Evolution and clinical impact of clonal hematopoiesis of indeterminate potential in breast tumor microenvironment

项目概要/摘要 不确定潜能的克隆性造血（ClonalHematopoiesis ofIndeterminatePotential，CHIP）是一种与年龄相关的造血干细胞扩增 具有体细胞改变而不呈现其他血液学异常的细胞。芯片已经 在患有实体瘤的癌症患者的正常外周血中检测到，并且已经被认为具有 在治疗相关的继发性骨髓疾病发展中的容许作用。我们最近的临床分析 对113，079个实体瘤标本的测序表明，CHIP克隆也存在于实体瘤中。 肿瘤微环境由于混合突变造血元素;然而，富集混合 CHIP、其在治疗下的演变及其对实体瘤治疗的临床影响知之甚少。 在这个建议中，我们将在乳腺浸润性癌的背景下描述CHIP， 化疗药物在辅助和新辅助环境中的应用，并已显示具有预后 与微环境中的浸润造血细胞相互作用。我们假设CHIP表现出 当在乳腺肿瘤微环境中富集时， 治疗，并与治疗诱导的血液学毒性的发展相关。测试这些 假设，我们将收集1,200名新诊断的乳腺癌患者的队列， 全面的临床数据以及治疗前后的外周血和乳腺肿瘤 样品，并在三个目标的高分辨率CHIP配置文件。首先，我们将确定 在乳腺肿瘤治疗之前混合CHIP。使用外周血和乳腺癌的高深度测序 在肿瘤样本中，我们将检测>0.1%等位基因频率的CHIP突变，并将 将CHIP与浸润淋巴细胞和其他造血标志物的水平混合。使用单细胞 通过基因组分析，我们将解决专属CHIP克隆的数量。第二，我们将分析芯片后， 外周血和乳腺肿瘤样本中的新辅助治疗，以研究其演变 通过评估突变特异性适应性和治疗瓶颈大小，使用激素治疗作为对照。我们 还将研究粒细胞集落刺激对CHIP克隆动力学的影响。纵向 外周血采样将阐明CHIP的长期演变结束后1-2年的乳房 肿瘤的治疗。第三，我们将开发一个统计回归模型来确定不同的CHIP克隆 这可能与临床反应和治疗诱导的血液学毒性的发生相关。 这项研究是新颖的，它利用了系统收集的临床和高分辨率分子数据， 这将为研究CHIP在乳腺肿瘤治疗中的克隆进化提供新的思路。此外，它将说明 造血细胞群体分子克隆分析作为基本预测指标的意义 治疗引起的血液学并发症最后，它将建立一个长期的临床和 高危患者CHIP进展为治疗相关髓系肿瘤的分子研究。

PROJECT SUMMARY/ABSTRACT Clonal hematopoiesis of indeterminate potential (CHIP) is an age-related expansion of hematopoietic stem cells that harbor somatic alterations without presenting other hematologic abnormalities. CHIP has been detected in normal peripheral blood of cancer patients with solid tumors and has been suggested to have a permissive role in therapy-related secondary myeloid disease development. Our recent analysis of clinical sequencing of 113,079 solid tumor specimens demonstrated that CHIP clones are also present in the solid tumor microenvironment due to admixed mutated hematopoietic elements; however, enrichment of admixed CHIP, its evolution under therapy, and its clinical impact on solid tumor treatment are poorly understood. In this proposal, we will characterize CHIP in the context of breast invasive carcinoma, which is treated with chemotherapeutics in both adjuvant and neoadjuvant settings and has been shown to have prognostic interactions with infiltrating hematopoietic cells in its microenvironment. We hypothesize that CHIP exhibits a distinct genomic landscape when enriched in breast tumor microenvironment, evolves under breast tumor treatment, and is correlated with the development of therapy-induced hematological toxicity. To test these hypotheses, we will assemble a cohort of 1,200 newly diagnosed breast cancer patients, collect comprehensive clinical data as well as sequential pre- and post-treatment peripheral blood and breast tumor samples, and profile CHIP at high resolution in three aims. First, we will determine the mutational spectrum of admixed CHIP before breast tumor treatment. Using high-depth sequencing of peripheral blood and breast tumor samples, we will detect CHIP mutations at >0.1% allele frequency and correlate the prevalence of admixed CHIP with the level of infiltrating lymphocytes and other hematopoietic markers. Using single-cell genomic analysis, we will resolve the number of exclusive CHIP clones. Second, we will profile CHIP after chemotherapy in peripheral blood – and breast tumor samples in neoadjuvant settings – to study its evolution by assessing mutation-specific fitness and therapeutic bottleneck size, using hormonal therapy as control. We will also investigate the effect of granulocyte-colony stimulation on CHIP's clonal dynamics. Longitudinal peripheral blood sampling will elucidate the long-term evolution of CHIP 1-2 years after the end of breast tumor's treatment. Third, we will develop a statistical regression model to determine the distinct CHIP clones that may be correlated with clinical response and development of therapy-induced hematological toxicity. This study is novel in its utilization of systematically collected clinical and high-resolution molecular data, and it will provide insight on CHIP's clonal evolution under breast tumor treatment. Moreover, it will illustrate the significance of molecularly defined clonal analysis of hematopoietic populations as a fundamental predictor of therapy-induced hematological complications. Finally, it will establish a platform for long-term clinical and molecular inquiries of CHIP's progression to therapy-related myeloid neoplasms in high-risk patients.