Serial RT-PCR detection of circulating tumour cells as a marker of disease progression in patients with malignant melanoma

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 18009-18009
Author(s):  
P. A. Ascierto ◽  
M. Budroni ◽  
A. Cossu ◽  
S. Scala ◽  
E. Simeone ◽  
...  
Keyword(s):  
Overall Survival ◽  
Malignant Melanoma ◽  
Disease Progression ◽  
Peripheral Blood ◽  
Disease Stage ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Blood Samples ◽  
Melanoma Patients

18009 Background: Detection of circulating malignant cells (CMCs) through a reverse transcriptase-polymerase chain reaction (RT-PCR) assay seems to be a demonstration of systemic disease. We here evaluated the prognostic role of RT-PCR assays in serially-taken peripheral blood samples from patients with malignant melanoma (MM). Methods: One hundred forty-nine melanoma patients with disease stage ranging from I to III were consecutively collected in 1997. A multi-marker RT-PCR assay was used on peripheral blood samples obtained at time of diagnosis and every 6 months during the first two years of follow-up (total: 5 samples). Univariate and multivariate analyses were performed after 83 months of median follow-up. Results: Detection of at least one circulating mRNA marker was considered a signal of the presence of CMC (referred to as PCR-positive assay). A significant correlation was found between the rate of recurrences and the increasing number of PCR-positive assays (P = 0.007). Presence of CMC in a high number (≥2) of analysed blood samples was significantly correlated with a poor clinical outcome (disease-free survival: P = 0.019; overall survival: P = 0.034). Multivariate analysis revealed that presence of a PCR-positive status does play a role as independent prognostic factors for overall survival in melanoma patients, adding precision to the predictive power of the disease stage. Conclusions: Our findings indicated that serial RT-PCR assay may identify a high risk subset of melanoma patients with occult cancer cells constantly detected in blood circulation. Prolonged presence of CMCs seems to act as a surrogate marker of disease progression or a sign of more aggressive disease. No significant financial relationships to disclose.

Cyclin D1 mRNA Expression In CLL: a Pilot Study.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4619-4619
Author(s):  
Heidi Mocikova ◽  
Sona Pekova ◽  
Lenka Zejskova ◽  
Martin Spacek ◽  
Tomas Kozak
Keyword(s):  
Bone Marrow ◽  
Overall Survival ◽  
Prognostic Factors ◽  
Cyclin D1 ◽  
Mrna Expression ◽  
Peripheral Blood ◽  
Newly Diagnosed ◽  
Rt Pcr ◽  
Trisomy 12

Abstract Abstract 4619 Background. Expression of cyclin D1 demonstrated by immunohistochemistry is seen in 95% of mantle cell lymphomas and in other lymphoproliferative diseases including 10% of chronic lymphocytic leukemias (CLL). This study analyzed the impact of cyclin D1 positivity on time to treatment (TTT) and overall survival (OS) measured by RT PCR in newly diagnosed CLL patients and correlation with reported prognostic factors. Patients and methods. Level of cyclin D1 (quantitative real-time RT-PCR with a specific TaqMan flurescent hybridization probe) mRNA expression was analysed in 72 samples (57 peripheral blood and 15 bone marrow) from patients with newly diagnosed CLL. Cyclin D1 expression (cut-off according to ROC curve >3 over the threshold expression in healthy donors) was reported as positive. Fisher's exact test was used to analyze the relationship of cyclin D1 positivity and prognostic factors: del17p, del11q, unmutated IgVH, trisomy 12, ZAP 70 and CD38 positivity, elevated B2microglobulin, elevated LDH and lymphocyte doubling time (LDT) <6 months. The comparison of time to treatment (TTT) and prognostic factors with cyclin D1 positivity was calculated via Spearman correlation coefficient. Survival curves were calculated by Kaplan-Meier survival analysis and comparison between subgroups was performed by the log-rank test. Results. Cyclin D1 was positive in 29 (40%) CLL patients. Although cyclin D1 was not statistically significant for TTT (P=0,145), a trend was observed, suggesting a negative prognostic impact of cyclin D1 overexpression in CLL. Following variables correlated significantly with TTT: del17p (P=0.037), del11q (P=0.003), unmutated IgVH (P=0.004), trisomy 12(P=0.024), positive CD38 (P=0.014), elevated B2microglobulin (P<0.001), elevated LDH (P<0.001) and LDT <6 months (P<0.001). Del 17p, del 11q, trisomy 12 and elevated B2microglobulin were independent factors for TTT in the multivariate analysis. None of these factors were significant for overall survival due to the short follow-up. Conclusion. Cyclin D1 measured by RT-PCR from peripheral blood or bone marrow has no statistically singificant impact on TTT or OS, though a trend pointing to cyclin D1 overexpression in CLL as a negative prognostic marker can be suggested. This data should be confirmed on a larger cohort of patients with a longer follow-up. Disclosures: No relevant conflicts of interest to declare.

Molecular Staging in Stage II and III Melanoma Patients and Its Effect on Long-Term Survival

2005 ◽  
Vol 23 (6) ◽  
pp. 1218-1227 ◽  
Author(s):  
Christiane Voit ◽  
Martina Kron ◽  
Juergen Rademaker ◽  
Markus Schwürzer-Voit ◽  
Wolfram Sterry ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Prognostic Value ◽  
Stage Ii ◽  
Disease Specific Survival ◽  
Rt Pcr ◽  
Term Survival ◽  
Histologic Subtype ◽  
Melanoma Patients ◽  
Disease Specific

Purpose To assess the prognostic value of serial reverse transcriptase polymerase chain reaction (RT-PCR) -based measurements of tyrosinase mRNA in peripheral blood of stage II and III melanoma patients. Patients and Methods During routine follow-up of American Joint Committee on Cancer stage II and III melanoma patients, serial testing for tyrosinase transcripts in peripheral blood was performed by RT-PCR. The PCR results were compared with the clinical data collected during the follow-up. Results Over a period of 3 years, 111 patients (78 stage II and 33 stage III patients) were enrolled, and tyrosinase determinations were carried out. The 6-year disease-specific survival probability was 97% for patients always showing negative RT-PCR results and 67% for patients who tested positive at least once. In a Cox proportional hazards model, the prognostic value of sex, age, site of primary tumor, histologic subtype, stage, Breslow's tumor thickness, Clark level, and the time-dependent variable PCR result was assessed. Patients with a positive RT-PCR test had a distinctly higher risk of dying from melanoma, with a hazard ratio of 12.6 (95% CI, 3.4 to 46.3; P < .001). Conclusion Our study shows a strong association between PCR and disease-specific survival time. Detection of tyrosinase mRNA in peripheral blood may be of similar importance for the clinical course of melanoma as the detection of micrometastatic disease in the sentinel lymph node. Whether a combination of these two factors leads to a better definition of the prognosis of melanoma patients is under investigation in current studies.

Characteristics and probability of survival for patients with advanced melanoma who live five or more years after initial treatment with immune checkpoint blockade (ICB).

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. 9534-9534
Author(s):  
Kimberly Loo ◽  
Debra A. Goldman ◽  
Katherine Panageas ◽  
Margaret K. Callahan ◽  
Paul B. Chapman ◽  
...  
Keyword(s):  
Overall Survival ◽  
Treatment Failure ◽  
Disease Progression ◽  
Stage Iii ◽  
Late Relapse ◽  
Unresectable Stage ◽  
Melanoma Patients ◽  
Long Term Survivors

9534 Background: A subset of melanoma patients treated with ICB (ipilimumab [ipi], nivolumab [nivo], pembrolizumab [pembro] or nivo+ipi) will experience durable responses. While five-year survival rates have been reported for patients treated with ICB on clinical trials, little is known about the clinical characteristics, survival past five years, and patterns of late relapse of long-term survivors. Methods: We retrospectively reviewed all patients treated at Memorial Sloan Kettering for unresectable stage III/IV melanoma who survived at least five years following their first dose of ICB (N = 151). Demographics, disease characteristics, and nature of progression were examined. Overall survival (OS) was calculated from 5 years post-ICB. Time to Treatment failure (TTF) was calculated conditionally from 5 years out until next therapy, progression, or death. Results: Of the 151 long-term survivors, median age at first ICB treatment was 62 years (range 22-83), with 101 (66.9%) male and 50 (33.1%) female patients. Stage at first ICB treatment was unresectable stage III (26, 17.2%), M1a (21,13.9%), M1b (39, 25.8%), M1c (52, 34.4%), M1d (13, 8.6%). Melanoma subtype was cutaneous (122, 80.8%), unknown primary (24, 15.9%), mucosal (3, 2%), and acral (2, 1.3%). First ICB was ipi (108, 71.5%), PD-1 (nivo or pembro) (5, 3.3%), and nivo+ipi (37, 24.5%). The best overall response to first ICB was CR (76, 50.3%), PR (27, 17.9%), SD (16, 10.6%) and PD (32, 21.2%). Of the patients who progressed after initial ICB, 38 received subsequent systemic treatment as follows: PD-(L)1 in 20 (53%), BRAF ± MEK in 9 (23.7%), ipi in 7 (18.4%), and chemotherapy in 2 (5.3%). Median duration of follow-up among survivors (N = 138) was 93 months (range 60-192). From 5 years post-ICB, 85% (95% CI: 73-92%) survived an additional 5 years. In those who made it to 5 years without treatment failure (N = 72), the probability of remaining failure-free was 92% (95% CI: 86-99%) at 7 years. Of the 151 patients, only 4 patients (2.6%) experienced disease progression after 5 years. Three patients had radiographic or pathologic disease progression in the lymph nodes and one in the subcutaneous tissue. No patients progressed in the lungs, visceral organs, or CNS after 5 years. At time of analysis, 13 (8.6%) patients died after 5 years post ICB, none died of progressive melanoma. 6 patients died of unknown causes, 2 died of other causes, and 5 died of other non-melanoma cancer-related causes. Conclusions: Patients who survive five years after their initial immunotherapy have excellent overall survival and treatment failure-free survival. Given the anxiety surrounding survivorship and late progression, long-term survivors should be reassured of their excellent prognosis. These data suggest that aggressive follow-up schedules and imaging of melanoma patients after 5 years of survival may not be required.

scholarly journals Serum CEACAM1 Correlates with Disease Progression and Survival in Malignant Melanoma Patients

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Sapoznik Sivan ◽  
Faranesh Suzan ◽  
Ortenberg Rona ◽  
Hamburger Tamar ◽  
Barak Vivian ◽  
...  
Keyword(s):  
Immune Response ◽  
Overall Survival ◽  
Malignant Melanoma ◽  
Metastatic Melanoma ◽  
Disease Progression ◽  
Skin Test ◽  
Metastatic Disease ◽  
Prognostic Value ◽  
Serum Samples ◽  
Melanoma Patients

The search for melanoma biomarkers is crucial, as the incidence of melanoma continues to rise. We have previously demonstrated that serum CEACAM1 (sCEACAM1) is secreted from melanoma cells and correlates with disease progression in metastatic melanoma patients. Here, we have used a different cohort of melanoma patients with regional or metastatic disease (N=49), treated with autologous vaccination. By monitoring sCEACAM1 in serum samples obtained prior to and after vaccination, we show that sCEACAM1 correlates with disease state, overall survival, and S100B. The trend of change in sCEACAM1 following vaccination (increase/decrease) inversely correlates with overall survival. DTH skin test is used to evaluate patients’ anti-melanoma immune response and to predict response to vaccination. Importantly, sCEACAM1 had a stronger prognostic value than that of DTH, and when sCEACAM1 decreased following treatment, this was the dominant predictor of increased survival. Collectively, our results point out the relevance of sCEACAM1 in monitoring melanoma patients.

scholarly journals Quantification of Melanoma Cell-specific MART-1 mRNA in Peripheral Blood by a Calibrated Competitive Reverse Transcription-PCR

2000 ◽  
Vol 46 (12) ◽  
pp. 1923-1928 ◽  
Author(s):  
Boe Sandahl Sørensen ◽  
Henrik Schmidt ◽  
Hans von der Maase ◽  
Per Thor Straten ◽  
Ebba Nexø
Keyword(s):  
Malignant Melanoma ◽  
Melanoma Cell ◽  
Reverse Transcription ◽  
Peripheral Blood ◽  
Pcr Amplification ◽  
Melanoma Cells ◽  
Rt Pcr ◽  
Blood Samples ◽  
Reverse Transcription Pcr

Abstract Background: Reverse transcription-PCR (RT-PCR) amplification of melanoma cell-specific mRNA can detect melanoma cells in the peripheral blood of patients with malignant melanoma. We present a method to quantify mRNA coding for the melanoma-specific melanoma antigen recognized by T cells #1 (MART-1) in RNA isolated from peripheral blood. Methods: To establish a calibration curve, we measured the concentration of MART-1 mRNA in SK-MEL-28 melanoma cells grown in vitro by competitive RT-PCR. Serial dilutions of these cells were used as calibrators in the assay. The assay was conducted by adding a fixed amount of a RNA internal standard to RNA isolated from either peripheral blood or the calibrators before RT-PCR amplification with MART-1 primers in a nested PCR design. The amount of MART-1 mRNA in blood samples was calculated from the calibration curve. Results: Addition of melanoma cells grown in vitro to blood from healthy donors demonstrated that the method can detect a single SK-MEL-28 melanoma cell in 1 mL of blood (1.5 × 10−21 mol MART-1 mRNA/mL). MART-1 mRNA was observed in 4 of 12 blood samples from patients with malignant melanoma, at concentrations of 3–18 × 10−21 mol MART-1 mRNA/mL of blood. No MART-1 mRNA was detected in blood samples from 25 controls without malignant melanoma. Intra- and interassay CVs were 15% (n = 12; mean = 44 × 10−21 mol MART-1 mRNA/mL) and 33% (15 samples analyzed in two different analytical runs; mean = 30 × 10−21 mol MART-1 mRNA/mL), respectively. Conclusions: Our method is the first competitive RT-PCR assay for quantification of melanoma cells in blood samples that compensates for the variation of both the reverse transcription and PCR reactions. The method allows the inclusion of control samples for continuous quality assessment.

scholarly journals Molecular staging of prostatic cancer with RT-PCR assay for prostate-specific antigen in peripheral blood and lymph nodes: 5 Year follow-up

2007 ◽  
Vol 60 (10) ◽  
Author(s):  
Luis Martínez-Piñeiro ◽  
Montserrat Martínez-Gomariz ◽  
Emilio Rios ◽  
María L. Picazo ◽  
Hugo R. Arriaga ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Prostate Specific Antigen ◽  
Peripheral Blood ◽  
Prostatic Cancer ◽  
Specific Antigen ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Molecular Staging

The value of multi-marker reverse transcriptase-polymerase chain reaction (MM RT-PCR) assay in lymphatic drainage (LY) and peripheral blood (BL) for molecular staging of melanoma patients (pts) after lymph node dissection (LND)

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 8053-8053
Author(s):  
W. Ruka ◽  
Z. I. Nowecki ◽  
P. Rutkowski ◽  
J. Kulik ◽  
E. Lorenc ◽  
...  
Keyword(s):  
Lymph Node ◽  
Regional Lymph Node ◽  
Disease Free Survival ◽  
Melanoma Cells ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Melanoma Patients ◽  
Pcr Assays ◽  
Positive Results

8053 Background: We assessed the presence of melanoma cells in LY and BL in melanoma pts after LND with MM RT-PCR assay for the evaluation of disease outcomes. Methods: Between 05/2002 and 06/2005 we collected 24-hr LY in 207 stage III melanoma patients after radical LND (91 - completion LND after positive sentinel node biopsy - CLND and 116 - therapeutic LND for clinically/cytologically detected regional lymph node (LN) metastases - TLND). In 101 pts (since 02/2004) we have simultaneously collected a 5 ml sample of BL. In order to detect melanoma cells, RT-PCR assays with primers specific for tyrosinase, MART1 (MelanA) and uMAGE mRNA were applied. The drain fluid or BL sample was assumed to be positive if the presence of at least 1 marker was detected in the assay. Median follow-up time for survivors was 13 months (range: 4 - 39 months). Results: The LY MM RT-PCR assay results were positive in 57/207 pts (27.5%) - in 13 for tyrosinase only, in 17 for MART1 only, in 10 for uMAGE only, in 13 for a combination of the two markers and in 4 simultaneously for all three markers. We observed a significantly higher rate of recurrences in pts with positive LY MM RT-PCR results (44/57 cases; 77%) than in those with negative results (72/150 cases; 48%; p=0.0004). Positive results of the LY MM RT-PCR correlated with established predictive factors: the number of involved lymph nodes (p=0.03), extracapsular extension of LN metastases (p=0.004) and type of LND (CLND vs. TLND; p = 0.0004). We observed a significant relationship between positive LY MM RT-PCR results and shorter overall (OS) and disease free survival (DFS), both in univariate and multivariate analyses. In 107 pts, in whom the LY and BL MM RT-PCR assays were analyzed concurrently, we found positive results of the LY RT-PCR assay in 33 cases (32.7%) and the BL RT-PCR assay in 24 cases (23.7%) (17 of these positive results were detected in both tests), but longer follow-up time is necessary to perform a survival analysis in this group. Conclusions: We observed positive results of MM RT-PCR assay for melanoma cells in approximately 28% of LY after LND, which correlated significantly with early melanoma recurrences and shorter survival. No significant financial relationships to disclose.

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