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.

项目概要/摘要 甲磺酸伊马替尼 (IM) 彻底改变了胃肠道间质瘤患者的治疗 （胃肠道间质瘤）；然而，尽管初步观察到了疗效，但 IM 的临床耐药性已成为现实。 此外，对于治疗开始时难治的患者（~20%）来说，选择非常有限。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本，我确定了 32 基因特征，包括可预测 IM 反应的 KRAB-ZNF 91 亚家族成员。我 随后证明，其中许多基因不仅可以预测 IM 反应，还可以介导 药物的活性。为了从机制上确定这些 ZNF 如何调节对 IM 的反应， RNAi 方法用于敲低 GIST 中预测特征（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谱分析。这导致发现骨膜蛋白， NEDD9 和 TGFβ3 在 siRNA 介导的敲低后普遍下调。这很有趣 因为 TGFβ3 是骨膜素和 NEDD9 的已知诱导剂，这两种物质均已被频繁报道 在多种人类癌症中过度表达，表明这些临床研究中发现的 ZNF 研究可能是这些途径的“主控制器”。有趣的是，缺氧已被证明 上调 NEDD9 和骨膜素的表达（通过 PI3K），导致攻击性增加 并增强肿瘤细胞的存活率。因此，periostin 和 NEDD9 可能都在 TGFβ 和缺氧的类似机制，由 ZNF 控制，调节 GIST 细胞的 IM 反应。 NEDD9 也被证明在 IM 抗性 GIST 细胞系中过度表达且 SRC 过度激活， NEDD9 的敲除可恢复 IM 敏感性。我推测 GIST 中的 IM 耐药可能与 缺氧导致 ZNF 反式激活 TGFβ3，从而导致随后诱导骨膜素和/或 NEDD9 和 PI3-K 或 SRC 通路的激活分别。激活这些潜在的“救援” “路径”的发生独立于 KIT/PDGFRA 信号传导，从而克服了 IM 对亲- 生存途径。我的初步数据表明这些 ZNF 负责调节这条通路 这些基因的过度表达与 GIST 患者对 IM 治疗的临床耐药性相关。 为了检验我的假设，我在目标 1 中建议确定 TGFβ3、NEDD9 和/或骨膜素的表达是否 原发性 GIST 样本中的 ZNF 与 IM 反应（短期和长期）和 ZNF 表达相关。目标 2 I 将研究这些 ZNF 如何影响 TGFβ3、NEDD9 和骨膜素表达以及最终对 IM 的反应。 目标 3 将确定是否针对缺氧诱导的 TGFβ3→骨膜素→PI-3K/AKT 和/或缺氧- 体内诱导的TGFβ3→NEDD9→SRC拯救途径将消除对IM的抵抗。的中心宗旨 该建议是，当同时使用靶向药物组合使用时，靶向药物将变得有效 多种致癌途径，防止规避可充当“救援路线”的平行途径 对于这些肿瘤细胞。 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 (FCCC) 完成。 FCCC是国家癌症研究所指定的综合性癌症中心，全国认可 表彰其在医学、放射和肿瘤外科领域的领导地位。 FCCC 领导层致力于提供 必要的基础设施和支持，让我成功完成我拟议的研究。我将受到指导 由博士。 Margaret von Mehren（肉瘤项目教授兼主任）、Andrew K. Godwin（教授和肉瘤项目主任） 堪萨斯大学医学院 (KUMC) 分子肿瘤学主任 Erica Golemis （FCCC 教授兼副首席科学官）、FCCC 和 KUMC 尊敬的教职人员以及 国际公认的转化研究和肿瘤医学领域的专家。我将会有 有机会在包括肉瘤在内的工作组定期会议上互动和讨论我的研究 参加项目、转化研究论坛、发展治疗学和每两个月一次的实验室会议 由两位博士。冯·梅伦和戈德温（通过视频会议）。我的首要目标是获得独立的 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 教员组成（博士后主任 Maureen Murphy 博士） 培训计划和 Edna Cukierman），以便审查我的研究进展并提供反馈。 在资助的前两年，我将继续与我的导师密切合作来解决研究问题 目标 1 和 2 中提出的目标。在此期间，我将过渡到独立，寻求终身职位 教职职务。在过去三年的资助中，我将继续目标 2 和 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 仅限于对新型生物制剂（例如IM）的耐药机制，但在建立模型和挑战 合理开发联合疗法以减轻癌症负担的现有范式。 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 我自己成为 GIST 研究界的重要成员。我撰写了 10 篇研究文章 学术评论。我获得了多项奖项和培训奖学金，并被选为口头讲师 在 2009 年 AACR 和 ASCO 会议上介绍我的工作，即该提案的基础。