ZNF-Mediated Resistance to Imatinib Mesylate in Gastrointestinal Stromal Tumor

ZNF 介导的胃肠道间质瘤对甲磺酸伊马替尼的耐药性

• 批准号: 8928568
• 负责人: Lori Rink
• 金额: $ 24.9万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8928568
• 关键词: 70-kDa Ribosomal Protein S6 Kinases; Address; Adult; Advisory Committees; Affect; American Association of Cancer Research; American Society of Clinical Oncology; Award; Biological Assay; Biological Markers; Biopsy Specimen; Cancer Burden; Cell Line; Cells; Clinic; Clinical; Clinical Research; Combined Modality Therapy; Communities; Comprehensive Cancer Center; Dasatinib; Data; Developmental Therapeutics Program; Drug resistance; Exons; Faculty; Feedback; Fellowship; Fox Chase Cancer Center; Funding; Gastrointestinal Stromal Tumors; Genes; Goals; Human; Hypoxia; Imatinib mesylate; Immunoblotting; In Vitro; Kansas; Leadership; Light; Malignant Neoplasms; Measures; Mediating; Medical Oncology; Mentors; Modeling; Molecular Profiling; National Cancer Institute; Neoadjuvant Therapy; Oncogenic; PDGFRA gene; PI3K/AKT; Pathway interactions; Patients; Pharmaceutical Preparations; Phase II Clinical Trials; Philadelphia; Positioning Attribute; Proto-Oncogene Proteins c-akt; Public Health; RNA Interference; RNA Splicing; Radiation Oncology; Radiation Therapy Oncology Group; Recurrence; Refractory; Reporting; Research; Research Infrastructure; Resistance; Role; Route; Sampling; Scientific Advances and Accomplishments; Serum; Signal Pathway; Signal Transduction; Small Interfering RNA; Stream; Surgical Oncology; Testing; Time; Training; Training Programs; Transactivation; Transforming Growth Factor beta; Translating; Translational Research; Tumor Cell Line; Universities; Videoconferences; Videoconferencing; Work; Xenograft Model; anticancer research; base; career; cytotoxicity; experience; human ZNF45 protein; improved; in vivo; inhibitor/antagonist; interest; medical schools; meetings; member; molecular oncology; neoplastic cell; novel; novel strategies; overexpression; periostin; phase II trial; post-doctoral training; pre-clinical; prevent; professor; programs; prospective; research study; resistance mechanism; response; sarcoma; small molecule; sunitinib malate; tumor; tumor growth; tumor xenograft; tumorigenesis; working group

PROJECT SUMMARY/ABSTRACT Imatinib mesylate (IM) has revolutionized the treatment of patients with gastrointestinal stromal tumors (GISTs); however, clinical resistance to IM has become a reality, despite the initial efficacy observed. Furthermore, very limited options exist for patients (~20%) that are refractory at the start of treatment. Using clinical pre-treatment biopsy samples from a prospective neoadjuvant phase II trial (RTOG 0132), I identified a 32-gene signature that includes KRAB-ZNF 91 subfamily members that can predict response to IM. I subsequently demonstrated that many of these genes were not only predictive of IM response but mediated the drug's activity. In order to determine mechanistically how these ZNFs might be modulating response to IM, RNAi approaches were used to knockdown genes within the predictive signature (including 10 ZNFs) in GIST cells and expression profiling was performed using exon 1.0 ST arrays. This led to the finding that periostin, NEDD9 and TGFβ3, are universally downregulated following siRNA-mediated knockdown. This is intriguing because TGFβ3 is a known inducer of periostin and NEDD9, both of which have been reported as frequently overexpressed in a variety of human cancers and indicates that these ZNFs uncovered from these clinical studies may be a “master controller” over these pathways. Interestingly, hypoxia has been shown to upregulate expression of both NEDD9 and periostin (through the PI3K) leading to increased aggressiveness and enhanced survival of tumor cells. Therefore, it is possible that both periostin and NEDD9 are regulated in a similar mechanism by TGFβ and hypoxia, controlled by ZNFs, to modulate IM response in GIST cells. NEDD9 was also shown to be overexpressed and SRC hyper-activated in an IM-resistant GIST cell line, with knockdown of NEDD9 restoring IM sensitivity. I hypothesize that IM resistance in GISTs may be associated with hypoxia leading to the transactivation of TGFβ3 by ZNFs causing subsequent induction of periostin and/or NEDD9 and activation of the PI3-K or SRC pathways, respectively. The activation of these potential “rescue routes” occurs independent of KIT/PDGFRA signaling, thereby overcoming inhibiting activities of IM on pro- survival pathways. My preliminary data suggest that these ZNFs are responsible for regulating this pathway and overexpression of these genes is associated with clinical resistance of GIST patients to IM-based therapy. To test my hypothesis, I propose in Aim 1 to determine if expression of TGFβ3, NEDD9 and/or periostin in primary GIST samples correlate to IM response (both short and long-term) and ZNF expression. In Aim 2 I will investigate how these ZNFs affect TGFβ3, NEDD9 and periostin expression and ultimately response to IM. Aim 3 will establish whether targeting the hypoxia-induced TGFβ3→ periostin→ PI-3K/AKT and/or hypoxia- induced TGFβ3→ NEDD9→ SRC rescue pathway in vivo will abrogate resistance to IM. The central tenet of this proposal is that targeted agents will become potent when used in combinations that simultaneously block multiple oncogenic pathways, preventing the circumvention of parallel pathways that can act as “rescue routes” for these tumor cells. The proposed research will be accomplished at Fox Chase Cancer Center (FCCC) in Philadelphia, PA. FCCC is a comprehensive cancer center as designated by the National Cancer Institute, nationally recognized for its leadership in medical, radiation and surgical oncology. FCCC leadership is committed to providing the necessary infrastructure and support for me to successfully complete my proposed studies. I will be mentored by Drs. Margaret von Mehren (Professor & Director, Sarcoma Program), Andrew K. Godwin (Professor & Director of Molecular Oncology at the University of Kansas Medical School (KUMC), and Erica Golemis (Professor & Deputy Chief Scientific Officer at FCCC), esteemed faculty members at FCCC and KUMC and internationally recognized experts in the field of translational research and medical oncology. I will have the opportunity to interact and discuss my research in regular meetings of working groups including the Sarcoma Program, Translational Research Forum, Developmental Therapeutics and bi-monthly lab meetings attended by both Drs. von Mehren and Godwin (by videoconference). My primary goal is to obtain an independent academic career in cancer research. To help accomplish this goal, I have assembled an advisory committee consisting of my mentors and other FCCC faculty members (Drs. Maureen Murphy, Director of Postdoctoral Training Program and Edna Cukierman) in order to review my research progress and provide feedback. During the first two years of funding, I will continue to work closely with my mentors to address the research goals proposed in Aims 1 and 2. During this time I will transition into independence seeking out a tenure-track faculty position. In the last three years of funding, I will continue the studies proposed in Aims 2 and 3 while establishing my independent research program. As part of my future research objectives, I am interested not only in mechanisms of resistance to novel biologics, such as IM, but in establishing models and challenging existing paradigms to rationally develop combination therapies in order to reduce the burden of cancer. I have already made significant scientific accomplishments in my short career and have established myself to be an important member of the GIST research community. I have authored 10 research articles and scholarly reviews. I have received a number of awards and training fellowships and was selected to orally present my work, the basis of this proposal, at both the AACR and ASCO meetings in 2009.

项目摘要/摘要 甲磺酸伊马替尼(IM)使胃肠道间质瘤患者的治疗发生了革命性变化 然而，尽管观察到最初的疗效，临床上对IM的耐药性已经成为现实。 此外，对于在治疗开始时难治的患者(~20%)来说，选择非常有限。vbl.使用 临床治疗前活检样本来自一项预期的新辅助II期试验(RTOG 0132)，I确定了一种 32基因签名，包括KRAB-ZNF91亚家族成员，可预测对IM的反应。我 随后证明，这些基因中的许多不仅是IM反应的预测，而且是中介的 药物的活性。为了机械地确定这些ZNF可能如何调制对IM的响应， 在GIST中使用RNAi方法敲除预测信号(包括10个ZNF)内的基因 使用外显子1.0 ST阵列进行细胞和表达谱分析。这导致了Periostin的发现， 在小干扰RNA介导的基因敲除后，NEDD9和转化生长因子β3普遍下调。这很耐人寻味 因为转化生长因子β3是已知的Periostin和NEDD9的诱导剂，这两种物质都被频繁报道 在多种人类癌症中过表达，并表明这些ZNF在这些临床上发现 研究可能是这些途径的“主控者”。有趣的是，低氧已被证明 上调NEDD9和Periostin的表达(通过PI3K)导致侵袭性增加 并提高肿瘤细胞的存活率。因此，Periostin和NEDD9都有可能在 转化生长因子β和低氧调节胃肠间质瘤细胞IM反应的机制类似。 NEDD9也被证明在IM耐药的GIST细胞系中过表达和SRC高激活， 击倒NEDD9，恢复即时通讯的敏感性。我推测GIST中的IM抵抗可能与 低氧导致ZNF反式激活转化生长因子β3，从而诱导Periostin和/或 NEDD9和PI3-K或SRC通路的激活。这些潜在的“救援”被激活 途径“独立于KIT/PDGFRA信号发生，从而克服IM对PRO的抑制活性. 生存之路。我的初步数据表明，这些零核因子负责调节这一途径 这些基因的过度表达与GIST患者对IM治疗的临床抵抗力有关。 为了验证我的假设，我在目标1中提出了确定转化生长因子β3、NEDD9和/或Periostin的表达 在初级GIST样本中，IM反应(包括短期和长期)和ZNF表达相关。在AIM 2中I 将研究这些ZNF如何影响转化生长因子β3、NEDD9和Periostin的表达，并最终影响IM的反应。 目的3将确定靶向低氧诱导的转化生长因子β3→Periostin→PI-3K/AKT和/或低氧- 体内诱导的转化生长因子-β-3-→-NEDD9-→-SRC救援通路可消除对IM的耐药性。的核心信条 这一提议是，靶向药物在联合使用时将变得有效，同时阻止 多条致癌途径，防止绕过可充当“救援途径”的平行途径 这些肿瘤细胞。 这项拟议的研究将在宾夕法尼亚州费城的福克斯·蔡斯癌症中心完成。 Fccc是由国家癌症研究所指定的综合性癌症中心，国家认可。 以表彰其在内科、放射和外科肿瘤学领域的领导地位。Fccc领导层致力于提供 我需要必要的基础设施和支持，才能顺利完成我提议的学业。我会得到指导的 作者：玛格丽特·冯·迈伦博士(肉瘤项目教授兼主任)，安德鲁·K·戈德温(教授& 堪萨斯大学医学院分子肿瘤学主任和埃里卡·戈莱米斯 (fccc教授兼副首席科学官)，fccc和kumc尊敬的教职员工，以及 翻译研究和医学肿瘤学领域的国际公认专家。我要一份 有机会在包括肉瘤在内的工作组的定期会议上互动和讨论我的研究 参加计划、转化研究论坛、发展治疗和双月一次的实验室会议 Von Mehren博士和Godwin博士(通过视频会议)。我的主要目标是获得一个独立的 从事癌症研究的学术生涯。为了帮助实现这一目标，我成立了一个咨询委员会。 由我的导师和fccc的其他教职员工组成(博士后主任莫琳·墨菲博士 培训计划和Edna Cukierman)，以便审查我的研究进展并提供反馈。 在资助的头两年里，我将继续与我的导师们密切合作，解决研究问题 目标1和目标2中提出的目标。在此期间，我将过渡到独立，寻找终身教职的轨道 教职员工职位。在最后三年的资助中，我将继续进行目标2和目标3中提出的研究，同时 建立我的独立研究计划。作为我未来研究目标的一部分，我不感兴趣 仅限于对新生物制剂(如IM)的抵抗机制，但在建立模型和挑战方面 合理开发联合疗法以减轻癌症负担的现有范例。 在我短暂的职业生涯中，我已经取得了重大的科学成就，并建立了 我想成为GIST研究界的重要一员。我写了10篇研究文章， 学术评论。我获得了许多奖项和培训奖学金，并被选为口头 在2009年的AACR和ASCO会议上介绍我的工作，这是这项提议的基础。