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（由申请人提供）：尤文氏肉瘤家族肿瘤（ESFT）是骨和软组织的高度恶性的实性肿瘤，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。目前的治疗包括化疗和放疗的联合治疗。然而，有两个重要问题需要考虑：a）虽然 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； b) 年轻患者接受高剂量化疗和/或放疗通常会带来各种不良健康影响，在某些情况下，这些影响直到治疗完成多年后才会出现。因此，需要采取治疗策略来最大限度地提高治愈率，同时最大限度地减少对患者的后期不良影响。在初步研究中，我们直接针对 EWS/FLI-1（已知导致大多数 ESFT 恶性特性的转录因子），使用旨在阻断其活性的分子工具，无论是单独使用还是与化疗药物和/或放射组合使用。尽管体外和小鼠异种移植实验证明这些工具有效地延迟了肿瘤生长，但很明显，为了最大限度地提高抗肿瘤活性并允许使用较低的治疗剂量，有必要同时靶向作用于 EWS/FLI-1 下游的其他分子。剂量，从而尽可能减少负面的后期影响。在这种情况下，本提案的主要目标是利用我们实验室最近建立的一种新的 EWS/FLI-1 驱动途径 (EWS/FLI-1-[Caveolin->1-Snail-E-cadherin]) 来敏化ESFT 细胞使用电离辐射 (IR) 作为模型抗肿瘤药物进行治疗，该药物不会遇到通常使化疗药物研究复杂化的特异性相关问题。由于肿瘤周围正常组织中与 IR 相关的后期影响的程度与 IR 剂量直接相关，因此在儿科癌症中，采用尽可能低的剂量来治愈肿瘤非常重要。使用 EWS 细胞作为 ESFT 原型，我们的中心假设是，(a) 靶向 Caveolin-1 (CAV1) 本身和/或其与 ESFT 分子病理学相关信号或调节蛋白（例如磷脂酶 D2 和蛋白激酶 C1）的相互作用将使 EWS 细胞对 IR 更加敏感，并且 (b) 通过同时靶向也已知使 EWS 细胞放射增敏的其他途径的成分，例如聚 (ADP-核糖) 聚合酶，可以进一步改善这种反应（PARP）。因为我们已经确定 CAV1 是 EWS/FLI-1 的直接转录靶标，也是 EWS 细胞致瘤性和化疗反应的关键决定因素，因此靶向 CAV1 信号传导可能具有双重作用：通过低剂量 IR 增强对 EWS 细胞的杀伤作用，下调其肿瘤特性。公众健康相关性：尤文氏肉瘤家族 (ESFT) 肿瘤是骨和软组织的高度恶性的实体癌症，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。有两个重要问题需要考虑：a）尽管采取了积极的治疗策略，例如大剂量化疗联合手术和/或延长放疗，但患有大原发肿瘤或转移性疾病的 ESFT 患者的预后仍然很差；约50%的患者最终会复发，甚至在五年后，超过30%的局部疾病患者和约80%的转移患者因疾病进展而死亡； b) 年轻患者接受高剂量化疗和/或放疗通常会带来各种不良健康影响，在某些情况下，这些影响直到治疗完成多年后才会出现。因此，需要采取治疗策略来最大限度地提高治愈率，同时最大限度地减少对患者的后期不良影响。在这种情况下，我们建议使用尤文氏肉瘤细胞作为 ESFT 原型，并使用放射作为模型抗癌药物，以设计使 ESFT 细胞敏感的方案，该药物不会遇到经常使化疗药物研究复杂化的特异性相关问题。来治疗。具体来说，我们将使用最先进的技术来阻断我们实验室建立的新途径{EWS/FLI-1-> [CAV1-PKC1-PLD2]-mTOR}分子成分的产生和/或活性由 EWS/FLI-1 蛋白驱动，已知该蛋白是 ESFT 恶性特性的原因。我们的初步研究强烈表明，该途径的成分（例如 CAV1 和 PKC1）有助于使 ESFT 细胞对化疗和放疗产生抵抗。我们的方法将首先描述每个途径成分的作用机制及其相互关系，然后单独或组合地针对它们以改善 ESFT 细胞和肿瘤的治疗反应。 PARP-1 抑制剂的治疗也将包括在我们的实验方案中，PARP-1 是我们部门先前证明有助于 ESFT 抗辐射的分子。