Melanoma Resistance to Apoptosis: Mechanisms and Therapeutic Potential

黑色素瘤对细胞凋亡的抵抗：机制和治疗潜力

• 批准号: 9031874
• 负责人: DAVID A. NORRIS
• 金额: --
• 依托单位: VA EASTERN COLORADO HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9031874
• 关键词: Alternative Therapies; Apoptosis; Apoptotic; BCL1 Oncogene; BCL2 gene; BRAF gene; Cell Count; Cell Death; Cell Line; Cells; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Death Domain; Disease; Drug Combinations; Effectiveness; FDA approved; Family; Family member; Fenretinide; Generations; Goals; Incidence; Knock-out; Lead; Libraries; Link; Malignant Neoplasms; Medicine; Melanoma Cell; Metastatic Melanoma; MicroRNAs; Military Personnel; Mission; Mitogen-Activated Protein Kinase Inhibitor; Modeling; Neoplasm Metastasis; Pathway interactions; Patient Care; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Population; Proteins; Relapse; Research; Resistance; Retinoids; Sampling; Services; Sun Exposure; Technology; Testing; Therapeutic; Time; United States; Xenograft Model; alternative treatment; base; cancer cell; gamma secretase; genome editing; implantation; in vitro testing; in vivo; inhibitor/antagonist; killings; melanoma; mutant; neoplastic cell; novel; novel therapeutic intervention; novel therapeutics; public health relevance; response; self-renewal; small molecule inhibitor; success; therapeutic miRNA; therapy resistant; treatment strategy; tumor

 DESCRIPTION (provided by applicant): Discovering new therapies for metastatic melanoma is still a pressing issue, given the lack of treatment options for wild type BRAF melanoma, frequent relapses and (with some drugs) limited responders, even with the recent exciting FDA approved drugs. The long-term goal of this project is to develop alternative treatments, which will lead to longer lasting clinical responses and better patient outcomes for patients with malignant melanomas. Specifically, this proposal aims to discover treatments that target anti-apoptotic defenses and pathways used by melanoma to resist apoptosis. Cancer initiating cells (CICs) are implicated in cancer cells' resistance to treatment, resulting in relapse. It is therefore crucial to identify treatment strateies that eliminate the bulk of melanoma cells (de-bulk) as well as the CIC populations of the melanoma (also known as Melanoma Initiating Cells, MICs). The BCL-2 family is important in regulating apoptosis. The interactions among the pro- and anti-apoptotic BCL-2 family members can control the initiation of intrinsic apoptosis. ABT-737 is a potent small molecule inhibitor of the anti-apoptotic proteins BCL-2/BCL-XL/BCL-W. We have identified two promising agents, the synthetic retinoid derivative, fenretinide (4-HPR) and a Gamma Secretase Inhibitor (GSI-I), that synergize with ABT-737 to inhibit multiple anti-apoptotic BCL-2 family members. When tested in vitro, in melanoma cell lines and tumor cells maintained in the patient derived xenograft (PDX) model, we found that ABT-737 combined with either drug partner not only de-bulked the melanoma cells, but also killed the CICs, for melanomas with either mutant or wild-type BRAF. Both combinations also dramatically limited tumor self-renewal. MiRNA-based therapies provide an exciting, novel therapeutic opportunity for many diseases. We show that replacement of miR-26a induced cell death in a subset of melanomas, through targeting the anti-apoptotic protein, silencer of death domain (SODD). Therefore, we propose to further demonstrate the efficacy of the two approaches of targeting anti-apoptotic defenses in melanoma and investigate the mechanisms involved: inhibiting multiple anti-apoptotic BCL-2 family members at once (Aims 1 and 2) and a miRNA-based approach to target SODD (Aim 3). Aim 1: Examine the mechanism(s) determining efficacy for the combinations of ABT-737 with 4-HPR or GSI in de-bulking melanoma and targeting MICs. We will define the mechanism of action for these drug combinations to eliminate MICs using CRISPR genomic editing technology, and further examine how the combinations induced MCl-1 degradation. {Aim 2: Investigate the efficacy and mechanisms of the combination treatments of ABT-737 with 4-HPR or GSI in vivo. We will determine the effects and examine the mechanisms involved by using a conventional xenograft models with knockout cell lines, a melanoma PDX model, a melanoma metastasis model, and a low-cell-number implantation xenograft model for assessing MIC-initiated tumor formation.} {Aim 3: Study the effects of miR-26a replacement in de-bulking melanoma and targeting MICs, and identify the candidate compounds as potential partners with miR-26a treatment. We will test the effects of miR-26a in targeting MICs as in Aim 1, and screen for potential partner compounds with an apoptosis compound library and a MAPK inhibitor library.} In summary, this proposal extends our exciting preliminary studies that have identified drug combinations which overcome melanoma resistance to apoptosis. We will further test these promising combinations with effects on de-bulking melanoma, and eliminating the MIC populations and limiting their self-renewal. We will also determine the effectiveness of an original miRNA-based therapy alone or in combination. Results are likely to identify new therapeutic approaches for treating melanoma, with unique effectiveness in killing MICs.

 描述（由申请人提供）： 鉴于野生型BRAF黑色素瘤缺乏治疗选择，经常继电器以及（某些药物）有限的反应者，即使是最近令人兴奋的FDA批准的药物，发现转移性黑色素瘤的新疗法仍然是一个紧迫的问题。该项目的长期目标是开发替代治疗，这将导致更长的临床反应和恶性黑色素瘤患者的患者结局更好。具体而言，该提案旨在发现靶向抗凋亡防御的治疗方法和黑色素瘤用于抵抗细胞凋亡的途径。癌细胞对治疗的抵抗力中暗示了癌症启动细胞（CIC），从而缓解。因此，至关重要的是确定消除大部分黑色素瘤细胞（De-bulk）以及黑色素瘤的CIC种群（也称为黑色素瘤引发细胞，MICS）至关重要。 Bcl-2家族在调节凋亡方面很重要。促凋亡和抗凋亡的Bcl-2家族成员之间的相互作用可以控制内在凋亡的开始。 ABT-737是抗凋亡蛋白Bcl-2/bcl-XL/bcl-W的潜在小分子抑制剂。我们已经确定了两种有前途的药物，即合成性维生素衍生物，富烯蛋白（4-HPR）和一个伽马泌尿酶抑制剂（GSI-I），它们与ABT-737协同抑制了多个抗凋亡BCL-2家族成员。当在体外测试时，在患者衍生的特征（PDX）模型中维持的黑色素瘤细胞系和肿瘤细胞中，我们发现，ABT-737与吸毒伴侣相结合，不仅脱离了黑色素瘤细胞，而且还杀死了CICS，还杀死了具有突变体或野生型Braf的黑色素瘤。两种组合也极有限地限制了肿瘤自我更新。基于miRNA的疗法为许多疾病提供了令人兴奋的新疗法机会。我们表明，通过靶向抗凋亡蛋白，死亡结构域的消音器（SODD），miR-26a的替换在黑色素瘤的一部分中诱导细胞死亡。因此，我们建议进一步证明靶向黑色素瘤中抗凋亡防御的两种方法的效率，并研究所涉及的机制：立即抑制多个抗凋亡Bcl-2家族成员（AIMS 1和2）和一种基于miRNA的靶向SODD方法（AIM 3）。 AIM 1：检查ABT-737与4-HPR或GSI组合的有效性的机制，在脱毛的黑色素瘤和靶向MIC中的有效性。我们将定义这些药物组合使用CRISPR基因组编辑技术消除MIC的作用机理，并进一步研究组合如何诱导MCL-1降解。 {AIM 2：研究ABT-737与4-HPR或GSI的组合处理的效率和机制。我们将确定效果，并通过使用与基因敲除细胞系，黑色素瘤模型，黑色素瘤转移模型以及低细胞数量的植入型Xen摄影模型来评估MIC启动的肿瘤形成的传统型型模型，用于评估MIC-26A替补群的影响，并定位了针对Mir-26A替代品的甲壳虫，并定位了针对MIC的效果。使用miR-26a处理。我们将测试miR-26a在靶向MIC中的影响，如AIM 1，并筛选出具有凋亡化合物库和MAPK抑制剂库的潜在伴侣化合物。}总而言之，该提案扩展了我们令人兴奋的初步研究，这些研究已经确定了克服了对凋亡的黑色素瘤抗性的药物组合。我们将进一步测试这些承诺的组合与对脱毛黑色素瘤的影响，并消除MIC种群并限制其自我更新。我们还将单独或组合使用原始的基于miRNA的治疗的有效性。结果可能会确定用于治疗黑色素瘤的新治疗方法，并在杀死MIC方面具有独特的有效性。