Somatic mutations in leukocytes infiltrating primary breast cancers.
Somatic mutations in leukocytes infiltrating primary breast cancers.
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DOI：
10.1038/npjbcancer.2015.5
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发表时间：
2015
影响因子：
5.9
通讯作者：
Norton L
中科院分区：
文献类型：
--
作者：
Kleppe M;Comen E;Wen HY;Bastian L;Blum B;Rapaport FT;Keller M;Granot Z;Socci N;Viale A;You D;Benezra R;Weigelt B;Brogi E;Berger MF;Reis-Filho JS;Levine RL;Norton L
癌变需要癌细胞与其微环境的相互作用，包括渗透的白细胞。然而，体细胞突变研究的重点是癌细胞的变化，假设微环境在遗传上是正常的。因为我们假设这可能不是一个有效的假设，所以我们进行了外显子组测序和靶向测序，以调查乳腺癌中肿瘤相关白细胞中致病突变的存在。我们使用靶向测序和外显子组测序来评估从未经治疗的原发乳腺癌中分离的肿瘤浸润性CD45阳性细胞中是否存在突变。我们使用高深度测序来确定我们在乳腺癌浸润性白细胞、纯化的肿瘤细胞和循环血细胞中发现的突变的存在/不存在。对15对肿瘤浸润性白细胞和配对的种系DNA进行捕获测序，发现在我们队列中所有15名原发性乳腺癌患者的已知癌症基因中存在变异。我们通过正交外显子测序验证了通过靶向测序在浸润性白细胞中发现的突变的存在。10名患者存在先前报道的躯体获得性变异，包括已知的白血病基因(DNTM3A、TET2和BCOR)。在肿瘤浸润性白细胞中观察到的其中一个突变在同一患者的循环白细胞中也被检测到，其等位基因频率低于肿瘤浸润性细胞中的等位基因频率。在这里，我们展示了体细胞突变，包括已知癌症基因的突变，存在于浸润性原发乳腺癌亚群的白细胞中。这一观察允许癌细胞与突变型浸润性白细胞相互作用的可能性，这具有许多潜在的临床意义。
Malignant transformation requires the interaction of cancer cells with their microenvironment, including infiltrating leukocytes. However, somatic mutational studies have focused on alterations in cancer cells, assuming that the microenvironment is genetically normal. Because we hypothesized that this might not be a valid assumption, we performed exome sequencing and targeted sequencing to investigate for the presence of pathogenic mutations in tumor-associated leukocytes in breast cancers. We used targeted sequencing and exome sequencing to evaluate the presence of mutations in sorted tumor-infiltrating CD45-positive cells from primary untreated breast cancers. We used high-depth sequencing to determine the presence/absence of the mutations we identified in breast cancer-infiltrating leukocytes in purified tumor cells and in circulating blood cells. Capture-based sequencing of 15 paired tumor-infiltrating leukocytes and matched germline DNA identified variants in known cancer genes in all 15 primary breast cancer patients in our cohort. We validated the presence of mutations identified by targeted sequencing in infiltrating leukocytes through orthogonal exome sequencing. Ten patients harbored alterations previously reported as somatically acquired variants, including in known leukemia genes (DNTM3A, TET2, and BCOR). One of the mutations observed in the tumor-infiltrating leukocytes was also detected in the circulating leukocytes of the same patients at a lower allele frequency than observed in the tumor-infiltrating cells. Here we show that somatic mutations, including mutations in known cancer genes, are present in the leukocytes infiltrating a subset of primary breast cancers. This observation allows for the possibility that the cancer cells interact with mutant infiltrating leukocytes, which has many potential clinical implications.