Targeting EWS/FLI1-driven pathways to improve therapeutic gains in Ewing's Sarcom

靶向 EWS/FLI1 驱动的途径以提高尤文肉瘤的治疗效果

• 批准号: 7649300
• 负责人: VICENTE NOTARIO
• 金额: $ 31.85万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7649300
• 关键词: Adolescent; Affect; Aftercare; Antineoplastic Agents; Arts; Askin' s tumor; Bone neoplasms; Cadherins; Cell Line; Cells; Child; Chromosomes, Human, Pair 11; Complement component C1s; Complex; Data; Disease; Disease Progression; Dose; Down-Regulation; E-Cadherin; Equilibrium; Ewings sarcoma; FUS-1 Protein; Family; Genes; Health; High Dose Chemotherapy; Human Chromosomes; In Vitro; Individual; Ionizing radiation; Laboratories; Late Effects; Localized Disease; Malignant - descriptor; Malignant Bone Neoplasm; Malignant Childhood Neoplasm; Malignant Neoplasms; Mediating; Modality; Modeling; Molecular; Mus; Neoplasm Metastasis; Normal tissue morphology; Operative Surgical Procedures; PARP inhibition; Pathway interactions; Patients; Pharmaceutical Preparations; Phospholipase D; Phosphotransferases; Play; Poly(ADP-ribose) Polymerases; Primary Neoplasm; Primitive Neuroectodermal Tumor; Production; Property; Protein Kinase; Protein Kinase C Alpha; Proteins; Protocols documentation; Public Health; Radiation; Radiation therapy; Radiosensitization; Relapse; Reporting; Resistance; Role; Signal Pathway; Signal Transduction; Snails; Solid; Specificity; Techniques; Therapeutic; Tumorigenicity; Xenograft procedure; bone; caveolin 1; chemotherapy; cohort; design; follow-up; genetic regulatory protein; human FRAP1 protein; hybrid gene; improved; inhibitor/antagonist; killings; mTOR Inhibitor; member; neoplastic; neoplastic cell; novel; outcome forecast; phospholipase D2; prototype; radiation resistance; reconstitution; research study; response; soft tissue; tool; transcription factor; treatment strategy; tumor; tumor growth; young adult

DESCRIPTION (provided by applicant): Tumors of the Ewing's sarcoma family (ESFT) are solid, highly malignant neoplasms of the bone and soft tissues that most often affect children and adolescents, being the second most common bone malignancies among young adults. Current treatment includes a combined modality with chemotherapy and radiotherapy. However, there are two important problems to consider: a) although ESFT are generally responsive to treatment, the overall cure rate is low because these tumors are very aggressive and patients frequently present with metastatic disease; and b) exposure of young patients to high doses of chemotherapy and/or radiation is frequently associated with a variety of adverse health effects that in some cases do not develop until many years after treatment completion. Consequently, treatment strategies are needed to maximize curability while simultaneously minimizing adverse late effects for the patients. In preliminary studies, we directly targeted EWS/FLI-1, the transcription factor known to be responsible for the malignant properties of most ESFT, using molecular tools designed to block its activity, either alone or in combination with chemotherapeutic drugs and/or radiation. Although experiments in vitro and in mouse xenografts demonstrated that these tools efficiently delayed tumor growth, it became clear that simultaneous targeting of additional molecules that act downstream of EWS/FLI-1 would be necessary to maximize antitumor activity and to allow the use of lower therapeutic doses, thus minimizing negative late effects as much as possible. In this context, the main objective of this proposal is to exploit a novel EWS/FLI-1-driven pathway recently established in our laboratory (EWS/FLI-1-[Caveolin->1-Snail-E-cadherin]) to sensitize ESFT cells to therapy, using ionizing radiation (IR) as a model anti-neoplastic agent that does not suffer from the specificity- related problems that frequently complicate studies with chemotherapeutic drugs. Since the extent of IR related late effects in normal tissues surrounding the tumor are directly related to the IR dose, in pediatric cancers it is important to employ the lowest dose possible to cure the tumors. Using EWS cells as the ESFT prototype, our central hypothesis is that (a) targeting caveolin-1 (CAV1) itself and/or its interactions with signaling or regulatory proteins relevant for ESFT molecular pathobiology (such as phospholipase D2 and protein kinase C1) will render EWS cells more sensitive to IR, and (b) that this response may be further improved by simultaneously targeting components of other pathways also known to radiosensitize EWS cells, such as poly (ADP-ribose) polymerase (PARP). Because we already identified CAV1 as a direct transcriptional target of EWS/FLI-1 and a key determinant of the tumorigenicity and chemotherapeutic response of EWS cells, targeting CAV1 signaling may have a dual effect: enhancing killing of EWS cells by low-dose IR and down- regulating their neoplastic properties. PUBLIC HEALTH RELAVANCE: Tumors of the Ewing's sarcoma family (ESFT) are solid, highly malignant cancers of the bone and soft tissues that most often affect children and adolescents, being the second most common bone malignancies among young adults. There are two important problems to consider: a) despite aggressive treatment strategies, such as high-dose chemotherapy combined with surgery and/or extended radiotherapy, the prognosis for ESFT patients with large primary tumors or metastatic disease remains poor; about 50% of patients eventually relapse, even after five years, and more than 30% of patients with localized disease and about 80% of patients with metastases die due to disease progression; and b) exposure of young patients to high doses of chemotherapy and/or radiation is frequently associated with a variety of adverse health effects that in some cases do not develop until many years after treatment completion. Consequently, treatment strategies are needed to maximize curability while simultaneously minimizing adverse late effects for the patients. In this context, we propose to use Ewing's sarcoma cells as the ESFT prototype, and radiation as a model anti-cancer agent that does not suffer from the specificity-related problems that frequently complicate studies with chemotherapeutic drugs, to devise protocols to sensitize ESFT cells to therapy. Specifically, we will use state-of-the-art techniques to block the production and/or activity of molecular components of a novel pathway {EWS/FLI-1-> [CAV1-PKC1-PLD2]-mTOR} established in our laboratory as driven by the EWS/FLI-1 protein, which is known to be responsible for the malignant properties of ESFT. Our preliminary studies strongly indicate that components of this pathway such as CAV1 and PKC1 contribute to render ESFT cells resistant to chemotherapy and radiation. Our approach will, first, characterize the mechanism of action of each of the pathway components and their interrelationships, and then target them either individually or in combination to improve the therapeutic response of the ESFT cells and tumors. Treatments with inhibitors of PARP-1, a molecule previously shown in our Department to contribute to the radiation resistance of ESFT, will be included also in our experimental protocols.

描述（由申请方提供）：尤文氏肉瘤家族肿瘤（ESFT）是骨和软组织的实体、高度恶性肿瘤，最常累及儿童和青少年，是年轻成人中第二常见的骨恶性肿瘤。目前的治疗包括化疗和放疗的联合治疗。然而，有两个重要的问题需要考虑：a）尽管ESFT通常对治疗有反应，但总体治愈率较低，因为这些肿瘤非常具有侵袭性，并且患者经常出现转移性疾病;和B）年轻患者暴露于高剂量化疗和/或或辐射通常与各种不良健康影响有关，在某些情况下，这些不良健康影响直到治疗完成多年后才出现。因此，需要治疗策略，以最大限度地提高治愈率，同时最大限度地减少对患者的不良晚期影响。在初步研究中，我们直接靶向EWS/FLI-1，这是已知负责大多数ESFT恶性特性的转录因子，使用设计用于阻断其活性的分子工具，单独或与化疗药物和/或放射联合使用。尽管体外和小鼠异种移植物中的实验证明这些工具有效地延迟了肿瘤生长，但很明显，同时靶向作用于EWS/FLI-1下游的其他分子对于最大化抗肿瘤活性和允许使用较低的治疗剂量是必要的，从而尽可能地减少负面的晚期效应。在这种情况下，该提议的主要目的是利用我们实验室最近建立的新的EWS/FLI-1驱动的途径（EWS/FLI-1-[小窝蛋白->1-蜗牛-E-钙粘蛋白]）来使ESFT细胞对治疗敏感，使用电离辐射（IR）作为模型抗肿瘤剂，其不遭受特异性相关的问题，所述特异性相关的问题经常使化学治疗药物的研究复杂化。由于肿瘤周围正常组织中IR相关迟发效应的程度与IR剂量直接相关，因此在儿科癌症中，重要的是采用可能治愈肿瘤的最低剂量。使用EWS细胞作为ESFT原型，我们的中心假设是（a）靶向小窝蛋白-1（CAV 1）本身和/或其与ESFT分子病理学相关的信号或调节蛋白的相互作用，（如磷脂酶D2和蛋白激酶C1）将使EWS细胞对IR更敏感，和（B）通过同时靶向也已知使EWS细胞放射增敏的其它途径的组分，如聚（ADP-核糖）聚合酶（PARP），可以进一步改善这种应答。因为我们已经将CAV 1鉴定为EWS/FLI-1的直接转录靶标和EWS细胞的致瘤性和化疗反应的关键决定因素，所以靶向CAV 1信号传导可能具有双重作用：通过低剂量IR增强对EWS细胞的杀伤并下调其肿瘤性质。公共卫生关系：尤文氏肉瘤家族肿瘤（ESFT）是骨和软组织的实体、高度恶性癌症，最常影响儿童和青少年，是年轻人中第二常见的骨恶性肿瘤。有两个重要的问题需要考虑：a）尽管有积极的治疗策略，如高剂量化疗联合手术和/或延长放疗，但患有大原发性肿瘤或转移性疾病的ESFT患者的预后仍然很差;大约50%的患者最终会复发，甚至在五年后，超过30%的局限性疾病患者和约80%的转移性疾病患者因疾病进展而死亡;和B）年轻患者暴露于高剂量化疗和/或或辐射通常与各种不良健康影响有关，在某些情况下，这些不良健康影响直到治疗完成多年后才出现。因此，需要治疗策略，以最大限度地提高治愈率，同时最大限度地减少对患者的不良晚期影响。在这种情况下，我们建议使用尤因肉瘤细胞作为ESFT原型，辐射作为一种模型抗癌剂，不遭受的特异性相关的问题，经常复杂化的研究与化疗药物，设计方案，使ESFT细胞敏感的治疗。具体而言，我们将使用最先进的技术来阻断我们实验室中建立的由EWS/FLI-1蛋白驱动的新途径{EWS/FLI-1-> [CAV 1-PKC 1-PLD 2]-mTOR}的分子组分的产生和/或活性，已知EWS/FLI-1蛋白是ESFT恶性特性的原因。我们的初步研究强烈表明，该途径的组分，如CAV 1和PKC 1有助于使ESFT细胞对化疗和放疗产生抗性。我们的方法将首先表征每个途径组分的作用机制及其相互关系，然后单独或组合靶向它们，以改善ESFT细胞和肿瘤的治疗反应。我们的实验方案中也将包括PARP-1抑制剂的治疗，PARP-1是我们部门先前显示的有助于ESFT辐射抗性的分子。