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: 8531192
• 负责人: Lori Rink
• 金额: $ 10.1万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8531192
• 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; 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; public health relevance; research study; resistance mechanism; response; sarcoma; small molecule; sunitinib malate; tumor; tumor growth; tumor xenograft; tumorigenesis; working group

DESCRIPTION (provided by applicant): 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 TGF23, are universally downregulated following siRNA-mediated knockdown. This is intriguing because TGF23 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 TGF2 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 TGF23 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 TGF23, 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 TGF23, NEDD9 and periostin expression and ultimately response to IM. Aim 3 will establish whether targeting the hypoxia-induced TGF23-> periostin-> PI-3K/AKT and/or hypoxia- induced TGF23-> 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 wil 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.

描述（由申请人提供）：伊马替尼麦甲酸酯（IM）彻底改变了胃肠道肿瘤患者的治疗（GISTS）；但是，尽管观察到了最初的功效，但对IM的临床抵抗已成为现实。此外，患者（约20％）在治疗开始时存在非常有限的选择（约20％）。使用临床前 我从前瞻性新辅助II期试验（RTOG 0132）的治疗活检样品中确定了一个32基因的特征，其中包括KRAB-ZNF 91个下属成员，可以预测可以预测对IM的响应。随后，我证明了其中许多基因不仅可以预测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，使用RNAi方法来敲除GIST细胞中预测性特征（包括10个ZNF）内的基因，并使用外显子1.0 ST阵列进行了表达分析。这导致发现，骨膜介导的敲低后，骨膜蛋白NEDD9和TGF23被普遍下调。这很有趣，因为TGF23是骨膜蛋白和NEDD9的已知诱导剂，这两者都在各种人类癌症中经常过表达，并表明从这些临床研究中发现的这些ZNF可能是这些途径上的“主控制器”。有趣的是，已经显示缺氧可以上调NEDD9和骨膜素（通过PI3K）的表达，从而增加侵袭性和增强肿瘤细胞存活率。因此，骨膜蛋白和NEDD9可能都受到ZNF控制的TGF2和缺氧的类似机制调节，以调节GIST细胞中的IM反应。 NEDD9还显示出过表达和SRC在不抗性的GIST细胞系中过度激活，而NEDD9的敲低恢复了IM敏感性。我假设GIST的IM耐药性可能与缺氧有关，从而导致ZnFS导致TGF23的缺氧，从而导致随后诱导骨膜蛋白和/或NEDD9，并分别激活PI3-K或SRC途径。这些潜在的“救援途径”的激活独立于KIT/PDGFRA信号传导，从而克服了IM对生存途径的抑制活性。我的初步数据表明，这些ZNF负责调节这种途径，这些基因的过表达与GIST患者对基于IM的治疗的临床抗性有关。 为了检验我的假设，我在AIM 1中提出建议，以确定在原发性GIST样品中TGF23，NEDD9和/或骨膜素的表达是否与IM响应（短期和长期）和ZNF表达相关。在AIM 2中，我将研究这些ZNF如何影响TGF23，NEDD9和Pererostin表达，并最终对IM响应。 AIM 3将确定靶向缺氧诱导的TGF23-> Pererostin-> perostin-> pi-3K/Akt和/或缺氧诱导的TGF23-> NEDD9-> src Rescue途径在体内将消除对IM的抵抗力。该提案的中心宗旨是，当使用同时阻止多个致癌途径的组合中，靶向剂将变得有效，从而阻止了平行途径的规避，这些途径可以用作这些肿瘤细胞的“救援路线”。 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，该中心因其在医疗，辐射和手术肿瘤学领域的领导而获得全国认可。 FCCC领导层致力于为我提供必要的基础设施和支持，以成功完成我的拟议研究。我将受到博士的指导。玛格丽特·冯·梅伦（Margaret von Mehren）（肉瘤计划教授兼主任），安德鲁·K·戈德温（Andrew K.在两个DRS参加的工作组中，有机会在包括肉瘤计划，转化研究论坛，发育治疗论和双月份的实验室会议上进行互动和讨论。博士博士后培训计划和埃德娜·库基尔曼（Edna Cukierman）的主任墨菲（Murphy）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 解决目标1和2中提出的研究目标。在此期间，我将过渡到独立性，以寻求终身教师职位。在过去三年的资金中，我将继续在建立我的独立研究计划时在目标2和3中提出的研究。作为我未来的研究目标的一部分，我不仅对诸如IM等新型生物制剂的耐药机制感兴趣，而且还对建立模型和挑战现有范式的挑战以合理地开发组合疗法以减轻癌症的负担。 我已经在短暂的职业生涯中取得了重大的科学成就，并确立了自己成为GIST研究界的重要成员。我撰写了10篇研究文章和学术评论。我已经获得了许多奖项和培训奖学金，并在2009年的AACR和ASCO会议上被选为口头介绍我的作品，这是该提案的基础。