Single cell mutational analysis of PIK3CA in circulating tumor cells and metastases in breast cancer reveals heterogeneity, discordance, and mutation persistence in cultured disseminated tumor cells from bone marrow.
Single cell mutational analysis of PIK3CA in circulating tumor cells and metastases in breast cancer reveals heterogeneity, discordance, and mutation persistence in cultured disseminated tumor cells from bone marrow.
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DOI：
10.1186/1471-2407-14-456
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发表时间：
2014-06-19
期刊：
影响因子：
3.8
通讯作者：
Jeffrey SS
中科院分区：
文献类型：
--
作者：
Deng G;Krishnakumar S;Powell AA;Zhang H;Mindrinos MN;Telli ML;Davis RW;Jeffrey SS
癌症的治疗决策通常是由分子生物标志物指导的，或者对于一些较新的治疗方法，是由原发肿瘤基因型指导的。然而，由于生物标志物或基因型可能随着新的转移灶的出现而发生变化，人们正在研究血液循环肿瘤细胞（CTCs）在疾病进展过程中指导实时药物选择的作用，期望CTCs能够全面代表转移灶中基因组变化的全部谱。然而，通过单细胞分析识别乳腺癌中ctc和转移的突变异质性的信息有限。骨髓中弥散性肿瘤细胞（DTC）的存在在乳腺癌中也具有预后意义，但CTC和DTC检测之间存在差异。在这里，我们分析了PIK3CA突变的一系列单个肿瘤细胞、CTC和dtc，并报告了CTC和相应的转移基因型。我们使用MagSweeper，一种免疫磁分离设备，从乳腺癌患者的原发和转移组织、血液和骨髓中捕获活的单个肿瘤细胞。在单个细胞中筛选PIK3CA基因外显子9和20的突变。在细胞培养中捕获的dtc也进行了PIK3CA突变测序。从17例患者中分离出242个个体肿瘤细胞并进行突变检测，在3例患者中鉴定出48个突变肿瘤细胞。单细胞分析揭示了组织中ctc和肿瘤细胞之间的突变异质性。在连续随访的患者中，ctc、dtc和转移之间以及不同时间点分离的ctc之间存在突变不一致。该患者体外培养的dtc含有PIK3CA突变，尽管在21天的细胞培养过程中形态学发生了变化，但该突变仍保持不变。ctc的单细胞分析可以显示基因型异质性，随时间的变化，以及dtc和远处转移的不一致性。我们提出了一个值得警惕的病例，表明任何一次抽血的CTC并不总是反映转移性基因型，CTC和DTC分析可能提供独立的临床信息。分离的dtc仍然可以存活，并且可以在培养中繁殖，同时保持其原始突变状态，可能作为未来研究新药治疗的资源。
Therapeutic decisions in cancer are generally guided by molecular biomarkers or, for some newer therapeutics, primary tumor genotype. However, because biomarkers or genotypes may change as new metastases emerge, circulating tumor cells (CTCs) from blood are being investigated for a role in guiding real-time drug selection during disease progression, expecting that CTCs will comprehensively represent the full spectrum of genomic changes in metastases. However, information is limited regarding mutational heterogeneity among CTCs and metastases in breast cancer as discerned by single cell analysis. The presence of disseminated tumor cells (DTCs) in bone marrow also carry prognostic significance in breast cancer, but with variability between CTC and DTC detection. Here we analyze a series of single tumor cells, CTCs, and DTCs for PIK3CA mutations and report CTC and corresponding metastatic genotypes. We used the MagSweeper, an immunomagnetic separation device, to capture live single tumor cells from breast cancer patients’ primary and metastatic tissues, blood, and bone marrow. Single cells were screened for mutations in exons 9 and 20 of the PIK3CA gene. Captured DTCs grown in cell culture were also sequenced for PIK3CA mutations. Among 242 individual tumor cells isolated from 17 patients and tested for mutations, 48 mutated tumor cells were identified in three patients. Single cell analyses revealed mutational heterogeneity among CTCs and tumor cells in tissues. In a patient followed serially, there was mutational discordance between CTCs, DTCs, and metastases, and among CTCs isolated at different time points. DTCs from this patient propagated in vitro contained a PIK3CA mutation, which was maintained despite morphological changes during 21 days of cell culture. Single cell analysis of CTCs can demonstrate genotypic heterogeneity, changes over time, and discordance from DTCs and distant metastases. We present a cautionary case showing that CTCs from any single blood draw do not always reflect metastatic genotype, and that CTC and DTC analyses may provide independent clinical information. Isolated DTCs remain viable and can be propagated in culture while maintaining their original mutational status, potentially serving as a future resource for investigating new drug therapies.
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影响因子：
158.5
作者：
Cristofanilli, M;Budd, GT;Hayes, DF
通讯作者：
Hayes, DF
DOI：
10.1002/pros.22625
发表时间：
2013-06
期刊：
The Prostate
影响因子：
--
作者：
Chen CL;Mahalingam D;Osmulski P;Jadhav RR;Wang CM;Leach RJ;Chang TC;Weitman SD;Kumar AP;Sun L;Gaczynska ME;Thompson IM;Huang TH
通讯作者：
Huang TH
DOI：
10.1158/1078-0432.ccr-13-2332
发表时间：
2014-04-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Jeselsohn R;Yelensky R;Buchwalter G;Frampton G;Meric-Bernstam F;Gonzalez-Angulo AM;Ferrer-Lozano J;Perez-Fidalgo JA;Cristofanilli M;Gómez H;Arteaga CL;Giltnane J;Balko JM;Cronin MT;Jarosz M;Sun J;Hawryluk M;Lipson D;Otto G;Ross JS;Dvir A;Soussan-Gutman L;Wolf I;Rubinek T;Gilmore L;Schnitt S;Come SE;Pusztai L;Stephens P;Brown M;Miller VA
通讯作者：
Miller VA
DOI：
10.1186/bcr2131
发表时间：
2008
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Deng G;Herrler M;Burgess D;Manna E;Krag D;Burke JF
通讯作者：
Burke JF
影响因子：
11.2
作者：
Heitzer, Ellen;Auer, Martina;Speicher, Michael R.
通讯作者：
Speicher, Michael R.