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小木蟲劉樹青木蟲 (著名寫手)
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potential for using CTCs for high-throughput molecular analyses. Beyond studies evaluating the clinical utility of CTC enumeration and the characterization of a limited number of markers, we anticipate that in the future CTCs might be evaluated as a tissue source in molecular screening programs. Several groups have demonstrated the feasibility of analyzing CTC-enriched fractions [58] or pure CTCs [59] for the expression of a number of preselected transcripts. These studies have revealed a profound heterogeneity of CTCs at the transcriptional level. In addition to characterizing gene expression, efforts have also been made to study the mutational profiles of CTCs in various cancer types. A recent study has analyzed CTCs isolated by CellSearch®, primary tumor and metastases from six metastatic colorectal cancer patients using array comparative genomic hybridization and NGS for a panel of 68 cancer genes. They observed that most mutations initially found only on CTCs were also present as subclonal mutations in primary tumors and metastases. We have also demonstrated the feasibility of performing mutational analysis on CTCs detected by CellSearch® and purified using DEPArray™[61]. Recently,investigators showed the feasibility of CTC whole-exome sequencing and reliable somatic single-nucleotide variant (SNV) calling in patients with metastatic prostate cancer and at least 10 CTCs/7.5 ml of blood. They have employed a modular approach consisting of CTC isolation with the Magsweeper technology, single-cell whole-genome amplification, library qualification and a census-based sequencing strategy [63]. This approach allows for confident variant calling when the variant is present in many CTC libraries from the same sample allowing for the identification of truncal mutations but cannot be used to study heterogeneity between single CTCs. The above studies demonstrate the feasibility of performing genomic and transcriptomic analysis on CTCs. Advances in single-cell sequencing [64–66] and the application of these technologies to serial CTC analyses have significant potential to improve our understanding of heterogeneity and disease evolution. Beyond such analyses, there is now evidence that CTCs can be used as a tissue source for drug sensitivity testing. Indeed, investigators were able to culture CTCs ex vivo and identified activating ESR1 mutations in three of the six CTC-derived cell lines from aromatase inhibitor-pretreated, ER-positive, MBC patients. These mutations are very rarely observed in primary or treatment-naïve ER-breast cancer but were recently suggested to occur at a frequency of 15% in hormone-resistant breast cancer.Using these CTC-derived cell lines, they confirmed previousfindings that ESR1-mutant cells were resistant to tamoxifen, raloxifene and fulvestrant and provided novel evidence for antitumor activity using a combination of raloxifene or fulvestrant with an HSP90 inhibitor [67]. The same group carried out drug sensitivity testing in a mouse xenograft model from a CTC cell line harboring a PIK3CA and an FGFR2 mutation. Similarly, other groups have shown that CTCs from patients with breast [70] and small-cell lung cancer (SCLC) [71] were tumorigenic in mouse models. Interestingly, CTC-derived, SCLC mouse models demonstrated similar patterns of response to platinum-based chemotherapy as the original donor-patients. The clinical utility of the CTC models (CTC-derived cell lines or mouse xenografts) will depend on (i) the percentage of patients in whom this approach is feasible and (ii) whether theCTC models can always reliably capture response to different drugs. In the future, we could imagine that CTC genomic and transcriptomic analysis together with drug sensitivity testing in CTC-derived cell lines and mouse models might guide personalized treatment approaches. |
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2樓2016-03-27 08:52:20
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