DIFFERENTIATION AND APOPTOSIS IN MALIGNANT NEUROBLASTOMA

恶性神经母细胞瘤的分化和凋亡

• 批准号: 7271142
• 负责人: SWAPAN K. RAY
• 金额: $ 35.44万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-02 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7271142
• 关键词: 1 year old; Acids; Adrenal Glands; Aftercare; Animal Model; Apigenin; Apoptosis; Apoptotic; Biochemical; Calpain; Caspase; Cell Differentiation process; Cell Line; Cells; Cessation of life; Characteristics; Child; Childhood; Combined Modality Therapy; Cysteine Protease; DNA Fragmentation; Diagnosis; Differentiation Therapy; Disease; Dose; Down-Regulation; Epigallocatechin Gallate; Evaluation; Family; Family member; Fenretinide; Flavonoids; Gene Expression; Genistein; Growth; Human; Induction of Apoptosis; Infant; Inhibition of Apoptosis; Investigation; Isotretinoin; Malignant - descriptor; Malignant Childhood Neoplasm; Messenger RNA; Molecular; Mus; Neuroblastoma; Neuronal Differentiation; Neurons; Nuclear; Nude Mice; Outcome; Pathway interactions; Patients; Physical condensation; Process; Protein Family; Proteins; Proteolysis; Research Personnel; Retinoids; Staining method; Stains; Sympathetic Nervous System; Telomerase; Testing; Therapeutic; Time; Treatment Protocols; Tretinoin; Up-Regulation; Xenograft procedure; base; early childhood; efficacy evaluation; egg; gallocatechol; inhibitor-of-apoptosis protein; innovation; older patient; outcome forecast; programs; response; retinamide; success; tumor

DESCRIPTION (provided by applicant): Neuroblastoma is the most common malignant disease of early childhood. Infants younger than 1 year have a good prognosis, while older patients show metastatic neuroblastoma with poor outcome and death. Because current treatment strategies are ineffective in children older than 1 year, innovative therapeutic strategies are urgently needed. This proposal aims to develop a combination therapy to control the growth of malignant neuroblastoma. Our preliminary results indicate that retinoids, such as all-trans-retinoic acid (ATRA), 13-cis-retinoicacid (13-CRA), and fenretinide [N-(4-hydroxyphenyl) retinamide or 4-HPR] induce differentiation in 5 human neuroblastoma cell lines (SH-SY5Y, SK-N-BE2, SK-N-DZ, IMR-32, and SK-N-MC). Also, flavonoids, such as apigenin (APG), (-)-epigallocatechin (EGG), (-)-epigallocatechin-3-gallate (EGCG), and genistein (GST), induce apoptosis. Interestingly, neuroblastoma cells differentiated with specifically 4- HPR became more sensitive to flavonoids, especially GST, for induction of apoptosis with increased ratio of bax:bcl-2 and upregulation of calpain and caspases. Treatment of neuroblastoma xenografts in nude mice with a combination of low doses of 4-HPR and GST regressed tumors with induction of differentiation and apoptosis. We hypothesize that (1) 4-HPR induces differentiation and down regulates survival factors; (2) GST induces apoptosis inhibiting survival factors and activating cysteine proteases and enhances apoptosis in differentiated cells; (3) treatment of neuroblastoma xenografts with 4-HPR and GST induces differentiation and enhances apoptosis. Our central hypothesis is that promotion of differentiation with a selective retinoid down regulates telomerase and survival proteins, and sensitizes differentiated cells to a selective flavonoid for enhancing apoptosis with activation of cysteine proteases. The objective is to use a combination therapy for differentiation and apoptosis in neuroblastoma cells in culture as well in ectopic and orthotopic xenografted animal models. To achieve this objective, we propose the following specific aims: Specific Aim 1: To determine the morphological and biochemical features of neuronal differentiation in five human neuroblastoma cell lines in response to selected retinoids. Specific Aim 2: To examine the induction of apoptosis in five human neuroblastoma cell lines following treatment with selected flavonoids and enhancement of apoptosis in differentiated cells. Specific Aim 3: To determine the efficacy of combination of a selected retinoid and a selected flavonoid for induction of differentiation and enhancement of apoptosis in neuroblastoma in ectopic and orthotopic xenografts in athymic nude mice. Success of our therapeutic strategy can form a basis for treatment of neuroblastoma in humans.

描述（由申请人提供）：神经母细胞瘤是幼儿期最常见的恶性疾病。 1 岁以下的婴儿预后良好，而年龄较大的患者则表现为转移性神经母细胞瘤，预后较差甚至死亡。由于目前的治疗策略对1岁以上儿童无效，因此迫切需要创新的治疗策略。该提案旨在开发一种联合疗法来控制恶性神经母细胞瘤的生长。我们的初步结果表明，类视黄醇，如全反式视黄酸 (ATRA)、13-顺式视黄酸 (13-CRA) 和芬维A胺 [N-(4-羟基苯基) 视黄酰胺或 4-HPR] 可诱导 5 种细胞的分化。人神经母细胞瘤细胞系（SH-SY5Y、SK-N-BE2、SK-N-DZ、IMR-32 和 SK-N-MC）。此外，黄酮类化合物，如芹菜素 (APG)、(-)-表没食子儿茶素 (EGG)、(-)-表没食子儿茶素-3-没食子酸酯 (EGCG) 和染料木黄酮 (GST)，可诱导细胞凋亡。有趣的是，用 4-HPR 特异性分化的神经母细胞瘤细胞对黄酮类化合物（尤其是 GST）变得更加敏感，通过增加 bax:bcl-2 比例以及上调钙蛋白酶和半胱天冬酶来诱导细胞凋亡。用低剂量的 4-HPR 和 GST 联合治疗裸鼠神经母细胞瘤异种移植物，可消退肿瘤并诱导分化和凋亡。我们假设（1）4-HPR诱导分化并下调生存因子； (2)GST诱导细胞凋亡，抑制生存因子并激活半胱氨酸蛋白酶，增强分化细胞的凋亡； (3)用4-HPR和GST处理神经母细胞瘤异种移植物诱导分化并增强细胞凋亡。我们的中心假设是，用选择性类视黄醇促进分化下调端粒酶和存活蛋白，并使分化细胞对选择性类黄酮敏感，从而通过激活半胱氨酸蛋白酶来增强细胞凋亡。目的是使用联合疗法来培养神经母细胞瘤细胞以及异位和原位异种移植动物模型中的分化和凋亡。为了实现这一目标，我们提出以下具体目标： 具体目标 1：确定五种人神经母细胞瘤细胞系对选定的类维生素A反应的神经元分化的形态和生化特征。具体目标 2：检查用选定的类黄酮处理后五种人神经母细胞瘤细胞系中细胞凋亡的诱导以及分化细胞中细胞凋亡的增强。具体目标3：确定选定的类维生素A和选定的类黄酮的组合对无胸腺裸鼠异位和原位异种移植物中神经母细胞瘤诱导分化和增强细胞凋亡的功效。我们的治疗策略的成功可以为人类神经母细胞瘤的治疗奠定基础。