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(申请人提供)：甲磺酸伊马替尼(IM)使胃肠道间质瘤(GIST)患者的治疗发生了革命性变化；然而，尽管观察到初步疗效，临床对IM的抵抗已经成为现实。此外，对于在治疗开始时难治的患者(~20%)来说，选择非常有限。使用临床预 在一项预期的新辅助II期试验(RTOG 0132)的治疗活检样本中，我发现了一个32基因签名，其中包括可以预测IM应答的KRAB-ZNF91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的反应，还可以调节药物的活性。为了从机制上确定这些ZNF可能如何调节IM的反应，使用RNAi方法敲除GIST细胞中预测信号(包括10个ZNF)内的基因，并使用外显子1.0 ST阵列进行表达谱分析。这导致了研究发现，在siRNA介导的基因敲除后，Periostin、NEDD9和TGF23普遍下调。这很有趣，因为TGF23是Periostin和NEDD9的已知诱导者，据报道，这两种基因在各种人类癌症中经常过度表达，并表明从这些临床研究中发现的这些ZNF可能是这些通路的“主控制器”。有趣的是，低氧已被证明上调NEDD9和Periostin的表达(通过PI3K)，从而增加肿瘤细胞的侵袭性和提高其存活率。因此，在GIST细胞中，PERIOSIN和NEDD9可能以相似的机制受TGF2和ZNFS控制的低氧调节IM反应。在IM耐药的GIST细胞系中，NEDD9也高表达，SRC高激活，NEDD9的敲除恢复了IM的敏感性。我推测，GIST的IM抵抗可能与缺氧导致ZNF反式激活TGF23，导致随后分别诱导Periostin和/或NEDD9以及激活PI3-K或SRC通路有关。这些潜在的“救援途径”的激活不依赖于KIT/PDGFRA信号，从而克服了IM在促生存途径上的抑制活性。我的初步数据表明，这些ZNF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中建议确定GIST原发样本中TGF23、NEDD9和/或Periostin的表达是否与IM反应(短期和长期)和ZNF表达相关。在目标2中，我将研究这些ZNF如何影响TGF23、NEDD9和Periostin的表达以及最终对IM的反应。目的3确定体内靶向低氧诱导的TGF23-&GT；Periostin-&GT；PI-3K/AKT和/或低氧诱导的TGF23-&GT；NEDD9-&GT；SRC救援通路是否能消除对IM的耐药性。这项提议的中心原则是，当靶向药物联合使用时，将变得有效，同时阻断多个致癌途径，防止绕过可作为这些肿瘤细胞的“拯救途径”的平行途径。 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，因其在内科、放射和外科肿瘤学方面的领先地位而在全国范围内得到认可。Fccc领导层致力于为我成功完成拟议的学习提供必要的基础设施和支持。我的导师包括玛格丽特·冯·迈伦博士(肉瘤项目教授兼主任)、安德鲁·K·戈德温博士(堪萨斯大学医学院分子肿瘤学教授兼主任)和埃丽卡·戈莱米斯博士(中国社会科学院副教授兼首席科学官)，他们都是世界知名的翻译研究和医学肿瘤学专家。我将有机会在工作组的定期会议上互动和讨论我的研究，这些工作组包括肉瘤计划、转化研究论坛、发展治疗以及冯·梅伦博士和戈德温博士参加的每两个月一次的实验室会议(通过视频会议)。我的主要目标是在癌症研究方面获得独立的学术生涯。为了帮助实现这一目标，我成立了一个顾问委员会，成员包括我的导师和fcc的其他教职员工(博士后培训项目主任Maureen Murphy博士和Edna Cukierman博士)，以审查我的研究进展并提供反馈。在资助的头两年，我将继续与我的导师密切合作，以 解决目标1和目标2中提出的研究目标。在这段时间里，我将过渡到独立，寻找一个终身教职的职位。在最后三年的资助中，我将继续目标2和目标3中提出的学习，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，我不仅对IM等新生物药物的耐药机制感兴趣，而且对建立模型和挑战现有范式以合理开发联合疗法以减轻癌症负担感兴趣。 在我短暂的职业生涯中，我已经取得了重大的科学成就，并成为GIST研究界的重要成员。我撰写了10篇研究文章和学术评论。我获得了许多奖项和培训奖学金，并被选中在2009年的AACR和ASCO会议上口头介绍我的工作，这是这项提议的基础。