as primary mentor, an expert in cell cycle, steroid hormone receptor biology and hormone resistant breast cancer and M. Pegram MD, an expert in pre-clinical and clinical development of molecular targeted cancer therapies, as co-mentor. The Cancer Center has a large multi- ethnic cancer patient base, a critical mass of multi-disciplinary basic scientists and clinicians with a translational track record, and offers admininistrative support and resources for clinical research. This K08 will foster Dr. Simpkins' development as a clinician scientist trained in molecular mechanistic research whose goal is to generate novel therapies and test them in clinical trials. A detailed training plan is proposed. OVCA is the most lethal gynecological malignancy. Most patients are diagnosed with advanced stage III/IV and recur within 2 yrs. Recurrent OVCA is incurable and goals are palliation of symptoms. Patients exhaust treatment options so new therapies are desperately needed. Anti-estrogen therapies are well tolerated and thus appealing in this setting. Such therapies have not yet been tested in early stage (IC/II) OVCAs and maybe of benefit in this setting as well. This project will potentially identify novel treatment options for OVCA. E2 drives OVCA growth in vitro and invivo12-15. E2 deprivation and estrogen receptor(ER) blockade because breast cancer (BRCA) cells to arrest in the G1 phase of the cell cycle16, 17. Cell cycle progression is governed by cyclin-dependent kinases (cdks) that are activated by cyclins and inhibited by cdk inhibitors. The cdk inhibitors, p21 and p27, are cycle negative regulators that are frequently reduced in many cancers, including OVCAs16, 17. The ER is expressed in the majority of OVCAs, similar to BRCA18. Anti-estrogen effects on survival and cell cycle in OVCAs have not been characterized. Despite high ER expression in OVCAs, small trials of anti-estrogen therapies have been disappointing. The benefit of anti-estrogens has not been studied in large well-designed OVCA trials. In OVCAs, many patients manifest de novo resistance to anti-estrogens and those that do initially respond, invariably develop resistance. Src, Ras/Raf/MEK and PI3K pathways are constitutively activated in OVCAs.1 My preliminary data and that of others support the notion that cross talk between estrogen-bound ER and signaling kinases leads to Src and MEK/MAPK activation to stimulate cell cycle progression, 17, 34. Our preliminary data show that E2 stimulates further activation of Src, MEK and AKT in ER+ OVCA lines. Combined Src inhibitor with ER blockade caused a greater G1 arrest of ER+ OVCA cells through increased levels and action of p27. Moreover, use of both Src and MEK inhibitor drugs more effectively arrested cells in G1 than monotherapy, with inhibition of Src, MAPK and AKT and increased p27 levels causing inhibition of cyclin E-Cdk2. While most OVCAs express ER, anti- estrogens have been disappointing. Combined use of antiestrogens with drugs that block critical mitogenic signaling pathways may prevent or delay development of anti-estrogen resistance in ER+ OVCAs.The combined use of kinase inhibitors that block different constitutively activated pathways may also more effectively inhibit OVCA cell growth. Our HYPOTHESIS is that constitutive activation of Src and/or Ras/Raf/MEK promote cell cycle progression, rendering ER+ OVCA cells anti-estrogen resistant. We further postulate that Src and MEK inhibitors may cooperate together or with anti-estrogens to reverse resistance and decrease OVCA cell proliferation. AIM 1 will test different Src inhibitor-drug combinations for anti-proliferative activity in an expanded series of ER positive OVCA cell lines and study the mechanisms of cell cycle arrest and apoptosis. AIM 2 will compare drug effects on primary cultures derived directly from early and advanced stage OVCAs removed at debulking surgery. AIM 3 will test the anti-tumor efficacy of combined Src inhibition with an antiestrogen or kinase inhibitor on xenografts using an innovative mouse OVCA renal capsule model. The pre-clinical data will help to identify novel drug combinations with the greatest anti-tumor efficacy that can be tested in future clinical trial. Although not mechanistically new, targeting ER and activated pathways is a novel therapeutic opportunity for recurrent OVCA patients who desperately need new therapies. Anti-estrogen treatment of early stage OVCA may prove to prevent or delay recurrence, which has never been explored.

描述（由申请人提供）：完成医学博士学位（1999 年）、妇产科住院医师实习（1999-2003 年，约翰·霍普金斯大学）、在 NCI 接受两年基础研究培训（2003-05 年），随后进行临床和研究培训妇科肿瘤学研究员（2005-08，克利夫兰诊所），Simpkins 博士被招募到迈阿密大学（密歇根大学）妇科肿瘤科和密歇根大学西尔维斯特癌症中心的助理教授和医师科学家。在培训期间，她完成了 3 年的实验室研究，参加了 AACR/ASCO 临床试验设计研讨会，并在进修期间设计并执行了一项 II 期卵巢癌 (OVCA) 试验。她于 2008 年 8 月开始在 Slingerland 实验室指导研究，研究雌激素 (E2) 刺激 OVCA 生长的机制以及通过 Src、MEK 和 AKT 发出的信号。 Simpkins 博士拥有 75% 受保护的研究时间和启动资金，用于支持初始项目开发以获得独立资金。申请人未来2-3年的目标是加深对OVCA中雌激素和生长因子信号传导的了解，获得OVCA临床前药物开发和预测标记的经验，精通资助和I期试验设计。她的职业目标是开发和实施妇科恶性肿瘤靶向治疗的创新策略，并在实验室和诊所之间无缝转移。 Simpkins 博士的主要导师是细胞周期、类固醇激素受体生物学和激素抵抗性乳腺癌方面的专家 J. Slingerland MD 博士，以及分子靶向癌症疗法临床前和临床开发方面的专家 M. Pegram MD。作为共同导师。癌症中心拥有庞大的多种族癌症患者群体、大量具有转化记录的多学科基础科学家和临床医生，并为临床研究提供行政支持和资源。 K08 将促进 Simpkins 博士作为一名接受过分子机制研究培训的临床科学家的发展，其目标是产生新的疗法并在临床试验中对其进行测试。提出了详细的培训计划。 OVCA 是最致命的妇科恶性肿瘤。大多数患者被诊断为晚期 III/IV 期，并在 2 年内复发。复发性 OVCA 无法治愈，目标是缓解症状。患者已经耗尽了治疗选择，因此迫切需要新的疗法。抗雌激素疗法具有良好的耐受性，因此在这种情况下很有吸引力。此类疗法尚未在早期 (IC/II) OVCA 中进行测试，并且可能在这种情况下也有好处。该项目将有可能确定 OVCA 的新治疗方案。 E2 在体外和体内促进 OVCA 生长12-15。 E2 剥夺和雌激素受体 (ER) 阻断，因为乳腺癌 (BRCA) 细胞停滞在细胞周期的 G1 期16, 17。细胞周期进展受细胞周期蛋白依赖性激酶 (cdks) 控制，该激酶被细胞周期蛋白激活并被细胞周期蛋白抑制cdk抑制剂。 cdk 抑制剂 p21 和 p27 是周期负调节因子，在许多癌症中经常减少，包括 OVCAs16、17。ER 在大多数 OVCA 中表达，与 BRCA18 类似。抗雌激素对 OVCA 存活和细胞周期的影响尚未得到表征。尽管 OVCA 中 ER 表达较高，但抗雌激素疗法的小规模试验却令人失望。大型精心设计的 OVCA 试验尚未研究抗雌激素的益处。在 OVCA 中，许多患者表现出对抗雌激素的新耐药性，而那些最初有反应的患者，总是会产生耐药性。 Src、Ras/Raf/MEK 和 PI3K 通路在 OVCA 中被组成型激活。1 我和其他人的初步数据支持这样的观点：雌激素结合的 ER 和信号激酶之间的串扰导致 Src 和 MEK/MAPK 激活，从而刺激细胞周期进展, 17, 34。我们的初步数据表明，E2 刺激 ER+ OVCA 系中 Src、MEK 和 AKT 的进一步激活。 Src 抑制剂与 ER 阻断相结合，通过增加 p27 的水平和作用，引起 ER+ OVCA 细胞更大的 G1 期停滞。此外，使用 Src 和 MEK 抑制剂药物比单一疗法更有效地将细胞阻滞在 G1 期，抑制 Src、MAPK 和 AKT，并增加 p27 水平，从而抑制细胞周期蛋白 E-Cdk2。虽然大多数 OVCA 表达 ER，但抗雌激素却令人失望。联合使用抗雌激素与阻断关键有丝分裂信号通路的药物可能会预防或延缓 ER+ OVCA 中抗雌激素耐药性的发展。联合使用阻断不同组成型激活通路的激酶抑制剂也可能更有效地抑制 OVCA 细胞生长。我们的假设是，Src 和/或 Ras/Raf/MEK 的组成型激活促进细胞周期进展，使 ER+ OVCA 细胞产生抗雌激素抵抗。我们进一步假设 Src 和 MEK 抑制剂可能共同作用或与抗雌激素协同作用以逆转耐药性并减少 OVCA 细胞增殖。 AIM 1 将测试不同的 Src 抑制剂-药物组合在一系列扩展的 ER 阳性 OVCA 细胞系中的抗增殖活性，并研究细胞周期停滞和细胞凋亡的机制。 AIM 2 将比较药物对直接来自减灭手术中取出的早期和晚期 OVCA 的原代培养物的影响。 AIM 3 将使用创新的小鼠 OVCA 肾囊模型来测试 Src 抑制与抗雌激素或激酶抑制剂联合对异种移植物的抗肿瘤功效。临床前数据将有助于确定具有最大抗肿瘤功效的新型药物组合，并可在未来的临床试验中进行测试。虽然从机制上来说并不新鲜，但针对急需新疗法的复发性 OVCA 患者来说，针对 ER 和激活途径是一个新的治疗机会。早期 OVCA 的抗雌激素治疗可能被证明可以预防或延迟复发，但这一点从未被探索过。