Relationship among circulating tumor cells,CEA and overall survival in patients with metastatic colorectal cancer

BackgroundWe previously reported results of a prospective trial evaluating the significance of circulating tumor cells (CTCs) in patients with metastatic colorectal cancer (mCRC). This secondary analysis assessed the relationship of the CTC number with carcinoembryonic antigen (CEA) and overall survival.Patients and methodsPatients with mCRC had CTCs measured at baseline and specific time points after the initiation of new therapy. Patients with a baseline CEA value ≥10 ng/ml and CEA measurements within ±30 days of the CTC collection were included.ResultsWe included 217 patients with mCRC who had a CEA value of ≥10 ng/ml. Increased baseline CEA was associated with shorter survival (15.8 versus 20.7 months, P = 0.012). Among all patients with a baseline CEA value of ≥25 ng/ml, patients with low baseline CTCs (<3, n = 99) had longer survival than those with high CTCs (≥3, n = 58; 20.8 versus 11.7 months, P = 0.001). CTCs added prognostic information at the 3–5- and 6–12-week time points regardless of CEA. In a multivariate analysis, CTCs at baseline but not CEA independently predicted survival and both CTCs and CEA independently predicted survival at 6–12 weeks.ConclusionsThis study demonstrates that both CEA and CTCs contribute prognostic information for patients with mCRC.     

Clinical value of circulating endothelial cells and circulating tumor cells in metastatic breast cancer patients treated first line with bevacizumab and chemotherapy

BackgroundWe investigated whether circulating tumor cells (CTCs) and circulating endothelial cells (CECs) predict clinical outcome of first-line chemotherapy combined with bevacizumab in metastatic breast cancer patients.Patients and methodsIn a French substudy of the MO19391 trial, CTC and CEC counts (CellSearch system) at baseline and changes after two cycles of treatment were correlated with time to progression (TtP).ResultsCTC and CEC levels were not correlated in the 67 patients included. At baseline, CTC positivity was a significant prognostic marker for TtP at a threshold of 3 CTC/7.5 ml (P < 0.05) but not at 5 CTC/7.5 ml (P = 0.09). Baseline CEC levels (median 17 CEC/4 ml, range 1–769) were associated with age ≥45 years (P = 0.01), elevated lactate dehydrogenase (P < 0.01) and not with TtP at any threshold. Changes of CTC count during treatment were not a surrogate of TtP, with any of the model tested (threshold based or relative decrease in percent). However, increase in CEC count was associated with improved TtP, at the threshold of 20 CEC/4 ml (P < 0.01).ConclusionBevacizumab combined with first-line chemotherapy may modify the predictive value of CTC during treatment possibly due to impaired tumor cells intravasation through vessels endothelium. Variations in CEC levels appear to be a promising early surrogate marker of TtP under antiangiogenic treatment.     

Determination of cut-offs for circulating tumor cell measurement in metastatic cancer

The metastatic transformation of epithelial tumors progresses through various steps leading to the generation of circulating tumor cells (CTCs). Measurement of CTCs in the peripheral blood is being increasingly recognized as a promising tool in breast oncology. Several studies have evaluated the prognostic significance of CTCs in newly diagnosed metastatic breast cancer (MBC) patients. The IC 2006-04 was a high-powered, prospective, multicenter, observational study conceived to assess CTC changes in women with MBC treated with first-line chemotherapy. Levels ≥ 5 CTCs/7.5 ml blood at baseline and before the second cycle of treatment were independent prognostic factors associated with shorter progression-free and overall survival. This study provides further level II evidence for the clinical and prognostic value of CTCs in MBC, confirming data from earlier small studies. It also provides proof that CTCs should be investigated in ongoing interventional trials to see if better patient outcomes can be attained by altering treatment based on CTC levels.     

Determination of cut-offs for circulating tumor cell measurement in metastatic cancer

Evaluation of: Pierga JY, Hajage D, Bachelot T et al. High independent prognostic and predictive value of circulating tumor cells compared with serum tumor markers in a large prospective trial in first-line chemotherapy for metastatic breast cancer patients. Ann. Oncol. DOI: 10.1093/annonc/mdr263 (2011) (Epub ahead of print).

The metastatic transformation of epithelial tumors progresses through various steps leading to the generation of circulating tumor cells (CTCs). Measurement of CTCs in the peripheral blood is being increasingly recognized as a promising tool in breast oncology. Several studies have evaluated the prognostic significance of CTCs in newly diagnosed metastatic breast cancer (MBC) patients. The IC 2006-04 was a high-powered, prospective, multicenter, observational study conceived to assess CTC changes in women with MBC treated with first-line chemotherapy. Levels ≥5 CTCs/7.5 ml blood at baseline and before the second cycle of treatment were independent prognostic factors associated with shorter progression-free and overall survival. This study provides further level II evidence for the clinical and prognostic value of CTCs in MBC, confirming data from earlier small studies. It also provides proof that CTCs should be investigated in ongoing interventional trials to see if better patient outcomes can be attained by altering treatment based on CTC levels.     

High independent prognostic and predictive value of circulating tumor cells compared with serum tumor markers in a large prospective trial in first-line chemotherapy for metastatic breast cancer patients

BackgroundCirculating tumor cells (CTCs) are a prognostic marker in metastatic breast cancer, but comparisons with serum tumor markers (CA 15-3, carcinoembryonic antigen and lactate dehydrogenase) variations are needed.Patients and methodsCTCs were counted with CellSearch® at baseline, before cycle 2 (C2) and cycle 3 or 4 (C3/4) in 267 metastatic breast cancer patients on first-line chemotherapy with/without targeted therapy.ResultsBaseline CTC detection rate was 65% with ≥1 CTC/7.5 ml threshold and 44% with ≥5 CTC/7.5 ml and was independent of subtypes (luminal, triple negative, human epithelial growth factor receptor 2 (HER2)+). CTCs were associated with tumor markers, bone/liver involvement, tumor burden and performance status. CTC detection ≥1 CTC/7.5 ml was a strong prognostic factor for progression-free survival (PFS), P < 0.0001. Threshold of CTC ≥5 was statistically significant for PFS and overall survival (OS), P = 0.03 on multivariate analysis. Among patients with ≥5 CTC/7.5 ml at baseline, 50% had <5 CTC/7.5 ml at C2. Changes were correlated with both PFS and OS (P < 0.0001). All patients receiving anti-HER2 therapy had <5 CTC/7.5 ml after three cycles of treatment.ConclusionThis is the largest prospective series validating the prognostic value of CTC independently from serum tumor marker. Elevated CTCs before C2 are an early predictive marker of poor PFS and OS, which could be used to monitor treatment benefit. CTC decrease under treatment seems stronger with targeted therapy.     

Apoptotic circulating tumor cells (CTCs) in the peripheral blood of metastatic colorectal cancer patients are associated with liver metastasis but not CTCs

Enumeration of circulating tumor cells (CTCs) by the CellSearch system provides prognostic information in metastatic colorectal cancer, regardless of metastatic site. We found that CTCs generally represent <1% of observed events with CellSearch analysis and adapted scoring criteria to classify other peripheral blood events. Examination of twenty two metastatic colorectal cancer patients'' blood revealed that patients with high CEA or liver metastases, but not lung or distant lymph node metastases, possessed significant numbers of apoptotic CTCs prior to treatment initiation by Fischer''s exact test. Six out of eleven patients with liver metastasis possessed apoptotic CTCs whereas one of nine patients with other metastases had measurable apoptotic CTCs. An elevated CTC number was not necessarily associated with apoptotic CTCs or CTC debris by Spearman''s correlation, suggesting the metastatic site rather than CTCs per se as contributing to the origin of these events.     

Detection of circulating tumor cells in patients with urothelial cancer

Background: Approximately 50% of patients with metastatic urothelialcancer (UC) respond to chemotherapy and several months of therapyis required to assess for radiographic response. Blood-basedbiomarkers may identify patients in whom a specific therapyprovides clinical benefit, and this study sought to characterizecirculating tumor cells (CTCs) in patients with metastatic UC. Patients and methods: Peripheral blood from patients with metastaticUC was evaluated for CTCs using the CellSearch system. We assessedfor associations between CTC counts and the number and sitesof metastatic disease. Results: CTC evaluations were carried out in 33 patients withmetastatic UC. Fourteen of 33 patients (44%; 95% confidenceinterval 27% to 59%) had a positive assay (range 0–87cells/7.5 ml of blood) with 10 patients (31%) having five ormore CTCs. A significantly higher number of CTCs was seen inpatients with two or more sites of metastases compared withthose with less than one or one site of metastases (3.5 versus0, P = 0.04). Conclusions: CTCs, detected by antibody capture technology,are present in 44% of patients with metastatic UC. Higher numbersof CTCs are seen in patients with a greater number of metastaticsites. One-third of patients have five or more CTCs providinga potential early marker to monitor response to chemotherapy. Key words: bladder cancer, circulating tumor cells, urothelial cancer Received for publication May 23, 2008. Revision received July 10, 2008. Accepted for publication August 7, 2008.     

Circulating tumour cell (CTC) counts as intermediate end points in castration-resistant prostate cancer (CRPC): a single-centre experience

Background: The purpose of this study was to evaluate the associationof circulating tumour cell (CTC) counts, before and after commencingtreatment, with overall survival (OS) in patients with castration-resistantprostate cancer (CRPC). Experimental design: A 7.5 ml of blood was collected beforeand after treatment in 119 patients with CRPC. CTCs were enumeratedusing the CellSearch®System. Results: Higher CTC counts associated with baseline characteristicsportending aggressive disease. Multivariate analyses indicatedthat a CTC     

Circulating Tumor Cells Following First Chemotherapy Cycle: An Early and Strong Predictor of Outcome in Patients With Metastatic Breast Cancer

We investigated the prognostic significance of circulating tumor cells (CTCs) determined immediately before the second cycle of chemotherapy in patients with metastatic breast cancer (MBC). The CTC counts were taken at baseline, before the first cycle of chemotherapy (CTC‐0), and on day 21 before commencing the second cycle of chemotherapy (CTC‐21) in consecutive MBC patients. The study's primary objectives were to analyze relationships between CTC‐21 count and overall survival (OS). Based on the current literature, the CTC measurements were dichotomized as 0–4 versus ≥5 CTCs. Of 117 patients recruited, 99 were evaluable. Patients with 0–4 CTCs on day 21 had a significantly better OS than those with ≥5 CTCs (median OS: 38.5 months vs. 8.7 months). They also had a significantly better progression‐free survival (PFS; median: 9.4 months vs. 3.0 months) and clinical benefit rate (77% vs. 44%). The OS of patients whose baseline CTCs were ≥5 but dropped to <5 on day 21 was apparently similar to those who had <5 CTCs at baseline. In a Cox regression analysis, CTC‐21 was the only independent variable significantly predicting OS and PFS. Our data indicate that CTCs determined immediately before the second cycle of chemotherapy is an early and strong predictor of treatment outcome in MBC patients.     

外周血循环肿瘤细胞在晚期肺癌患者的预测及预后价值研究

目的应用免疫磁珠阴性富集技术结合荧光细胞化学染色方法,检测肺癌患者外周静脉血中循环肿瘤细胞（CTC）的敏感性及特异性;分析肺癌患者CTC水平与临床疗效的相关性。方法2007年7月～2008年4月在北京协和医院呼吸内科肺癌中心诊断的144例肺癌患者作为研究对象,并设健康对照组35例、良性肺疾病组28例。采集上述各组静脉血标本,应用免疫磁珠阴性富集分离及免疫荧光细胞化学染色方法检测静脉血中CK18＋／CD45-／CTC计数,凡≥1判定为阳性。随访上述52例入院化疗患者治疗后CTC,与其临床疗效、生存进行相关性分析。结果健康和良性肺疾病组CTC检测阳性分别为2例（5．71％）及1例（3．57％）,肺癌组未接受抗肿瘤治疗前CTC的阳性率64．5％（93／144）。其中ⅢA、ⅢB及Ⅳ期分别为32．0％、56．8％及78．0％。腺癌、鳞癌、小细胞癌CTC阳性率分别为66．7％、70％及62％。52例肺癌患者进行每周期化疗后CTC检测,疗前CTC阳性率69．2％（36／52）,3个周期化疗后下降至6．5％（3／46）,显示CTC与疗效相关。结论本检测方法简便,敏感性为93．1％,特异性为72．8％;CTC阳性率与疾病分期、吸烟、疗效及转移相关,CTC水平越高可能提示预后不良。     

Thymidylate synthase expression in circulating tumor cells: A new tool to predict 5‐fluorouracil resistance in metastatic colorectal cancer patients

Thymidylate synthase (TYMS) is an important enzyme for 5‐fluorouracil (5‐FU) metabolism in metastatic colorectal cancer (mCRC) patients. The search for this enzyme in circulating tumor cells (CTCs) can be a powerful tool to follow‐up cancer patients. mCRC patients were enrolled before the beginning of 5‐FU‐based chemotherapy. The blood was filtered on Isolation by Size of Epithelial Tumor Cells (ISET), and the analysis of TYMS expression in CTCs was made by immunocytochemistry. Additionally, we verified TYMS staining in primary tumors and metastases from the same patients. There were included 54 mCRC patients and 47 of them received 5‐FU‐based chemotherapy. The median CTCs number was 2 per mL. We were not able to analyze immunocytochemistry in 13 samples (9 patients with absence of CTCs and 4 samples due to technical reasons). Therefore, TYMS expression on CTCs was analyzed in 34 samples and was found positive in 9 (26.5%). Six of these patients had tumor progression after treatment with 5‐FU. We found an association between CTC TYMS staining and disease progression (DP), although without statistical significance (P = 0.07). TYMS staining in primary tumors and metastases tissues did not have any correlation with disease progression (P = 0.67 and P = 0.42 respectively). Patients who had CTC count above the median (2 CTCs/mL) showed more TYMS expression (P = 0.02) correlating with worse prognosis. Our results searching for TYMS staining in CTCs, primary tumors and metastases suggest that the analysis of TYMS can be useful tool as a 5‐FU resistance predictor biomarker if analyzed in CTCs from mCRC patients.     

Circulating tumor cells in high-risk nonmetastatic colorectal cancer

The identification of patients at higher risk of recurrence after primary colorectal cancer resection is currently one of the challenges facing medical oncologists. Circulating tumor cell (CTC) may represent a surrogate marker of an early spread of disease in patients without overt metastases. Thirty-seven high-risk stages II–III colorectal cancer patients were evaluated for the presence of CTC. Enumeration of CTCs in 7.5 ml of blood was carried out with the FDA-cleared CellSearch system. CTC count was performed after primary tumor resection and before the start of adjuvant therapy. CTC was detected in 22 % of patients with a significant correlation with regional lymph nodes involvement and stage of disease. No significant correlation was found among the presence of CTC and other clinicopathological parameters. These data suggest that CTCs detection might help in the selection of high-risk stage II colorectal cancer patient candidates for adjuvant chemotherapy.     

Evaluation of circulating tumor cells in patients with breast cancer: multi-institutional clinical trial in Japan

BACKGROUND: With the development of the CellSearch System, it has become possible to measure circulating tumor cell (CTC) levels with high reproducibility, and the CTC test is currently being used clinically for patients with metastatic breast cancer in the United States. It is imperative that the clinical significance of the CTC test also be examined in Japan. METHODS: Using the CellSearch System, CTC levels were evaluated in 57 healthy individuals and patients with benign breast disease; 30 patients with primary breast cancer (stages 1-3); and 38 patients with metastatic breast cancer. First, the relationship between CTC levels and the presence of metastasis was examined using a cutoff score of 2 CTCs per 7.5 ml whole blood. Then, the patients with metastatic breast cancer were divided into two groups, using a cutoff score of 5 CTCs per 7.5 ml blood, and progression-free survival (PFS) and overall survival (OS) were compared in the two groups. RESULTS: When the clinical cutoff score was set at 2 CTCs per 7.5 ml blood, 0% of the healthy individuals and patients with benign breast disease (0/57), 3.3% of the patients with primary breast cancer (1/30), and 50% of the patients with metastatic breast cancer (19/38) were identified as as having 2 CTCs per 7.5 ml blood. Additionally, with a cutoff score of 5 CTCs, 11 patients were reported to have 5 or more CTCs and both PFS (P = 0.0036) and OS (P = 0.04) were worse for this patient population than for the population with fewer than 5 CTCs. CONCLUSION: As concluded in a similar clinical trial in the United States, for patients with breast cancer, measuring CTC levels can be both an accurate indicator of metastases and an important measure of patient prognosis.     

Parallel Evaluation of Circulating Tumor DNA and Circulating Tumor Cells in Metastatic Colorectal Cancer

Background

Tissue biopsy is the gold standard for tumor genotyping, but it is an invasive procedure providing a single snapshot into tumor heterogeneity. Liquid biopsy approaches, encompassing the analysis of circulating tumor DNA (ctDNA) or circulating tumor cells (CTCs), have been proposed as an alternative, with the potential of providing a comprehensive portrait of the tumor molecular landscape. In metastatic colorectal cancer (mCRC), both CTCs and ctDNA analysis have been investigated, but comparative analyses are limited.

Denosumab treatment is associated with the absence of circulating tumor cells in patients with breast cancer

Background

The presence of circulating tumor cells (CTCs) in patients with breast cancer correlates to a bad prognosis. Yet, CTCs are detectable in only a minority of patients with progressive breast cancer, and factors that influence the abundance of CTCs remain elusive.

Methods

We conducted CTC isolation and enumeration in a selected group of 73 consecutive patients characterized by progressive invasive breast cancer, high tumor load and treatment discontinuation at the time of CTC isolation. CTCs were quantified with the Parsortix microfluidic device. Clinicopathological variables, blood counts at the time of CTC isolation and detailed treatment history prior to blood sampling were evaluated for each patient.

Results

Among 73 patients, we detected at least one CTC per 7.5 ml of blood in 34 (46%). Of these, 22 (65%) had single CTCs only, whereas 12 (35%) featured both single CTCs and CTC clusters. Treatment with the monoclonal antibody denosumab correlated with the absence of CTCs, both when considering all patients and when considering only those with bone metastasis. We also found that low red blood cell count was associated with the presence of CTCs, whereas high CA 15-3 tumor marker, high mean corpuscular volume, high white blood cell count and high mean platelet volume associated specifically with CTC clusters.

Conclusions

In addition to blood count correlatives to single and clustered CTCs, we found that denosumab treatment associates with most patients lacking CTCs from their peripheral circulation. Prospective studies will be needed to validate the involvement of denosumab in the prevention of CTC generation.

     

循环肿瘤细胞在转移性乳腺癌中的表达及其与多种血清肿瘤标志物的关系

目的 探讨转移性乳腺癌的循环肿瘤细胞(CTC)和血清肿瘤标志物(CEA、CA153和CA125)的表达水平以及两者之间的相关性。方法 采用CellSearch自动检测系统检测93例转移性乳腺癌患者开始新治疗前的循环肿瘤细胞(CTC),同时采用电化学发光法检测这些患者血清肿瘤标志物(CEA、CA153和CA125)的表达水平。结果 CTC阳性率为60%(56/93),与患者激素受体的状态、Her-2的状态、既往治疗的线数以及内脏转移无关。CEA的阳性率为56%(52/93),ER或PR阳性的患者CEA的阳性率更高(χ²=4.550,P=0.045),与Her-2的状态、既往治疗的线数以及内脏转移无关;CA153的阳性率为47%(44/93),与患者激素受体的状态、Her-2的状态、既往治疗的线数以及内脏转移无关;CA125的阳性率为41%(38/93);既往治疗大于等于二线的患者CA125的阳性率更高(χ²=4.501,P=0.038),与患者激素受体的状态、Her-2的状态以及是否有内脏转移无关。CTC和CEA之间呈正相关(r=0.296,P=0.004);CTC和CA153之间呈正相关(r=0.286,P=0.005);CTCs和CA125两者之间无相关性(r=0.184,P=0.077)。结论 转移性乳腺癌患者的循环肿瘤细胞(CTC)检出率高,而且与血清肿瘤标志物(CEA和CA153)有明显相关性,提示联合检测转移性乳腺癌的循环肿瘤细胞(CTC)和血清肿瘤标志物(CEA和CA153)可能会对其治疗决策有所帮助,同时CTC有可能成为转移性乳腺癌新的治疗靶点。     

负性筛选策略与食管鳞癌循环肿瘤细胞检测

目的建立基于细胞学水平的循环肿瘤细胞负性筛选方法,检测食管鳞癌患者外周血循环肿瘤细胞。方法选用负性筛选策略,分步去除7．5ml食管鳞癌外周血样本中的红、白细胞,富集上皮来源的稀有细胞。比较3种免疫磁珠（Miltenyi、AVIVA和Xinzhi磁珠）去除白细胞、富集外周血循环肿瘤细胞的特点和效率。用经典的细胞化学染色鉴定循环肿瘤细胞。结果Miltenyi和AVIVA磁珠去除白细胞、富集肿瘤细胞效果较好。从富集后的食管鳞癌患者外周血稀有细胞中鉴定到形态各异的循环肿瘤细胞。结论负性筛选策略可以有效分离鉴定食管鳞癌患者外周血形态各异的循环肿瘤细胞。     

循环肿瘤细胞数目与血清中Cyfra21-1、CEA、Fg表达水平的关系

目的:探讨肿瘤患者外周血循环肿瘤细胞(CTCs)与常用的肿瘤指标细胞角质素19可溶片段（Cyfra21-1）、癌胚抗原（CEA）及凝血功能指标纤维蛋白原（Fg）表达水平的关系,比较CTCs阳性组和阴性组转移灶数目差异。方法:采集66例进展期晚期肺癌住院患者治疗前7.5ml外周血,保存于4℃冰箱中,24小时内由益善生物技术公司使用“免疫去除结合纳米过滤法”行CTCs检测。同时检测血清中Cyfra21-1、CEA、Fg水平,收集并分析CTCs数目与转移灶数目、临床T分期、转移淋巴结的关系。结果:纳入患者中CTCs检出率为86.4%。CTCs数目与Cyfra21-1水平有相关关系,相关系数为0.365（P=0.003）;CTCs数目与Fg水平相关,相关系数为0.330（P=0.007）;CEA与CTCs两变量无明显相关性。两组患者的肿瘤转移器官数目大于3个的比例有显著差异。结论:两组患者Cyfra21-1异常增高率有显著差异,且CTCs数目与Cyfra21-1表达水平正相关,可由Cyfra21-1初步估计CTCs水平。而外周血中Fg异常增高率无显著差异,但二者表达水平呈明显正相关,因此分析CTCs数目需要考虑Fg表达水平的影响。