SOMATIC NON-SYNONYMOUS MUTATIONS AS UNIQUE TUMOR ANTIGENS IN MELANOMA

体细胞非同义突变作为黑色素瘤中独特的肿瘤抗原

• 批准号: 8585662
• 负责人: Beatriz M. Carreno
• 金额: $ 19.84万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8585662
• 关键词: Address; Affect; Algorithms; Antigen Targeting; Antigens; Area; Binding; Biological Assay; Clinical; Clinical Trials; Computer Simulation; DNA; Data; Dendritic Cells; Development; Disease; Drug resistance; Gene Expression Profile; Genomics; HLA A*0201 antigen; Hybrids; Immune; Immunity; Immunotherapy; In Vitro; Incidence; Institutional Review Boards; Interferons; Interleukin-12; Intervention; Lead; Life; MAP Kinase Gene; Malignant Neoplasms; Medical; Metastatic Melanoma; Methods; Missense Mutation; Mutation; Nature; Neoplastic Cell Transformation; Nucleic Acids; Outcome; Patients; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phase; Phase I Clinical Trials; Pre-Clinical Model; Proteins; Protocols documentation; RNA; RNA Caps; Resistance; Signal Transduction; Somatic Mutation; T cell therapy; T-Lymphocyte; TNFSF5 gene; Testing; Therapeutic; Tumor Antigens; Ultraviolet Rays; Universities; Vaccination; Vaccines; Washington; base; cancer genome; cancer immunotherapy; combinatorial; exome; gp100 Antigen; improved; inhibitor/antagonist; insight; melanocyte; melanoma; next generation; novel strategies; peptide I; public health relevance; research study; response; synthetic peptide; transcriptome sequencing; tumor; tumor progression

DESCRIPTION (provided by applicant): The incidence of melanoma continues to rise worldwide with an estimated 76,250 new cases in the US for 2012. Despite the advent of new therapies, melanoma remains an incurable malignancy and thus, represents a disease area of unmet medical need. Melanoma is notable for its association with early-in-life UV-light exposure, highest mutational rate (14-30 per Mb) among cancer genomes and ability to induce spontaneous T cell immunity. The most common (60%) somatic non-synonymous mutation in melanoma is BRAFV600E. Targeted therapies, such as vemurafenib, directed at inhibiting this driver mutation have resulted in high responses rates, albeit of limited duration due to acquisition of drug resistance. In contrast, immune-based therapies have yielded low response rates that are often durable. Emerging data suggest that BRAFV600E inhibition, through paradoxical increased MAPK signaling, potentiates anti-tumor T cell immunity. Altogether, these findings support testing vemurafenib in combination with immunotherapies to improved clinical outcomes. Among immunotherapies, investigational vaccines and adoptive T cell therapies (ACT) are beginning to show efficacy in early phase clinical trials. Our recent pilot phase 1 clinical trial using CD40L/IFN-? matured dendritic cells (DC) and melanocyte lineage-restricted gp100 antigen have revealed the therapeutic benefit of IL-12, produced by DC, in vaccination of patients with metastatic melanoma. However, a critical barrier to development of improved vaccines and ACT is the nature and paucity of validated melanoma antigens. To date, antigens targeted for immune intervention are predominantly derived from structurally unaltered germline/ lineage/differentiation proteins and have demonstrated limited clinical benefit. Novel strategies are needed to identify patient-specific/unique tumor antigens in order to develop the next generation of personalized cancer immunotherapies. Experimental evidence in pre-clinical models supports the thesis that these unique tumor antigens, primarily arising during neoplastic transformation, can elicit T cell immunity capable of protecting the host from cancer progression. The emergence of hybrid capture transcriptome sequencing (RNA-capture sequencing, RNA-cap seq) offers a sensitive method to interrogate cancer genomes for expressed somatic mutations and assess their level of expression. Our ongoing experiments studying the melanoma transcriptome, have found over 500 expressed somatic non-synonymous mutations per tumor. We propose to use RNA-cap seq along with in-silico HLA class I peptide binding prediction algorithms and in vitro T cell assays to test the hypothesis that somatic non- synonymous mutations give rise to unique melanoma antigens and that acquisition of resistance to BRAFV600E inhibition is accompanied by changes in the melanoma antigenic landscape. This hypothesis will be addressed in the experiments of the following Specific Aims: (1) Characterize the repertoire of melanoma expressed somatic mutations and evaluate their potential as unique antigens and (2) Evaluate the effect of targeted therapy (BRAFV600E, vemurafenib) on the repertoire of melanoma unique antigens. This exploratory study should provide insights into the validity and extent of somatic mutations as unique tumor antigens pave a new strategy to query genomic data for tumor antigen identification and potentially lead to improved patient-specific therapies in melanoma as well as other cancers.

描述（由申请人提供）：在2012年美国，黑色素瘤的发病率在全球范围内继续增加，估计有76,250例新病例。尽管新疗法出现了，但黑色素瘤仍然是无法治愈的恶性肿瘤，因此代表了未满足医疗需求的疾病领域。黑色素瘤以与早期紫外线暴露，癌症基因组中的最高突变率（每MB 14-30）的关系以及诱导自发T细胞免疫的能力而闻名。黑色素瘤中最常见的（60％）的躯体非同义突变是BRAFV600E。针对抑制这种驱动因素突变的靶向疗法，例如vemurafenib，导致了很高的反应率，尽管由于获得了耐药性，但持续时间有限。相比之下，基于免疫的疗法的反应率通常很耐用。新兴数据表明，BRAFV600E通过矛盾的MAPK信号传导抑制，增强了抗肿瘤T细胞免疫。总的来说，这些发现支持测试维美富尼与免疫疗法相结合以改善临床结果。在免疫疗法中，研究疫苗和收养T细胞疗法（ACT）开始在早期临床试验中显示出疗效。我们最近使用CD40L/IFN-的PILOT 1期临床试验？成熟的树突状细胞（DC）和黑素细胞谱系限制的GP100抗原揭示了由DC产生的IL-12的治疗益处，用于疫苗转移性黑色素瘤患者。但是，改善疫苗和ACT的开发的关键障碍是经过验证的黑色素瘤抗原的性质和稀少。迄今为止，针对免疫干预的抗原主要来自结构不变的种系/谱系/分化蛋白，并且表现出有限的临床益处。为了开发下一代个性化的癌症免疫疗法，需要采取新的策略来识别患者特异性/独特的肿瘤抗原。临床前模型中的实验证据支持以下论点：这些独特的肿瘤抗原主要是在肿瘤转化过程中引起的，可以引起能够保护宿主免受癌症进展的T细胞免疫。混合捕获转录组测序的出现（RNA捕获测序，RNA-CAP SEQ）提供了一种敏感的方法，可以询问癌症基因组以表达体细胞突变并评估其表达水平。我们正在进行的研究黑色素瘤转录组的实验发现了500多个表达每个肿瘤的体细胞非同义突变。我们建议使用RNA-CAP SEQ以及Insilico HLA I类肽结合算法和体外T细胞测定法测试以下假说：体细胞非同义突变会导致独特的黑色素瘤抗原以及对BRAFV600E抑制的耐药性伴随着梅拉氏症的变化。该假设将在以下特定目的的实验中解决：（1）表征黑色素瘤表达的体细胞突变，并评估其潜力作为独特的抗原，（2）评估靶向治疗（BRAFV600E，Vemurafenib）对黑素瘤独特抗原的作用。这项探索性研究应提供有关躯体突变的有效性和程度的洞察力，因为独特的肿瘤抗原铺平了一种新的策略，可以将基因组数据查询肿瘤抗原鉴定，并可能导致黑色素瘤以及其他癌症的患者特异性疗法改善。