The role of B-cells in the anti-tumor immune response in triple negative breast cancer

项目摘要/摘要 乳腺癌是女性癌症相关死亡的主要原因。三阴性乳腺癌(TNBC)有 在所有主要的乳腺癌亚型中预后最差。免疫治疗的预期成功并没有 在TNBC上看到过。T细胞在TNBC和免疫治疗中的作用已经被很好地研究了，但肿瘤- 浸润性B细胞(TIL-B)则鲜为人知。在高达60%的乳腺癌中，TIL-B与 改善预后和对化疗的反应。有证据表明TIL-B对肿瘤有直接反应 TNBC中的抗原。利用TNBC中肿瘤浸润性淋巴细胞的单细胞测序，我们已经鉴定 高度表达低多样性配对的重链和轻链B细胞受体片段，这是决定性的 克隆选择的证据，表明B细胞已经亲和力成熟，可以识别抗原。 为了克隆抗体并确定其在TNBC中的靶点，我们成功地对B细胞进行了测序 到目前为止，我们已经确认的最丰富的B细胞克隆的受体。临床前数据还表明， B细胞抗体的产生在免疫治疗疗效中的重要作用。基于这些数据，我们假设 TNBC中的B细胞识别肿瘤相关抗原并产生生物学上相关的肿瘤特异性 抗体。为了验证我们的假设，我们提出了以下目标：1)发现克隆选择的B- 人TNBC中的细胞/浆母细胞，2)表征针对TNBC的内源性抗体反应，以及3) 评估免疫检查点抑制对人TNBC中B细胞受体多样性和表型的影响。 这些目标的成功实现将支持B细胞在抗肿瘤免疫反应中的作用，并为 为在TNBC中开发基于B细胞的新疗法和/或生物标记物奠定基础。除了推进 在科学知识方面，这项建议为内科科学家提供了培训。唐斯-坎纳博士是一位实践者 乳腺外科肿瘤学家，有细胞免疫学背景。她的长期目标是将她的临床 和研究专业知识来开发一个独立资助的研究项目，重点是 乳腺癌。她受益于两位久负盛名的导师，他们有培训临床医生和科学家的记录 以及极具支持性的研究和实践环境。为了实现她的长期目标， 候选人的提案包括一份结构化的职业发展计划和以下方面的培训：1)生物信息学2)抗体 克隆技术和专业知识的发展：1)B细胞生物学和2)日益复杂的假说 推动实验设计、执行和分析。提议的计划包括有指导的体验式学习， 课程工作和会议的参与，频繁的导师会议和逐渐增加的研究 独立。在获奖期结束时，候选人将准备申请独立 为她的研究项目提供资金。

PROJECT SUMMARY/ABSTRACT Breast cancer is the leading cause of cancer-related death in women. Triple negative breast cancer (TNBC) has the poorest prognosis of all major subtypes of breast cancer. The anticipated success of immunotherapy has not been seen in TNBC. The role of T-cells in TNBC and immunotherapy has been well-studied but the role of tumor- infiltrating B-cells (TIL-B) is less well-known. Present in up to 60% of breast cancers, TIL-B are associated with improved prognosis and response to chemotherapy. Evidence suggests that TIL-B respond directly to tumor antigens in TNBC. Utilizing single-cell sequencing of tumor-infiltrating lymphocytes in TNBC, we have identified high expression of a low diversity of paired heavy and light chain B-cell receptor segments which is definitive evidence of clonal selection, suggesting B-cells have undergone affinity maturation and can recognize antigens. In order to clone antibodies and identify their targets in TNBC, we have successfully sequenced the B-cell receptors of the most abundant B-cell clones we have identified thus far. Pre-clinical data also suggests an important role for B-cell antibody production in immunotherapy efficacy. Based on these data, we hypothesize that B-cells in TNBC recognize tumor-associated antigens and produce biologically relevant tumor-specific antibodies. To test our hypothesis, we propose the following aims: 1) Discover the role of clonally selected B- cells/plasmablasts in human TNBC, 2) Characterize the endogenous antibody response against TNBC, and 3) Evaluate the impact of immune checkpoint inhibition on B-cell receptor diversity and phenotype in human TNBC. Successful execution of these aims will support a role for B-cells in the anti-tumor immune response and lay the foundation for development of novel B-cell based therapies and/or biomarkers in TNBC. In addition to advancing scientific knowledge, this proposal provides training to a physician-scientist. Dr. Downs-Canner is a practicing breast-surgical oncologist with a background in cellular immunology. Her long-term goal is to combine her clinical and research expertise to develop an independently-funded research program focused on immunotherapy in breast cancer. She benefits from two well-established mentors with track records of training clinician-scientists and an extremely supportive research and practice environment. In order to achieve her long-term goals, the candidate's proposal includes a structured career development plan and training in: 1) bioinformatics 2) antibody cloning techniques and development of expertise in: 1) B-cell biology and 2) increasingly complex hypothesis driven experimental design, execution, and analysis. The proposed plan includes mentored experiential learning, course work and conference participation, frequent mentor meetings and a gradual increase in research independence. At the completion of the award period, the candidate will be prepared to apply for independent funding for her research program.