DEVELOPMENT OF GENOMICS BASED PERSONALIZED CANCER IMMUNOTHERAPY

 描述（由申请人提供）：2012年，我们描述了下一代测序和生物信息学方法的首批用途之一，用于快速准确地鉴定肿瘤特异性突变蛋白，该蛋白在高度免疫原性（未经编辑）、化学诱导的小鼠肿瘤系中充当主要排斥抗原，该小鼠肿瘤系在移植到幼稚同基因野生型小鼠中时被排斥。自从发表这项工作以来，我们已经产生了令人信服的初步数据，使我们形成了这样的假设，即在成功的检查点阻断癌症免疫治疗期间刺激的CD 8 + T细胞识别的临床明显（编辑）肿瘤细胞中的抗原也可能是肿瘤特异性突变蛋白，并且它们也可以使用我们的基因组学方法快速识别。在这个提案中，我们希望正式测试这一假设，为我们的方法最终转化为人类癌症患者提供基础，以促进癌症免疫治疗的个性化。为了实现这一目标，我们将努力实现三个具体目标。具体目标一：鉴定肿瘤特异性突变抗原，其激发CD 8 + T细胞对MCA肉瘤和B16-F10黑色素瘤的应答，并对检查点阻断疗法具有不同的敏感性。 在这里，我们将集中我们的努力进一步验证，甚至提高我们的能力，以确定那些来自肿瘤特异性突变蛋白，在癌症中有潜在的治疗效用的抗原。我们将询问肿瘤特异性突变蛋白是否是检查点阻断治疗的首选靶点，以及检查点阻断是否仅选择这些突变中最具抗原性的突变。具体目标二。确定靶向肿瘤特异性突变抗原的疫苗，无论是单独使用还是与检查点阻断剂联合使用，是否可以治疗性控制MCA肉瘤或B16-F10黑色素瘤的生长。 在这里，我们将探讨我们在Aim I中鉴定的肿瘤特异性突变抗原是否可以用作治疗性肿瘤特异性癌症疫苗的基础。我们将调查以下三个问题：个性化疫苗能否：（a）代替检查站封锁？(b)在敏感肿瘤中改善检查点阻断？(c)在不敏感肿瘤中引起检查点阻断的有效性？具体目标三。定义肿瘤特异性CD 8 + T细胞的特征，以确定其治疗效果。 这些实验将试图确定激活的肿瘤抗原特异性CD 8 + T细胞的特征，这些特征导致成功的癌症免疫治疗。我们想检验这样一个假设，即CyTOF和RNA-Seq分析将有助于鉴定表达特异性表型标志物的肿瘤特异性CD 8 + T细胞，这些标志物可以预测有利的临床结果。

 DESCRIPTION (provided by applicant): In 2012, we described one of the first uses of next-generation sequencing and bioinformatics approaches to rapidly and accurately identify a tumor-specific mutant protein that functioned as a major rejection antigen in a highly immunogenic (unedited), chemically induced mouse tumor line that was rejected when transplanted into naïve syngeneic wild type mice. Since publishing this work, we have generated compelling preliminary data leading us to form the hypothesis that the antigens in clinically apparent (edited) tumor cells that are recognized by CD8+ T cells stimulated during successful checkpoint blockade cancer immunotherapy may also be tumor-specific mutant proteins and that they too can be rapidly identified using our genomics approach. In this proposal, we wish to formally test this hypothesis to provide a foundation for the eventual translation of our method to human cancer patients so as to facilitate personalization of cancer immunotherapies. To achieve this goal we will pursue three specific aims. Specific Aim I: Identify Tumor-specific Mutational Antigens Eliciting CD8+ T Cell Responses to MCA Sarcomas and B16-F10 Melanoma With Differential Sensitivities to Checkpoint Blockade Therapy. Here we will focus our efforts on further validating and perhaps even improving our capacity to identify those antigens derived from tumor-specific mutant proteins that have potential therapeutic utility in cancer. We will ask whether tumor-specific mutant proteins are the favored targets of checkpoint blockade therapy and whether checkpoint blockade selects only the most antigenic of these mutations. Specific Aim II. Determine Whether Vaccines Targeting Tumor-Specific Mutational Antigens, Either Alone or in Combination with Checkpoint Blockade, Can Therapeutically Control Growth of MCA Sarcomas or B16-F10 Melanoma. Here we will explore whether the tumor-specific mutant antigens we identify in Aim I can be used as a basis for therapeutic tumor-specific cancer vaccines. We will investigate the following three questions: Can personalized vaccines: (a) be used instead of checkpoint blockade? (b) improve checkpoint blockade in sensitive tumors? (c) evoke checkpoint blockade effectiveness in insensitive tumors? Specific Aim III. Define the Characteristics of Tumor-specific CD8+ T Cells that Specify Their Therapeutic Effectiveness. These experiments will seek to define the characteristics of activated tumor antigen-specific CD8+ T cells that result in successful cancer immunotherapy. We want to test the hypothesis that CyTOF and RNA-Seq analyses will facilitate identification of tumor-specific CD8+ T cells expressing specific phenotypic markers that can predict a favorable clinical outcome.