Photoacoustic detection of circulating tumor cells in blood samples of stage III melanoma patients.

2016 ◽  
Vol 34 (15_suppl) ◽  
pp. 9555-9555
Author(s):  
John Andrew Viator ◽  
Benjamin Goldschmidt ◽  
Ahmad A. Tarhini ◽  
Cindy Sander ◽  
Martin Sanders
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Stage Iii ◽  
Photoacoustic Detection ◽  
Blood Samples ◽  
Melanoma Patients ◽  
Stage Iii Melanoma

scholarly journals Prospective Molecular Profiling of Circulating Tumor Cells from Patients with Melanoma Receiving Combinatorial Immunotherapy

2019 ◽  
Vol 66 (1) ◽  
pp. 169-177 ◽  
Author(s):  
Selena Y Lin ◽  
Shu-Ching Chang ◽  
Stella Lam ◽  
Romela Irene Ramos ◽  
Kevin Tran ◽  
...  
Keyword(s):  
Metastatic Melanoma ◽  
Tumor Cells ◽  
Clinical Outcomes ◽  
Circulating Tumor Cells ◽  
Progressive Disease ◽  
Molecular Profiling ◽  
Braf V600e ◽  
Blood Samples ◽  
Mrna Profiling ◽  
Melanoma Patients

Abstract BACKGROUND Blood molecular profiling of circulating tumor cells (CTCs) can enable monitoring of patients with metastatic melanoma during checkpoint inhibitor immunotherapy (CII) and in combination with targeted therapies. We developed a microfluidics-based CTC platform to explore CTC profiling utility in CII-treated patients with melanoma using a melanoma messenger RNA (mRNA)/DNA biomarker panel. METHODS Blood samples (n = 213) were collected prospectively from 75 American Joint Committee on Cancer-staged III/IV melanoma patients during CII treatment and those enriched for CTCs. CTC profiling was performed using 5 known melanoma mRNA biomarkers and BRAF V600E DNA mutation. CTC biomarker status associations with clinical outcomes were assessed. RESULTS CTCs were detected in 88% of blood samples from patients with melanoma. CTC-derived biomarkers and clinical variables analyzed using classification and regression tree analysis revealed that a combination of lactate dehydrogenase, CTC-mRNA biomarkers, and tumor BRAF–mutation status was indicative of clinical outcomes for patients with stage IV melanoma (n = 52). The panel stratified low-risk and high-risk patients, whereby the latter had poor disease-free (P = 0.03) and overall survival (P = 0.02). Incorporation of a DNA biomarker with mRNA profiling increased overall CTC-detection capability by 57% compared to mRNA profiling only. RNA sequencing of isolated CTCs identified significant catenin beta 1 (CTNNB1) overexpression (P <0.01) compared to nondisease donor blood. CTC-CTNNB1 was associated with progressive disease/stable disease compared to complete-responder patient status (P = 0.02). Serial CTC profiling identified subclinical disease in patients who developed progressive disease during treatment/follow-up. CONCLUSIONS CTC-derived mRNA/DNA biomarkers have utility for monitoring CII, targeted, and combinatorial therapies in metastatic melanoma patients.

Photoacoustic detection of circulating melanoma cells as a predictor of metastasis in stage III patients.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e21053-e21053
Author(s):  
John Andrew Viator ◽  
Martin Sanders ◽  
Ahmad A. Tarhini ◽  
Cindy Sander ◽  
Robert Hugh Edgar ◽  
...  
Keyword(s):  
Laser Light ◽  
Melanoma Cells ◽  
Ultrasonic Waves ◽  
Flow Cytometer ◽  
Disease State ◽  
Stage Iii ◽  
Blood Samples ◽  
Serial Blood ◽  
Melanoma Patients ◽  
Stage Iii Melanoma

e21053 Background: Circulating tumor cells have been correlated with disease state and distant metastatic spread in cancer patients. We postulated that enumerating circulating melanoma cells (CMCs) may predict the onset of distant metastasis in Stage III patients. We detected CMCs using our photoacoustic flow cytometer, in which we irradiated enriched blood samples with nanosecond pulsed laser light. While there is no effect on non-optically active leukocytes, absorption of laser light by pigmented melanoma cells resulted in robust ultrasonic waves that indicated CMCs in the sample. Methods: We tested 32 archived samples from 9 Stage III melanoma patients using our photoacoustic flow cytometer. Each patient had between 2 and 6 serial blood samples. We used a pulsed Nd:YAG laser to irradiate mononuclear cells in suspension and under flow. The number of CMCs detected after testing was recorded, indicating the time sequence of circulating tumor cell activity. Results: The numbers of CTCs for each sample is shown in the table below. The ultimate disease state, whether the patient became metastatic or not, was blinded to the investigators who performed the photoacoustic tests. One sample for patient 3 indicated 63 CMCs, though this test was known to be contaminated and had an unknown number of false detections. Conclusions: We found that patients who had a series of more than 4 CMCs were more likely to become metastatic than those patients who tested for 4 CMCs for fewer, indicating that a sequence of CMC detections in serial blood draws provides a potentially strong predictor of metastasis in Stage III melanoma patients warranting further investigation at this and lower stages of melanoma. We are developing a more rigorous model based on time series analysis of CMCs for prediction of metastasis. [Table: see text]

scholarly journals Double venipuncture is not required for adequate S-100B determination in melanoma patients

BioTechniques ◽  
2020 ◽  
Vol 69 (5) ◽  
pp. 371-378
Author(s):  
Samantha Damude ◽  
Anneke C Muller Kobold ◽  
Esther Bastiaannet ◽  
Schelto Kruijff ◽  
Harald J Hoekstra ◽  
...  
Keyword(s):  
Cancer Staging ◽  
Stage Iii ◽  
Tube System ◽  
Serum Samples ◽  
Blood Samples ◽  
Significant Difference ◽  
Melanoma Patients ◽  
The Mean ◽  
Stage Iii Melanoma

S-100B is used in melanoma follow-up. This serum biomarker is also present in adipocytes; therefore, subcutaneous adipocytes trapped in the needle before performing a venipuncture could contaminate the serum. The aim was to study the influence of adipocyte contamination on blood samples used for S-100B analysis, possibly resulting in falsely elevated S-100B values. A total of 294 serum samples were collected from 147 American Joint Committee on Cancer staging stage III melanoma patients. The mean difference between the first (dummy) and second tubes was 0.003 μg/l (p = 0.077), with a decrease in the second tube. Compared with the second tube, the S-100B level was higher in the first tube in 33.3% of the samples, equal in 36.8% of the samples and lower in 29.9% of the samples. No significant difference between the two consecutively drawn tubes was found. There seems to be no necessity of implementing a dummy tube system for accurate S-100B determination in melanoma patients.

Isolation and expansion of circulating tumor cells (CTC) from melanoma patients using a novel cell culture technique.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 10614-10614 ◽  
Author(s):  
John R. McGregor ◽  
Wolfram E. Samlowski ◽  
Shweta Tharkar ◽  
Sreekanth Donepudi ◽  
Soldano Ferrone
Keyword(s):  
Cell Culture ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Molecular Study ◽  
Immunofluorescence Assay ◽  
Buffy Coat ◽  
Culture Technique ◽  
Blood Samples ◽  
Patients Âgés ◽  
Melanoma Patients

10614 Background: Identification of rare (>2-5) circulating tumor cells (CTC) in 7.5 ml blood by immunofluorescence assay (IFA) correlates with a poor prognosis in colon, breast, prostate and lung cancer. Changes in CTC count during treatment also predict the eventual patient progression and survival in these cancers. Existing assays do not detect melanoma CTC, however. In addition, isolation of viable CTC remains problematic. To overcome these limitations we attempted to develop novel melanoma CTC assays, using IFA and cell culture approaches. Methods: Blood samples were obtained from patients and controls following informed consent. The buffy coat (white cells + tumor) was isolated by Ficoll/Hypaque centrifugation, and split into 6 replicate cultures in proprietary TrueCells medium. After 21 days in culture, tumor colonies were counted, and stained for melanoma and leukocyte markers. Buffy coat cells from parallel blood samples were stained with a panel of CSPG4-specific mAb (a pan-melanoma marker) on ultraclean glass slides for analysis by immunofluorescence microscopy. Results: Blood samples were obtained from 16 melanoma patients, ages 28-87. Eight patients were men and 8 were women. CSPG4+ events (>2) were detected in 8/16 patients by IFA (range 0-52). In contrast, tumor cell colonies of >50 cells grew in 12 out of 16 patients with Stage 3 or 4 melanoma (range 0-1054), shown in Table. Cells isolated from CTC colonies produced melanin, stained for CSPG4 and other melanoma markers, but not for leukocyte markers. Control cultures grew no tumor colonies. Conclusions: Our pilot study shows that melanoma CTC can be identified by both IFA and cultured from blood in many patients with stage 3 or 4 melanoma. These CTC exhibited cytologic characteristics diagnostic of melanoma. The culture assay may represent a useful means of enumerating, isolating, and expanding viable melanoma CTC for further molecular study. [Table: see text]

Detection of circulating melanoma cells by specific amplification of tyrosinase complementary DNA is not a reliable tumor marker in melanoma patients: a clinical two-center study.

1997 ◽  
Vol 15 (8) ◽  
pp. 2818-2825 ◽  
Author(s):  
R Gläser ◽  
K Rass ◽  
S Seiter ◽  
A Hauschild ◽  
E Christophers ◽  
...  
Keyword(s):  
Basal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cell Lines ◽  
Melanoma Cells ◽  
Rt Pcr ◽  
Blood Samples ◽  
Melanoma Patients ◽  
Center Study

PURPOSE Recently, the reverse-transcription polymerase chain reaction (RT-PCR) of tyrosinase messenger RNA (mRNA) was reported to be a useful tool for the detection of circulating tumor cells in the peripheral blood of melanoma patients. Our aim was to evaluate critically the diagnostic value of this marker by investigating a significant number of patients in different stages of the disease in a two-center study. PATIENTS AND METHODS Different techniques of blood collection, RNA isolation, and RT-PCR were compared, and the detectability of tyrosinase mRNA was tested using nine different melanoma cell lines. The sensitivity of the method was confirmed by blood spiking experiments and the specificity by restriction enzyme analysis. Subsequently, a total of 153 blood samples from 137 individuals (30 healthy subjects, five basal cell carcinoma, and 102 melanoma patients) were investigated. RESULTS The detection level of melanoma cells differed between the cell lines tested. However, we could reproducibly detect single melanoma cells by spiking whole blood samples from healthy volunteers. One of 43 patients with primary melanoma (2.3%), zero of 15 patients with regional metastasis (0%), and 12 of 44 patients with advanced disease (27.3%) were found to be RT-PCR positive. All blood samples obtained from controls and patients with basal cell carcinoma were tyrosinose mRNA negative. CONCLUSION Our data support the recent doubts that the detection of circulating tumor cells in melanoma patients using the tyrosinase mRNA RT-PCR is not sensitive enough to be used either as a melanoma progression marker in early stages of the disease or to monitor therapy in advanced stages of the disease.

scholarly journals Photoacoustic detection of circulating melanoma cells in late stage patients

2020 ◽  
Vol 13 (06) ◽  
pp. 2050023 ◽  
Author(s):  
John A. Viator ◽  
Marc Hazur ◽  
Andrea Sajewski ◽  
Ahmad Tarhini ◽  
Martin E. Sanders ◽  
...  
Keyword(s):  
Stage Iv ◽  
Melanoma Cells ◽  
The Body ◽  
Stage Iii ◽  
Photoacoustic Detection ◽  
Blood Samples ◽  
Secondary Tumors ◽  
Melanoma Patients ◽  
Rare Cells ◽  
Disease Free

Melanoma is the deadliest skin cancer and is responsible for over 7000 deaths in the US annually. The spread of cancer, or metastasis, is responsible for these deaths, as secondary tumors interrupt normal organ function. Circulating tumor cells, or those cells that spread throughout the body from the primary tumor, are thought to be responsible for metastasis. We developed an optical method, photoacoustic flow cytometry, in order to detect and enumerate circulating melanoma cells (CMCs) from blood samples of patients. We tested the blood of Stage IV melanoma patients to show the ability of the photoacoustic flow cytometer to detect these rare cells in blood. We then tested the system on archived blood samples from Stage III melanoma patients with known outcomes to determine if detection of CMCs can predict future metastasis. We detected between 0 and 66 CMCs in Stage IV patients. For the Stage III study, we found that of those samples with CMCs, two remained disease free and five developed metastasis. Of those without CMCs, six remained disease free and one developed metastasis. We believe that photoacoustic detection of CMCs provides valuable information for the prediction of metastasis and we postulate a system for more accurate prognosis.

PP 3 S-100B concentrations predict disease specific survival in AJCC Stage III melanoma patients

2011 ◽  
Vol 47 ◽  
pp. S21
Author(s):  
S. Kruijff ◽  
E. Bastiaannet ◽  
M. Speijers ◽  
I. Kema ◽  
R. van Ginkel ◽  
...  
Keyword(s):  
Stage Iii ◽  
Disease Specific Survival ◽  
Ajcc Stage ◽  
Melanoma Patients ◽  
Stage Iii Melanoma ◽  
Disease Specific

scholarly journals Circulating tumor cells: a valuable marker of poor prognosis for advanced nasopharyngeal carcinoma

2019 ◽  
Vol 25 (1) ◽  
Author(s):  
Guoping Ou ◽  
Shan Xing ◽  
Jianpei Li ◽  
Lin Zhang ◽  
Shulin Chen
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Roc Curve ◽  
Poor Prognosis ◽  
Regression Models ◽  
Prognostic Value ◽  
Stage Iv ◽  
Stage Iii ◽  
Ebv Dna

Abstract Purpose To evaluate the prognostic value of circulating tumor cells (CTCs) in nasopharyngeal carcinoma (NPC). Methods Cox’s proportional hazards regression models were used to identify whether CTCs was a poor prognostic factor for NPC. Chi-square tests were used to analyze and compare the distribution characteristics of CTCs in NPC. ROC curve was used to estimate the cut-off point of CTCs. Kaplan-Meier survival analyses were used to observe the prognostic value of CTCs alone and in combined with Epstein-Barr Virus DNA (EBV-DNA). Results CTCs was confirmed to be an independent risk factor for poor prognosis of NPC by Cox’s regression models that enrolled 370 NPC cases and took age, gender, EBV-DNA and CTCs as variables. The proportion of CTCs in stage IV NPC was statistically different from that in stage III; the cut-off point of CTCs between stage IV (288 cases) and stage III (70 cases) NPC estimated by ROC curve was 0.5. The prognosis of advanced NPC patients became worse with the increase of CTCs count. The combined detection of CTCs and EBV-DNA could better predict the prognosis of NPC compared with the single detection of EBV-DNA.

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