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：
8472456
负责人：
FIONA SIMPKINS
金额：
$ 16.15万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
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 2013-12-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8472456
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）：在完成MD（1999年），Ob-Gyn的居住期（1999-2003，Johns Hopkins），NCI（2003-05）的基础研究培训的两年年限（2003-05），然后是临床和研究的临床和研究，随后是妇科学的临床和研究。妇科肿瘤学和UM Sylvester癌症中心的医师科学家。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，并在奖学金期间设计和执行了II期卵巢癌（OVCA）试验。她于2008年8月在Slingerland的实验室开始指导研究，研究了雌激素（E2）（E2）和通过SRC，MEK和AKT发出信号的OVCA生长刺激机制。 Simpkins博士拥有75％的保护时间和启动资金，以支持Garner Independent资助的初始项目开发。接下来的2 - 3年的申请人目标是加深她对OVCA中雌激素和生长因子信号的了解，在临床前药物开发方面获得经验以及OVCA的预测标记，熟练精通赠款和I期试验设计。她的职业目标是制定和实施针对Gyn恶性肿瘤的有针对性疗法的创新策略，并在实验室和诊所之间无缝移动。 Simpkins博士拥有J. Slingerland MD博士学位，是主要指导者，是细胞周期专家，类固醇激素受体生物学和耐激素乳腺癌和M. Pegram M. Pegram M.，M. Pegram MD是分子靶向癌症疗法的临床前和临床发育专家，作为共同群体。癌症中心拥有大量的多民族癌症患者基础，具有转化往绩的多学科基础科学家和临床医生的临界量，并为临床研究提供了管理的支持和资源。该K08将促进辛普金斯博士作为一名临床科学家的发展，接受了分子机械研究培训的临床科学家，其目标是在临床试验中产生新颖的疗法并对其进行测试。提出了详细的培训计划。 OVCA是最致命的妇科恶性肿瘤。大多数患者被诊断出患有晚期III/IV期，并在2年内复发。复发性OVCA是无法治愈的，目标是症状的抚慰。患者排气治疗选择，因此迫切需要新的疗法。抗雌激素疗法的耐受性很好，因此在这种情况下具有吸引力。此类疗法尚未在早期（IC/II）卵子进行测试，在这种情况下也可能有益。该项目将有可能确定OVCA的新型治疗选择。 E2在体外和Invivo12-15中驱动OVCA增长。 E2剥夺和雌激素受体（ER）阻滞是因为乳腺癌（BRCA）细胞在细胞周期的G1阶段中停滞17。细胞周期进展受被细胞周期蛋白激活并抑制CDK抑制剂抑制的细胞周期蛋白依赖性激酶（CDKS）。 CDK抑制剂P21和P27是循环负调节剂，在包括OVCAS16，17在内的许多癌症中经常会降低。ER在大多数OVCA中表达，类似于BRCA18。尚未表征抗雌激素对卵子生存和细胞周期的影响。尽管在卵子中表达高ER，但抗雌激素疗法的小试验令人失望。在大型精心设计的OVCA试验中尚未研究抗雌激素的好处。在OVCA中，许多患者表现出对抗雌激素的抵抗力，而最初反应的患者总是会产生抗药性。 SRC，RAS/RAF/MEK和PI3K途径在OVCAS中受到组成激活。1我的初步数据，而其他数据则支持以下观点，即雌激素结合ER之间的交叉交流和信号激酶之间的交叉交流导致SRC和MEK/MAPK激活，以刺激细胞周期，我们的PRELERIMIAL AT。 ER+ OVCA线。通过p27的水平和作用增加，SRC抑制剂与ER封闭的结合抑制剂与ER+ OVCA细胞的停滞更大。此外，与单一疗法相比，使用SRC和MEK抑制剂药物在G1中更有效地阻止细胞，并抑制SRC，MAPK和AKT，并增加了P27水平，从而导致Cyclin e-CDK2抑制。虽然大多数卵子都表达，但抗浓缩者一直令人失望。抗雌激素与阻断关键有丝分裂信号通路的药物的联合使用可能会阻止或延迟ER+ OVCAS中抗雌激素耐药性的发展。激酶抑制剂的联合使用可以更有效地抑制OVCA细胞的生长。我们的假设是SRC和/或RAS/RAF/MEK的组成型激活促进了细胞周期的进程，从而使ER+ OVCA细胞具有抗雌激素的抗基因。我们进一步假设SRC和MEK抑制剂可以合作或与抗雌激素合作以逆转耐药性并减少OVCA细胞增殖。 AIM 1将在一系列ER阳性OVCA细胞系中测试不同的SRC抑制剂 - 药物组合，以实现抗增殖活性，并研究细胞周期停滞和凋亡的机制。 AIM 2将比较直接从早期和晚期的卵子中衍生出的药物影响，该培养物在脱离手术中去除。 AIM 3将使用创新的小鼠OVCA肾胶囊模型在异种移植物上使用抗雌激素或激酶抑制剂联合抑制的抗肿瘤功效。临床前数据将有助于确定可以在以后的临床试验中测试的最大抗肿瘤功效的新型药物组合。尽管不是机械上的新事物，但针对ER和激活途径是急需新疗法的复发性OVCA患者的新型治疗机会。早期OVCA的抗雌激素治疗可能证明是预防或延迟复发，这从未探索过。