Immunomagnetic Enrichment, Genomic Characterization, and Prognostic Impact of Circulating Melanoma Cells

Ulmer, Anja; Schmidt‐Kittler, Oleg; Fischer, Jörg; Ellwanger, Ulf; Rassner, G.; Riethmüller, Gert; Fierlbeck, Gerhard

doi:10.1158/1078-0432.ccr-0424-03

Cited by 111 publications

(103 citation statements)

References 21 publications

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“…Patients who undergo resection of lymph node basins harboring metastasis do not experience a significantly extended life expectancy (3,4). Furthermore, circulating melanoma cells were detected in the blood of 26% of patients who only have metastases detected regionally (5,6).…”

mentioning

confidence: 99%

Phylogenetic analyses of melanoma reveal complex patterns of metastatic dissemination

Sanborn¹,

Chung

Purdom

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

156

133

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Melanoma is difficult to treat once it becomes metastatic. However, the precise ancestral relationship between primary tumors and their metastases is not well understood. We performed whole-exome sequencing of primary melanomas and multiple matched metastases from eight patients to elucidate their phylogenetic relationships. In six of eight patients, we found that genetically distinct cell populations in the primary tumor metastasized in parallel to different anatomic sites, rather than sequentially from one site to the next. In five of these six patients, the metastasizing cells had themselves arisen from a common parental subpopulation in the primary, indicating that the ability to establish metastases is a late-evolving trait. Interestingly, we discovered that individual metastases were sometimes founded by multiple cell populations of the primary that were genetically distinct. Such establishment of metastases by multiple tumor subpopulations could help explain why identical resistance variants are identified in different sites after initial response to systemic therapy. One primary tumor harbored two subclones with different oncogenic mutations in CTNNB1, which were both propagated to the same metastasis, raising the possibility that activation of wingless-type mouse mammary tumor virus integration site (WNT) signaling may be involved, as has been suggested by experimental models.A s in many other solid tumors, melanoma metastases often first present in lymph nodes in the draining area of the primary, whereas distant metastases tend to appear later (1). The conclusion that melanoma follows a linear progression from primary tumor to regional to distant metastases has supported preemptive surgical removal of regional lymph nodes with curative intent (2). However, several observations suggest that distant metastases are seeded early, contemporaneously with regional metastases. Patients who undergo resection of lymph node basins harboring metastasis do not experience a significantly extended life expectancy (3, 4). Furthermore, circulating melanoma cells were detected in the blood of 26% of patients who only have metastases detected regionally (5, 6).Melanoma, like other cancers, arises and evolves through the accumulation of genetic alterations within tumor cells (7-9). Comparing somatic mutations in primary tumor and regional and distant metastases from the same patient can provide insight into the phylogenetic relationships between these distinct tumor cell populations and the order of metastatic dissemination (8, 10). These analyses may also establish whether cells in the primary tumor that metastasize acquired this ability to disseminate and seed other anatomic sites by a newly acquired genetic alteration, or whether metastatic colonization is simply a stochastic process of which all cells in the primary are capable but few succeed.Using whole-exome sequencing (for discovery) and targeted sequencing (for validation), we analyzed mutation patterns of primary melanomas and two or more metastases in each of e...

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mentioning

confidence: 99%

Phylogenetic analyses of melanoma reveal complex patterns of metastatic dissemination

Sanborn¹,

Chung

Purdom

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

156

133

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“…From these results it is obvious that; a) CTCs are present at relatively low concentrations; one tumour cell per 10 6 to 10 7 normal blood cells or on average, 1 cell per ml of blood 69,70,71 ; b) the number of cells appears to be related to stage 2,25,72 ; c) melanoma cell markers differ with respect to stage 73 ; and c) CTC gene expression differs from that of the primary tumour 28,29 . Quantitative RT-PCR has typically detected expression of melanocytic genes such as tyrosinase (TYR) 74,75 since normal melanocytes are not thought to circulate in peripheral blood and therefore detection of transcripts from melanocytic genes should correlate to identification of CTCs 72,76 .…”

Section: Detection Of Ctcsmentioning

confidence: 93%

“…Various methods have been used to quantify and characterise CTCs, including indirect methods, namely qRT-PCR 25,[62][63][64][65] , and direct analyses such as immunomagnetic bead capture, or fibre-optic array scanning technology 28,29,66,67,68 . From these results it is obvious that; a) CTCs are present at relatively low concentrations; one tumour cell per 10 6 to 10 7 normal blood cells or on average, 1 cell per ml of blood 69,70,71 ; b) the number of cells appears to be related to stage 2,25,72 ; c) melanoma cell markers differ with respect to stage 73 ; and c) CTC gene expression differs from that of the primary tumour 28,29 .…”

Section: Detection Of Ctcsmentioning

confidence: 99%

“…Several studies have shown that the number of CTCs in patient peripheral blood increases with increasing stages of melanoma 25,[26][27][28][29]30 . Yet the variety and phenotype of these CTCs, their ability to remain in circulation for many years thus evading the immune system, and their activation causing metastasis, remain largely unexplored, particularly for melanoma.…”

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confidence: 99%

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Circulating Melanoma Cells

Ziman¹

2011

Breakthroughs in Melanoma Research

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“…One approach to cancer cell enrichment has been the use of immunomagnetic systems, in which antibodies against epithelial surface markers are linked to small paramagnetic microbeads, enabling target-cell selection using a powerful magnet (2)(3)(4)(5). Many researchers have successfully used such systems for early detection of recurrent carcinomas, mainly using blood samples.…”

Section: Introductionmentioning

confidence: 99%

Diagnostic potential and limitation of imaging cancer cells in cytological samples using telomerase-specific replicative adenovirus

et al. 2009

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Abstract. Cytological cancer screening that targets genetic or epigenetic abnormalities may be a viable alternative to morphological screening. Detecting cancer cells by specific genetic markers helps their easy detection in cytological samples. We recently established the telomerase-specific replication-selective adenovirus OBP-401, in which the human telomerase reverse transcriptase (hTERT) gene promoter has been inserted upstream of the E1 genes, and in which the green fluorescent protein (GFP) gene is driven by the CMV promoter. This virus selectively replicates only in telomerasepositive cells, expressing GFP, and therefore may be a tool for cancer screening. In the present study, we first confirmed that cytological samples can easily be infected with OBP-401, allowing visualization of GFP-positive cells under fluorescent microscopy 24 h after infection. After 32 cytological samples from patients with cervical, endometrial or ovarian cancers were infected with OBP-401, GFP signals were detected in 31 (96%) of the samples. However, some normal endometrial scrapings exhibited GFP-signals, possibly due to endometrial glandular cells with constitutive telomerase activity. The ability of OBP-401 to enrich cancer cells was then tested. Cytological samples containing cervical or endometrial cancer cells were infected with OBP-401, and GFP-positive cells were sorted by flow cytometry; DNA was extracted from the GFP-positive cells. Direct DNA sequencing or methylationspecific PCR identified cancer-derived mutations or hypermethylations of tumor suppressor genes more efficiently than analyses using crude cytological samples. Thus, OBP-401-based sorting of GFP-positive cells successfully enriched cancer cells, allowing efficient detection of genetic or epigenetic abnormalities in cytological samples.

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Immunomagnetic Enrichment, Genomic Characterization, and Prognostic Impact of Circulating Melanoma Cells

Cited by 111 publications

References 21 publications

Phylogenetic analyses of melanoma reveal complex patterns of metastatic dissemination

Phylogenetic analyses of melanoma reveal complex patterns of metastatic dissemination

Circulating Melanoma Cells

Diagnostic potential and limitation of imaging cancer cells in cytological samples using telomerase-specific replicative adenovirus

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