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

• 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
• 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

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.

简介（由申请人提供）：完成医学博士学位（1999年），妇产科住院医师（1999-2003年，约翰霍普金斯大学），在NCI进行两年的基础研究培训（2003-05年），随后在克利夫兰诊所获得妇科肿瘤临床和研究奖学金（2005-08年，克利夫兰诊所），Simpkins博士被招募到迈阿密大学（UM）担任妇科肿瘤科和UM Sylvester癌症中心的助理教授和内科科学家。在她的培训期间，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，并在奖学金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的实验室指导研究，研究雌激素（E2）刺激OVCA生长的机制以及通过Src， MEK和AKT信号传导的机制。Simpkins博士拥有75%的受保护研究时间和启动资金，以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的认识，获得OVCA临床前药物开发和预测标志物的经验，精通拨款和I期试验设计。她的职业目标是开发和实施针对妇科恶性肿瘤的靶向治疗的创新策略，并在实验室和临床之间无缝移动。Simpkins博士的主要导师是J. Slingerland医学博士，他是细胞周期、类固醇激素受体生物学和激素抗性乳腺癌方面的专家。他的共同导师是M. Pegram医学博士，他是分子靶向癌症治疗的临床前和临床开发专家。癌症中心拥有一个庞大的多民族癌症患者群，拥有多学科基础科学家和临床医生的临界质量，并为临床研究提供行政支持和资源。该K08将促进Simpkins博士作为一名在分子机制研究方面受过训练的临床医生科学家的发展，其目标是产生新的治疗方法并在临床试验中进行测试。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是无法治愈的，目标是缓解症状。患者用尽了治疗方案，因此迫切需要新的治疗方法。抗雌激素疗法耐受性良好，因此在这种情况下很有吸引力。这种疗法尚未在早期（IC/II） ovca中进行测试，也许在这种情况下也有好处。该项目将有可能为OVCA提供新的治疗方案。E2在体外和体内驱动OVCA生长12-15。E2剥夺和雌激素受体（ER）阻断会导致乳腺癌（BRCA）细胞在细胞周期的G1期阻滞16,17。细胞周期进程是由细胞周期蛋白依赖性激酶（cdks）控制的，cdks被细胞周期蛋白激活，并被cdk抑制剂抑制。cdk抑制剂p21和p27是周期阴性调节因子，在包括OVCAs16、17在内的许多癌症中经常减少。ER在大多数OVCAs中表达，类似于BRCA18。抗雌激素对OVCAs存活和细胞周期的影响尚未明确。尽管在OVCAs中ER表达较高，但抗雌激素疗法的小型试验结果令人失望。抗雌激素的益处尚未在大型精心设计的OVCA试验中得到研究。在ovca中，许多患者表现出对抗雌激素的从头耐药，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ovcas1中构成性激活我的初步数据和其他人的数据支持这样一种观点，即雌激素结合的内质网和信号激酶之间的串扰导致Src和MEK/MAPK活化，从而刺激细胞周期进程，17,34。我们的初步数据显示，E2刺激ER+ OVCA细胞系中Src、MEK和AKT的进一步激活。Src抑制剂联合ER阻断通过增加p27的水平和作用引起ER+ OVCA细胞更大的G1阻滞。此外，使用Src和MEK抑制剂药物比单一治疗更有效地阻滞G1期细胞，抑制Src、MAPK和AKT，增加p27水平，抑制细胞周期蛋白E-Cdk2。虽然大多数ovca表达ER，但抗雌激素一直令人失望。联合使用抗雌激素和阻断关键有丝分裂信号通路的药物可以预防或延缓ER+ OVCAs抗雌激素耐药的发展。联合使用激酶抑制剂阻断不同的组成激活途径也可能更有效地抑制OVCA细胞的生长。我们的假设是Src和/或Ras/Raf/MEK的组成性激活促进细胞周期进程，使ER+ OVCA细胞具有抗雌激素抗性。我们进一步假设Src和MEK抑制剂可能共同或与抗雌激素合作逆转耐药并减少OVCA细胞增殖。AIM 1将在一系列扩大的ER阳性OVCA细胞系中测试不同Src抑制剂-药物组合的抗增殖活性，并研究细胞周期阻滞和凋亡的机制。AIM 2将比较药物对减体积手术切除的早期和晚期ovca直接衍生的原代培养物的影响。AIM 3将使用一种创新的小鼠OVCA肾胶囊模型，测试Src联合抗雌激素或激酶抑制剂对异种移植物的抗肿瘤效果。临床前数据将有助于确定具有最大抗肿瘤疗效的新药物组合，并可在未来的临床试验中进行测试。虽然在机制上不是新的，但靶向ER和激活通路对于迫切需要新疗法的复发性OVCA患者来说是一种新的治疗机会。抗雌激素治疗早期OVCA可能被证明可以预防或延迟复发，但从未被探索过。