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: 8920194
• 负责人: Lori Rink
• 金额: $ 24.15万
• 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/8920194
• 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; Commit; 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; 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; 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 伊马替尼梅赛酸盐（IM）彻底改变了胃肠道肿瘤患者的治疗 （要点）；但是，对IM的临床抵抗已经成为现实，浮力于观察到的初始效率。 此外，患者（约20％）在治疗开始时存在非常有限的选择（约20％）。使用 来自前瞻性新辅助II期试验（RTOG 0132）的临床治疗前活检样品，我确定了一个 32基因签名，包括KRAB-ZNF 91个可以预测对IM响应的亚家族成员。我 随后证明，这些基因中的许多不仅可以预测IM反应，还可以介导 该药物的活性。为了机械地确定这些ZNF如何调节对IM的响应， RNAi方法用于敲除预测特征（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分析。这导致了这个时期的发现， NEDD9和TGFβ3在siRNA介导的敲低后普遍下调。这很有趣 因为TGFβ3是阶段stin和nedd9的已知诱导剂，这两者都经常报道 过表达各种人类癌症，并表明这些ZNF从这些临床上发现 研究可能是这些途径上的“主控制器”。有趣的是，缺氧已被证明 上调NEDD9和odecin（通过PI3K）的表达，导致侵略性提高 并增强了肿瘤细胞的存活率。因此，可能会在 TGFβ和缺氧的类似机制，由ZNF控制，以调节要点细胞中的IM反应。 NEDD9还显示出过表达和SRC在不耐率的GIST细胞系中过度激活，并具有 NEDD9的敲低恢复IM敏感性。我假设要点的抗性可能是相关的 由于缺氧导致ZnF对TGFβ3进行反式激活，从而导致随后诱导时期stin和/或 NEDD9和PI3-K或SRC途径的激活。这些潜在的“救援”激活 途径”是独立于套件/pDGFRA信号传导的情况，从而克服了IM的抑制作用 生存途径。我的初步数据表明，这些ZNF负责控制此途径 这些基因的过表达与要点患者对基于IM治疗的临床抗性有关。 为了检验我的假设，我在目标1中提出建议，以确定tgfβ3，nedd9和/或ofersstin的表达是否表达 在主要的要点样品中，与IM反应（短期和长期）和ZNF表达相关。在目标2我 将研究这些ZNF如何影响TGFβ3，NEDD9和周期表达，并最终对IM响应。 AIM 3将确定靶向缺氧诱导的TGFβ3→骨膜蛋白→PI-3K/AKT和/或缺氧 - 诱导的TGFβ3→NEDD9→体内SRC救援途径将消除对IM的耐药性。中央宗旨 该提议是，当用于仅阻止的组合中时，靶向代理将变得潜力 多种致癌途径，防止平行途径的规避可以充当“救援路线” 对于这些肿瘤细胞。 拟议的研究将在宾夕法尼亚州费城的Fox Chase Cancer Center（FCCC）完成。 FCCC是由国家癌症研究所指定的综合癌症中心，全国认可 它在医学，辐射和手术肿瘤学方面的领导才能。 FCCC领导人致力于提供 必要的基础设施和支持我成功完成我建议的研究。我会被考虑 由Drs。玛格丽特·冯·梅伦（Margaret von Mehren）（肉瘤计划教授兼董事），安德鲁·K·戈德温（教授＆ 堪萨斯大学医学院（KUMC）和Erica Golemis的分子肿瘤学主任 （FCCC教授兼副首席科学官），FCCC和KUMC的尊敬的教师 国际公认的转化研究和医学肿瘤学领域的专家。我会有 在包括肉瘤在内的工作组的定期会议上进行互动和讨论我的研究的机会 参加的计划，转化研究论坛，发展治疗和双月实验室会议参加了会议 由两个Drs。冯·梅伦（Von Mehren）和戈德温（通过视频会议）。我的主要目标是获得独立 癌症研究的学术生涯。为了帮助实现这一目标，我组建了一个咨询委员会 由我的导师和其他FCCC教职员工组成（博士后主任莫琳·墨菲（Maureen Murphy）博士 培训计划和Edna Cukierman）为了审查我的研究进度并提供反馈。 在资金的头两年中，我将继续与导师紧密合作以解决研究 目标1和2中提出的目标。在此期间，我将过渡到独立性，以寻求任期 教师职位。在资金的最后三年中，我将继续在目标2和3中提出的研究。 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 仅在对新型生物制剂（例如IM）的抵抗机制中，但在建立模型和挑战时 现有的范式合理发展组合疗法以减少癌症的灼伤。 我已经在短暂的职业生涯中取得了重大的科学成就，并建立了 我自己是GIST研究社区的重要成员。我撰写了10篇研究文章， 学术评论。我已经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会议上介绍了我的工作，这是该提议的基础。