Circulating Endothelial Cells Level Research Articles

Evaluation of Circulating Endothelial Cells as Direct Marker of Endothelial Damage in Allo-Transplant Recipients at High Risk of Hepatic Veno-Occlusive Disease/Sinusoidal Obstruction Syndrome

Sinusoidal obstruction syndrome (SOS), also known as veno-occlusive disease (VOD), is a rare but potentially fatal complication following allogenic hematopoietic cell transplantation (allo-HCT). Timely identification of SOS/VOD to allow for prompt treatment is critical, but identifying a VOD-predictive biomarker remains challenging. Given the pivotal role of endothelial dysfunction in SOS/VOD pathophysiology, the CECinVOD study prospectively evaluated levels of circulating endothelial cells (CECs) in patients undergoing allo-HCT with a myeloablative conditioning (MAC) regimen to investigate the potential of CEC level in predicting and diagnosing SOS/VOD. A total of 150 patients from 11 Italian bone marrow transplantation units were enrolled. All participants were age >18 years and received a MAC regimen, putting them at elevated risk of developing SOS/VOD. Overall, 6 cases of SOS/VOD (4%) were recorded. CECs were detected using the Food and Drug Administration-approved CellSearch system, an immunomagnetic selection-based platform incorporating ferrofluid nanoparticles and fluorescent-labeled antibodies, and were defined as CD146+, CD105+, DAPI+, or CD45-. Blood samples were collected at the following time points: before (T0) and at the end of conditioning treatment (T1), at neutrophil engraftment (T2), and at 7 to 10 days postengraftment (T3). For patients who developed VOD, additional samples were collected at any suspected or proven VOD onset (T4) and weekly during defibrotide treatment (T5 to T8). A baseline CEC count >17/mL was associated with an elevated risk of SOS/VOD (P = .04), along with bilirubin level >1.5 mg/mL and a haploidentical donor hematopoietic stem cell source. Postconditioning regimen (T1) CEC levels were elevated (P = .02), and levels were further increased at engraftment (P < .0001). Additionally, patients developing SOS/VOD after engraftment, especially those with late-onset SOS/VOD, showed a markedly higher relative increase (>150%) in CEC count. Multivariate analysis supported these findings, along with a high Endothelial Activation and Stress Index (EASIX) score at engraftment (T2). Finally, CEC kinetics corresponded with defibrotide treatment. After the start of therapy (T4), CEC levels showed an initial increase in the first week (T5), followed by a progressive decrease during VOD treatment (T6 and T7) and a return to pre-SOS/VOD onset levels at resolution of the complication. This prospective multicenter study reveals a low incidence of SOS/VOD in high-risk patients compared to historical data, in line with recent reports. The results from the CECinVOD study collectively confirm the endothelial injury in allo-HCT and its role in in the development of SOS/VOD, suggesting that CEC level can be a valuable biomarker for diagnosing SOS/VOD and identifying patients at greater risk of this complication, especially late-onset SOS/VOD. Furthermore, CEC kinetics may support treatment strategies by providing insight into the optimal timing for discontinuing defibrotide treatment.

Evaluation of Circulating Endothelial Cells (CECs) As Marker of Endothelial Damage in Allo-Transplanted Patients at High Risk of Hepatic Veno-Occlusive Disease/Sinusoidal Obstruction Syndrome (VOD/SOS): The Cecinvod Study

INTRODUCTION Sinusoidal obstruction syndrome (SOS), also known as veno-occlusive disease (VOD), is a potentially fatal complication after allogeneic stem cell transplantation (alloSCT). Identifying a predictive biomarker for VOD has been challenging. Since endothelial injury is considered one of the main pathogenic factors for VOD onset, with the CECinVOD prospective study we aimed to evaluate Circulating Endothelial Cells (CEC) in allo-transplanted patients (pts) at higher risk to develop VOD. METHODS From October 2020 to November 2022, 150 pts have been enrolled in the CECinVOD study from 11 Italian Bone Marrow Transplantation Units. All pts must be older than 18 years and undergoing myeloablative alloSCT. CECs were detected using the CellSearch system, the FDA-approved immunomagnetic selection approach incorporating ferrofluid nanoparticles (anti CD146) and fluorophore-labelled antibodies (anti CD105, CD45 and DAPI). CEC were defined as CD146+, CD105+, DAPI+ and CD45-. CEC were collected at the following timepoints: before conditioning regimen (T0), at the end of conditioning regimen and before alloSCT (T1), at the time of neutrophils engraftment (T2), and 7-10 days after engraftment (T3). In pts who developed VOD, additional timepoints were collected as follows: at any time of suspected or proven VOD onset (T4), and then weekly during Defibrotide treatment (T5-T8). SOS/VOD was defined according to the 2016 European Group for Blood and Marrow Transplantation criteria. RESULTS Pts' main characteristics are summarized in Table 1. Six out of 150 pts (4%) developed VOD during the follow up (4 “severe”, and 2 “very severe”). All pts were treated with Defibrotide, obtaining a complete remission in 5 of them, while 1 pt died due to VOD complications. Pts receiving TBI-based regimen were more likely to develop VOD compared to those receiving Treosulfan (10 to 14 g/m2) or Busulfan ev (9.6 to 12.8 mg/kg) (p 0.08). Similarly, higher baseline levels of bilirubin were associated with a higher incidence of VOD (p 0.08). Considering the CECs analysis, 615 samples were evaluated. At the enrollment, CECs levels were not related with any of clinical characteristics analyzed, except for the number of previous treatments. Indeed, those pts with 2 or more previous lines of treatments had higher levels of CECs (OR=0.53; p&amp;lt;0.001). Considering the different timepoints, conditioning regimen and alloSCT result in higher levels of CECs. Thus, CEC were higher at T1 than at T0 (p 0.02), as well as they were even higher at T2 than at T1 (p&amp;lt;0.0001) (Fig.1). Conversely, no significant differences have been observed between T3 and T2. No other clinical characteristic was correlated with CEC counts at the different timepoints. Pts who developed VOD had higher median levels of CEC at all the timepoints analyzed, but this difference has not achieved statistical significance, probably due to the low number of pts in the VOD group. At VOD onset, the pts always had an increase in CEC levels compared with the previous timepoint. After defibrotide treatment, the CEC levels increased in the first week, while they progressively decreased during the VOD treatment (T6 and T7, -50,7% and -71,5%, respectively). Recently, another endothelial activation marker has been considered: the Easix score. We didn't find any relationship between the Easix score at transplant and the VOD onset, even if a trend may be observed. We also investigated whether the CEC levels may be related with Easix score, but we didn't find any relationship at each timepoint. Interestingly, in a subgroup of pts CECs have been observed as a cluster of multiple cells. This data is firstly described in allo-SCT. Its functional significate remains unclear, but we found a relationship between CECs cluster and the number of CECs (p &amp;lt; 0.001). CONCLUSIONS The incidence of VOD in our prospective study was low with respect to the one reported in the literature. This can be explained by the changes in alloSCT from the past (better selection of the pts, lower use of TBI, higher use of reduced intensity regimen) and the retrospective nature of most of the previous reports. We show that CECs can be considered reliable marker of endothelial damage in alloSCT pts, highlighting the impact of previous treatments, the conditioning regimen, and allo-SCT itself. Increased CEC level may be helpful to confirm VOD diagnosis, as well as their monitoring may be useful to evaluate the response to the treatment for VOD.

Circulating Endothelial Cells are Associated with Thromboembolic Events in Patients with Antiphospholipid Antibodies.

Endothelial damage has been described in antiphospholipid antibody (aPL)-positive patients. However, it is uncertain whether circulating endothelial cells (CECs)-which are released when endothelial injury occurs-can be a marker of patients at high risk for thrombosis. Ninety-seven patients with aPL and/or systemic lupus erythematosus (SLE) were included. CECs were determined by an automated CellSearch system. We also assayed plasma levels of tissue factor-bearing extracellular vesicles (TF+/EVs) and soluble triggering receptor expressed on myeloid cells 1 (sTREM-1) as markers of endothelial dysfunction/damage. Patients' mean age was 46.1 ± 13.9 years, 77 were women. Thirty-seven had SLE and 75 patients were suffering from antiphospholipid syndrome. Thirty-seven percent of patients presented a medical history of arterial thrombosis and 46% a history of venous thromboembolism (VTE). Thirteen patients had increased levels of CECs (>20/mL), with a mean CEC level of 48.3 ± 21.3 per mL. In univariate analysis, patients with obesity or medical history of myocardial infarction (MI), VTE, or nephropathy had a significant increased CEC level. In multivariate analysis, obesity (odds ratio [OR] = 6.07, 95% confidence interval [CI]: 1.42-25.94), VTE (OR = 7.59 [95% CI: 1.38-41.66]), and MI (OR = 5.5 [95% CI: 1.1-26.6)] were independently and significantly associated with elevated CECs. We also identified significant correlations between CECs and other markers of endothelial dysfunction: sTREM-1 and TF+/EVs. This study demonstrated that endothelial injury assessed by the levels of CECs was associated with thromboembolic events in patients with aPL and/or autoimmune diseases.

Нарушения эндотелий-зависимой вазодилатации у больных с микрососудистой стенокардией

Background: disorders of both endothelium-dependent and endothelium-independent vasodilation play a role in the pathogenesis of microvascular angina (MVA), the contribution of which may be different. Aim: to study the processes of endothelium-dependent vasodilation (EDV) in patients with primary MVA. Patients and Methods: an open prospective study included 60 patients (mean age 57.3±6.4 years) with MVA, confirmed clinical picture (chest pain), coronary angiography data (the absence of coronary artery stenosis), positive stress test (pain syndrome and/or ST segment depression by ≥2 mm by treadmill exercise stress test), positron emission tomography (PET) myocardial perfusion imaging (MPI) with tests (cold pressor test and with adenosine). Patients with diseases that can lead to secondary microvascular dysfunction were not included in the study. To evaluate the processes of EDV, all patients underwent the following evaluations: peripheral arterial tonometry (PAT), assessment of circulating endothelial cells (CEC), high-sensitivity C-reactive protein (hsCRP), endothelin-1 (ET-1), total antioxidant status. Adding that, the results of a cold pressor test during PET were analyzed. Results: the majority of the examined patients were women — 81.7%. Most of the female patients included in the study (93.9%) were postmenopausal. In all patients with MVA, according to the data of PET MPI with cold pressor test, signs of EDV disorder were detected as a decrease in coronary blood flow through the three coronary arteries. During PET, the index of reactive hyperemia was reduced (&lt;1,67) in all examined patients and amounted to 1.43±0.15. According to the results of laboratory studies, there was an increase in the level of ET-1 and CEC — 3,335 [1,545; 3,952] fmol/L and 14±8 cells/3×105 leukocytes, respectively. The CRP level was 4,44 [1.02; 4.45] mg/L. The total antioxidant capacity was reduced in 53 (88%) patients, the average values of the indicator were 289,03±52,14 μmol/L. Conclusion: according to laboratory and instrumental studies, signs of endothelial dysfunction, namely disorders of EDV, were revealed in all examined patients with primary MVA. KEYWORDS: microvascular angina, peripheral arterial tonometry, endothelium-dependent vasodilation, positron emission tomography. FOR CITATION: Leonova I.A., Zakharova O.V., Boldueva S.A. Disorders of endothelium-dependent vasodilation in patients with microvascular angina. Russian Medical Inquiry. 2022;6(8):427–432 (in Russ.). DOI: 10.32364/2587-6821-2022-6-8-427-432.

Correlation between circulating endothelial cell level and acute respiratory distress syndrome in postoperative patients.

BACKGROUNDAcute respiratory distress syndrome (ARDS) is injury of alveolar epithelial cells and capillary endothelial cells caused by various factors, including endogenous and exogenous lung factors, leading to diffuse pulmonary interstitial and alveolar edema, and acute respiratory failure. ARDS involves alveolar epithelial cells and pulmonary interstitial capillary endothelial cells. Circulating endothelial cells (CECs) are the only marker that directly reflects vascular endothelial injury in vivo. There have been few studies on the correlation between peripheral blood CECs and ARDS at home and abroad. The lungs are the organs with the highest capillary density and the most endothelial cells, thus, it is speculated that when ARDS occurs, CECs are stimulated and damaged, and released into the circulatory system. AIMTo explore the correlation between CEC level and severity of ARDS in patients postoperatively.METHODSBlood samples were collected from all patients on day 2 (d2) and day 5 (d5) after surgery. The control group comprised 32 healthy volunteers. Number of CECs was measured by flow cytometry, and operation time was recorded. Changes in various indexes of patients were monitored, and diagnosis of ARDS was determined based on ARDS Berlin definition. We comprised d2 CECs in different groups, correlation between operation time and d2 CECs, ARDS of different severity by d2 CECs, and predictive value of d2 CECs for ARDS in postoperative patients.RESULTSThe number of d2 CECs in the ARDS group was significantly higher than that in the healthy control group (P < 0.001). The number of d2 CECs in the ARDS group was significantly higher than that in the non-ARDS group (P < 0.001). The number of d2 CECs in the non-ARDS group was significantly higher than that in the healthy control group (P < 0.001). Operation time was positively correlated with number of CECs on d2 (rs = 0.302, P = 0.001). The number of d2 CECs in the deceased group was significantly higher than that in the improved group (P < 0.001). There was no significant difference in number of d2 CECs between patients with mild and moderate ARDS. The number of d2 CECs in patients with severe ARDS was significantly higher than that in patients with mild and moderate ARDS (P = 0.041, P = 0.037). There was no significant difference in number of d5 and d2 CECs in the non-ARDS group after admission to intensive care. The number of d5 CECs was higher than the number of d2 CECs in the ARDS improved group (P < 0.001). The number of d5 CECs was higher than the number of d2 CECs in the ARDS deceased group (P = 0.002). If the number of CECs was > 1351/mL, sensitivity and specificity of predicting ARDS were 80.8% and 78.1%, respectively. CONCLUSIONChanges in number of CECs might predict occurrence and adverse outcome of ARDS after surgery, and higher numbers of CECs indicate worse prognosis of ARDS.

Патогенетическое значение дисфункции эндотелия в формировании гипертонуса периферической сосудистой стенки при местной холодовой травме

Актуальность. В структуре общего травматизма местная холодовая травма имеет большой удельный вес. Выявление новых маркеров криоповреждения способствует более ранней диагностике глубины поражений тканей при обморожении, созданию новых подходов к хирургическому лечению и ускоренной реабилитации пострадавших. Цель исследования - изучение в динамике содержания оксида азота, асимметричного диметиларгинина и циркулирующих эндотелиальных клеток в сыворотке крови у пациентов с местной холодовой травмой. Методика. В исследование включено 80 пациентов с обморожениями нижних конечностей III-IV степени в позднем реактивном периоде и периоде гранулирования и эпителизации. Использован мультиплексный анализ сыворотки крови набором реагентов фирмы Biomedical (США), методом Hladovec (1978) и методом П.П. Голикова (2004) соответвенно. Результаты. У пациентов с криотравмой во всех исследуемых группах установлено снижение уровня оксида азота относительно контроля. У пострадавших с местной холодовой травмой на 5-е сут отмечено значительное снижение в крови уровня асимметричного диметиларгинина. на 30-е сут с момента криоповреждения уровень асимметричного диметиларгинина не отличался от показателей контрольной группы. При обморожении дистальных сегментов стоп концентрация асимметричного диметиларгинина в крови не менялась относительно контроля. У пациентов с тяжелыми обморожениями нижних конечностей уровень асимметричного диметиларгинина в крови снижался в 3,6 раза. У пациентов с местной холодовой травмой на 5-е сут установлено увеличение в крови уровня циркулирующих эндотелиальных клеток в 5,2 раза, на 30-е сут с момента криоповреждения уровень циркулирующих эндотелиальных клеток снижался, но, по-прежнему, превышал контрольные значения. У пострадавших с обморожением дистальных сегментов стоп содержание в крови циркулирующих эндотелиальных клеток увеличилось в 2,7 раза, с поражением проксимальных сегментов нижних конечностей - в 6,7 раза относительно здоровых добровольцев. У пациентов с наиболее тяжелыми обморожениями нижних конечностей уровень циркулирующих эндотелиальных клеток в крови повышался в 9 раз. The relevance of local cold injury remains high. In the structure of general injury prevalence, proportion of freeze burns is great. Identification of new markers of cold injury will help earlier determination of tissue lesion depth, development of new approaches in the surgical treatment of deep freeze burns of extremities, and promote rehabilitation of patients. The aim of this study was to elucidate the dynamics of the serum content of nitric oxide (NO), asymmetric dimethylarginine (ADMA), and circulating endothelial cells (CEC) in patients with local cold injury. Methods. The study included 80 patients with III-IV degree frostbite of the lower extremities in the late reactive period and the period of granulation and epithelization. Multiplex analyses of blood serum were performed with Biomedical (USA) reagent kits, according to methods by Hladovec (1978) and Golikov (2004). Results. In all groups of patients with frostbite, NO level was decreased compared to the control. Patients with local cold trauma displayed significant decreases in blood levels of ADMA on day 5; on day 30 after the cold trauma, the ADMA level did not differ from the control group. In patients with frostbite of the distal foot segments, the blood concentration of ADMA did not change from the control level. In patients with the most severe frostbite of the lower extremities, serum ADMA decreased 3.6 times. Patients with local cold trauma had a 5.2-fold increase in the blood level of CEC on day 5. On day 30 after the freeze burn, the amount of CEC was decreased but still remained above the control value. In patients with frostbite of the distal foot segments, the content of CEC increased 2.7 times compared to the control. In patients with lesions of more proximal segments of the lower extremities, the CEC index was increased 6.7 times compared to healthy volunteers. In patients with the most severe frostbite of the lower extremities, the blood content of CEC was 9 times increased.

The Effect of Garcinia Mangostana Linn Extract to The Levels of Circulating Endothelial Cells and Endothelial Progenitor Cells in Patients with High Framingham Score

Background : Recently, studies have concerned on the use of xanthones for treating patients with cardiovascular diseases. In our country, xanthones were found in Garcinia Mangostana Linn. Objectives : To assess the effect of Garcinia Mangostana Linn Extract (GMLE) to the levels of Circulating Endothelial Cells (CEC) and Endothelial Progenitor Cells (EPC) in patients with high framingham score. Methods : A prospective cohort study was conducted from November 2018 to January 2019. The patients were divided into two groups. The first group was given 2520 mg/day of GMLE for 90 days and the second group was given placebo. In sub group analyses, patients were divided based on previous medication, including statin, statin and angiotensin-converting-enzyme inhibitors (ACEI) or angiotensin II receptor blockers (ARB), statin and oral antidiabetic drugs (OAD), and statin and ACEI or ARB and OAD. The outcome measures were CEC and EPC levels, measured at baseline and after 90 days of treatment. We used multiple linear regression to analyze the correlation and effect estimates. Results: A total of 72 patients was included in our study. All of the patients baseline characteristics were distributed homogeneously (p&gt;0.05). Our findings confirmed that GMLE administration was associated with decreased CEC level compared to placebo. On other hands, increased EPC level compared to placebo was observed after GMLE administration. In sub-group analyses, our study found that the combination of GMLE with statin and ACEI or ARB and the combination of GMLE with statin and ACEI or ARB and OAD were associated with decreased level of CEC compared to placebo, with the odd ratios were 0.12 and 0.18, respectively. Conversely, increased level of EPC was observed in subjects receiving the combination of GMLE with statin and ACEI or ARB and the combination of GMLE with statin. Conclusion: Administration of GMLE as adjuvant therapy is associated with the improvement of CEC and EPC levels in patients with high framingham scores.

Multimodality liquid biopsy for early monitoring and outcome prediction in first-line metastatic HER2-negative breast cancer: Final results of the prospective cohort from the French Breast Cancer InterGroup Unicancer (UCBG)— COMET study.

3019 Background: Circulating Tumor Cells (CTC) are independent markers of progression-free survival (PFS) and overall survival (OS) in patients (pts) with metastatic breast cancer (MBC). Monitoring circulating tumor DNA (ctDNA) can detect mutation associated with resistance to treatment and its variations reflect changes in tumor burden. We prospectively monitored CTC, Circulating Endothelial Cells (CEC), serum markers and ctDNA during first line chemotherapy for MBC. Methods: The French cohort COMET is a prospective study including first line HER2 negative pts receiving weekly paclitaxel and bevacizumab . Blood samples were obtained at baseline (BL) and before the second cycle of chemotherapy (C2).We present here the final planned analysis. Results: From 09/2012 to 11/2014, 286 patients were included: 198 for ctDNA, 251 for CEC and 283 for CTC. Median age was 56 years and 23% of pts had triple negative BC. At baseline, 71% of pts had ≥1 detectable CTC per 7.5 ml of blood (median 4 CTC, range 1- 30,000). With a threshold of ≥5 CTC, 49% of pts were positive at baseline and 22% at C2. For ctDNA, out of the first 196 pts analyzed, 147 had at least one somatic mutation (SNV) detected in plasma (75%). The average number of mutations per pt was 2.4 (range 1 to 9). Most commonly mutated genes were TP53 and GATA3. ESR1 was mutated in 10.6% of the pts and restricted to the ER+ subgroup. PIK3CA was mutated in 23.2% of the pts. Median Allelic Frequency was 9.1% . Only 68 pts (36%) had detectable ctDNA at C2. At baseline, CTC and ctDNA levels were correlated (r = 0.40, p &lt; 0.0001). Despite no complete overlap, 24 pts (12%) had no CTC nor ctDNA detected at baseline. Median follow-up was 53 months and median OS was 32 months. Detectable CTC and ctDNA at baseline and at C2 were significantly associated with decreased PFS and OS. CEC and serum markers level had no prognostic value. At multivariate analysis, triple negative status, detectable ctDNA at C2, CTC ≥5 at C2 and grade 3 on primary tumor were independent prognostic factors. Conclusions: This is the largest prospective cohort assessing the respective prognostic values of early CTC and ctDNA changes in homogenously treated first line MBC pts. Early decrease of CTC and ctDNA after one cycle of chemotherapy are independent predictive markers of favorable outcome, with a stronger value for ctDNA compared to CTC. Clinical utility of early ctDNA variations monitoring and changes in mutation profile remain to be demonstrated. Clinical trial information: NCT01745757.

Circulating endothelial cells in patients with alcohol withdrawal syndrome (uncomplicated and complicated delirium)

To study the content of circulating endothelial cells in the blood of patients with alcohol withdrawal syndrome (uncomplicated and complicated delirium). Thirty patients, aged from 18 to 40 years, with the diagnosis of 'alcohol withdrawal, uncomplicated' (F10.302) or 'alcohol withdrawal complicated by delirium' (F10.40) were examined in the acute state and after 2 weeks of therapy. The control group consisted of 22 normals. The content of circulating endothelial cells (CEC) in the blood was measured by flow-cytometry. Significant changes in the CEC content were observed in alcohol withdrawal syndrome: a multiple increase in the number of CECs due to the CEC-Living subpopulation, the appearance of CEC-Dead, which characterized the pronounced damage to the endothelium; the compensatory growth of ECP (CD146+CD34+) and AEC (CD146+CD105+). Patients with uncomplicated abstinence after 2 weeks of therapy showed positive changes in the form of a decrease in CEC level due to the CEC-Living. The CEC level remained high in patients with alcohol delirium after 2 weeks of treatment, the decrease in the content CEC-Living with a significant growth CEC-Dead and a slight compensatory increase in ECP (CD146+CD34+) was observed.

Circulating tumour cells and pathological complete response: independent prognostic factors in inflammatory breast cancer in a pooled analysis of two multicentre phase II trials (BEVERLY-1 and -2) of neoadjuvant chemotherapy combined with bevacizumab

We present a pooled analysis of predictive and prognostic values of circulating tumour cells (CTC) and circulating endothelial cells (CEC) in two prospective trials of patients with inflammatory breast cancer (IBC) treated with neoadjuvant chemotherapy combined with neoadjuvant and adjuvant bevacizumab. Nonmetastatic T4d patients were enrolled in two phase II multicentre trials, evaluating bevacizumab in combination with sequential neoadjuvant chemotherapy of four cycles of FEC followed by four cycles of docetaxel in HER2-negative tumour (BEVERLY-1) or docetaxel and trastuzumab in HER2-positive tumour (BEVERLY-2). CTC and CEC were detected in 7.5 and 4 ml of blood, respectively, with the CellSearch System. From October 2008 to September 2010, 152 patients were included and 137 were evaluable for CTC and CEC. At baseline, 55 patients had detectable CTC (39%). After four cycles of chemotherapy, a dramatic drop in CTC to a rate of 9% was observed (P < 0.01). Pathological complete response (pCR) rate was 40%. No correlation was found between CTC or CEC levels and pCR rate. Median follow-up was 43 months. CTC detection (≥1 CTC/7.5 ml) at baseline was associated with shorter 3-year disease-free survival (39% versus 70% for patients without CTC, P < 0.01, HR 2.80) and shorter 3-year overall survival (OS) (P < 0.01). In multivariate analysis, independent prognostic parameters for shorter survival were absence of hormonal receptors, no pCR and CTC detection at baseline. CEC level at baseline or variations during treatment had no prognostic value. In this pooled analysis of two prospective trials in nonmetastatic IBC, detection rate of CTC was 39% with a strong and independent prognostic value for survival. Combination of pCR after neoadjuvant treatment with no CTC detection at baseline isolated a subgroup of IBC with excellent OS (94% 3-year OS), suggesting that CTC count could be part of IBC stratification in prospective trials.

A pilot study using dynamic contrast enhanced-MRI as a response biomarker of the radioprotective effect of memantine in patients receiving whole brain radiotherapy.

PurposeThis pilot prospective study sought to determine whether dynamic contrast enhanced MRI (DCE-MRI) could be used as a clinical imaging biomarker of tissue toxicity from whole brain radiotherapy (WBRT).Method14 patients who received WBRT were imaged using dynamic contrast enhanced DCE-MRI prior to and at 8-weeks, 16-weeks and 24-weeks after the initiation of WBRT. Twelve of the patients were also enrolled in the RTOG 0614 trial, which randomized patients to the use of placebo or memantine. After the unblinding of the treatments received by RTOG 0614 patients, DCE-MRI measures of tumor tissue and normal appearing white matter (NAWM) vascular permeability (Initial Area Under the Curve (AUC) Blood Adjusted) was analyzed. Cognitive, quality-of-life (QOL) assessment and blood samples were collected according to the patient's ability to tolerate the exams. Circulating endothelial cells (CEC) were measured using flow cytometry.ResultsFollowing WBRT, there was an increasing trend in the vascular permeability of tumors (p=0.09) and NAWM (p=0.06) with time. Memantine significantly (p=0.01) reduced NAWM AUC changes following radiotherapy. Patients on memantine retained (COWA p= 0.03) better cognitive functions than those on placebo. No association was observed between the level of CEC and DCE-MRI changes, time from radiotherapy or memantine use.ConclusionsDCE-MRI can detect vascular damage secondary to WBRT. Our data suggests that memantine reduces WBRT-induced brain vasculature damages.

A Phase II Clinical Trial of TRC105 (Anti-Endoglin Antibody) in Adults With Advanced/Metastatic Urothelial Carcinoma

BackgroundIn this trial we assessed the efficacy and tolerability of TRC105, a chimeric monoclonal antibody that targets CD105 (endoglin) in patients with advanced, previously treated urothelial carcinoma (UC). Patients and MethodsPatients received TRC105 15 mg/kg every 2 weeks on days 1 and 15 of each 28-day cycle. The primary end point was progression-free survival (PFS) at 6 months. Secondary end points included safety, toxicity, and overall survival (OS). CD105 expression was evaluated using immunohistochemistry (IHC) in a separate cohort of 50 UC patients. Biomarker studies included immune subsets, circulating tumor cells (CTCs), circulating endothelial cells (CECs), circulating endothelial progenitor cells (CEPs), and osteopontin. ResultsOf 13 patients enrolled, 12 were evaluable for OS and PFS. The 3-month PFS probability was 18.2% (median PFS, 1.9 months [95% confidence interval (CI), 1.8-2.1 months). This met the criterion for ending accrual on the basis of the 2-stage design. Median OS was 8.3 months (95% CI, 3.3-17.0 months). IHC for CD105 scores was not associated with T stage (P = .26) or presence of lymph nodes (P = .64). Baseline levels of regulatory T and B cells, CEPs, and changes in CEC level after TRC105 exhibited trends toward an association with PFS or OS. CTCs pre- and post-TRC105 were detected in 4 of 4 patients. ConclusionAlthough TRC105 was well tolerated, it did not improve 6-month PFS in heavily pretreated patients with advanced UC. CD105 staining was present in 50% of UC tumors at different intensities. Our observations on the pharmacodynamic significance of immune subsets, CECs, and CTCs warrant further study.

Circulating Endothelial Cells and Endothelial Function Predict Major Adverse Cardiac Events and Early Adverse Left Ventricular Remodeling in Patients With ST-Segment Elevation Myocardial Infarction.

Endothelial progenitor cells (EPCs) and circulating endothelial cells (CECs) are mobilized from the bone marrow and increase in the early phase after ST-elevation myocardial infarction (STEMI). The aim of this study was to assess the prognostic significance of CECs and indices of endothelial dysfunction in patients with STEMI. In 78 patients with acute STEMI, characterization of CD34+/VEGFR2+CECs, and indices of endothelial damage/dysfunction such as brachial artery flow mediated dilatation (FMD) were determined. Blood samples for CECs assessment and quantification were obtained within 24 hours of admission and FMD was assessed during the index hospitalization. At 30 days follow up, the primary composite end point of major adverse cardiac events (MACE) consisting of all-cause mortality, recurrent nonfatal MI, or heart failure and the secondary endpoint of early adverse left ventricular (LV) remodeling were analyzed. The 17 patients (22%) who developed MACE had significantly higher CEC level (P = 0.004), von Willebrand factor (vWF) level (P = 0.028), and significantly lower FMD (P = 0.006) compared to the remaining patients. Logistic regression analysis showed that CECs level and LV ejection fraction were independent predictors of MACE. The areas under the receiver operating characteristic curves (ROC) for CEC level, FMD, and the logistic model with both markers were 0.73, 0.75, and 0.82, respectively, for prediction of the MACE. The 16 patients who developed the secondary endpoint had significantly higher CEC level compared to remaining patients (P = 0.038). In conclusion, increased circulating endothelial cells and endothelial dysfunction predicted the occurrence of major adverse cardiac events and adverse cardiac remodeling in patients with STEMI.

Prognostic value of circulating endothelial cells in non-small cell lung cancer patients: a systematic review and meta-analysis.

Circulating endothelial cells (CECs) have been indicated as a potential biomarker of vascular damage in a variety of cancers. Several studies have revealed CECs may reflect the extent of tumor angiogenesis; however, the role of CECs in the prognosis of non-small cell lung cancer (NSCLC) is undetermined to date. A meta-analysis has been prepared to determine whether the base level of CECs and the changes of CECs after therapy (∆CECs: post-therapeutic value minus the pre- therapeutic value) could be considered as a prognostic tool for patients with NSCLC. Systematic reviews of studies published before April 30 2015 were conducted on the association between the levels of CECs or ∆CECs and the prognosis of NSCLC in several data bases. Hazard ratios (HRs) and the 95% confidence intervals (CIs) were used to collate the data. Similarly, heterogeneity and publication bias were also evaluated. A total of nine studies, containing eight prospective studies and one retrospective study, involving 515 patients was identified. Patients with higher level of CECs counts at baseline were associated with longer progression-free survival (PFS) (HR 0.71, 95% CI: 0.529-0.891). ∆CECs could also be considered a prognostic indicator in NSCLC patients (HR 0.575, 95% CI: 0.401-0.75). The former and the later are without a significant heterogeneity in the data (I(2)=21.2% and 0.0%, P=0.274 and 0.870, respectively). However, there was no correlation between the base level of CECs and the overall survival (OS) (HR 0.914, 95% CI: 0.560-1.267, I(2)=43.6%, P=0.150). Higher levels of CECs counts at baseline and the ensuing decrease after therapy demonstrated a positive correlation with longer PFS in NSCLC patients. But this phenomenon has not been found in OS. From a certain perspective, CECs counts and ∆CECs could be potential prognostic indicators for NSCLC patients.

Meta-Analysis of Circulating Endothelial Cells and Circulating Endothelial Progenitor Cells as Prognostic Factors in Lung Cancer.

The aim of this study was to analyze the prognostic implications of pretreatment circulating endothelial cells (CECs) and circulating endothelial progenitor cells (CEPCs) for the survival of patients with lung cancer. Relevant literature was identified using Medline and EMBASE. Patient clinical characteristics, overall survival (OS) and progression-free survival (PFS) together with CEC and CEPC positive rates before treatment were extracted. STATA 12.0 was used for our analysis and assessment of publication bias. A total of 13 articles (8 for CEC and 5 for CEPC, n=595 and n=244) were pooled for the global meta-analysis. The odds ratio (OR) for OS predicted by pretreatment CECs was 1.641 [0.967, 2.786], while the OR for PFS was 1.168 [0.649, 2.100]. The OR for OS predicted by pretreatment CEPCs was 12.673 [5.274, 30.450], while the OR for PFS was 4.930 [0.931, 26.096]. Subgroup analyses were conducted according to clinical staging. Odds ratio (OR) showed the high level of pretreatment CECs only correlated with the OS of patients with advanced lung cancer (stage III-IV). High counts of CECs seem to be associated only with worse 1-year OS in patients with lung cancer, while high level of pretreatment CEPCs correlate with both worse PFS and OS.

Abstract 381: Circulating tumor cells (CTC) but not circulating endothelial cells (CEC) are independent prognostic factors in neoadjuvant chemotherapy combined with bevacizumab in HER2 negative inflammatory breast cancer (IBC) in multicentre phase II trial BEVERLY1

Abstract Background: CTC detection in blood is an independent prognostic factor in primary breast cancer with a detection rate from 20 to 25% (Pierga, CCR 2008, Rack B, JNCI 2014). Higher incidence of 35% of CTC has been reported in IBC (Pierga, Lancet Oncol 2012). Predictive and prognostic value of CEC for response to anti-angiogenic agents remains debated. Methods: CTC and CEC were detected in 7.5 ml and 4 ml of blood respectively in the neoadjuvant setting of HER2 negative IBC (T4d) patients (pts) enrolled in the phase II multicentre trial BEVERLY 1, evaluating bevacizumab (15mg/kg q3w) in combination with sequential neoadjuvant CT of 4 cycles of FEC followed by 4 cycles of Docetaxel. The CellSearch™ System, combining EpCAM immunomagnetic selection followed by anti-cytokeratin (A45B/B3) and anti-HER2 staining for CTC and CD146 IMS and CD105 staining for CEC. Results: From 12/08 to 09/10, 101 pts were included and 92 were evaluable for CTC and CEC. At baseline, 37 pts out of 92 had ≥ 1 detectable CTC (40%, 95%CI 30-50%). No correlation was found between CTC and CEC levels. A dramatic drop in CTC incidence (p&amp;lt;0.0001) was observed from baseline to the 1st follow-up analysis after 4 cycles (6%). There was a significant increase of CEC median number from baseline (12) to preoperative sample (130) (p &amp;lt;0.001) and a decrease (39) (p = 0.04) when CT and bevacizumab were interrupted for surgery. CTC detection at baseline independently predicted 3-year DFS: (73% vs. 41% for patients with &amp;lt;1 vs. ≥1 CTC/7.5 mL [p&amp;lt;0.001, HR 2.73 (1.48-5.04)]) and OS (94% vs.51% [p&amp;lt;0.001, HR 5.93 (2.49-14.10)]. Fourteen patients (15%) with no CTC detected at baseline and with complete pathological response (pCR) had a high 3-year DFS (93%) and OS (100%). CEC level under 20/4ml at baseline did not reach statistical significance as a favorable prognostic factor for DFS and OS (p = 0.055 and 0.066 respectively). CEC changes during treatment had no prognostic value. Conclusions: We observed a high CTC detection rate of 40% in pts with HER2 negative IBC, including triple negative tumor, with a strong and independent prognostic value for DFS and OS. Combining pCR after neoadjuvant chemotherapy combined with bevacizumab, and CTC at baseline, isolates a subgroup of inflammatory breast cancer with excellent survival. Citation Format: Jean-Yves Pierga, Francois-Clement Bidard, Aurelie Autret, Fabrice Andre, Thierry Petit, Florence Dalenc, Christel Levy, William Jacot, Jacques Bonneterre, Jean-Marc Ferrero, Pierre Kerbrat, Jerome Lemonnier, Francois Bertucci, Patrice Viens. Circulating tumor cells (CTC) but not circulating endothelial cells (CEC) are independent prognostic factors in neoadjuvant chemotherapy combined with bevacizumab in HER2 negative inflammatory breast cancer (IBC) in multicentre phase II trial BEVERLY1. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 381. doi:10.1158/1538-7445.AM2015-381

Predictive and prognostic significance of circulating endothelial cells in advanced non-small cell lung cancer patients.

The aim of this study was to evaluate the predictive and prognostic values of circulating endothelial cells (CECs) in patients with advanced non-small cell lung cancer (NSCLC). A total of 102 newly diagnosed advanced NSCLC patients were enrolled in this study. The amount of CECs was enumerated by flow cytometry (CD45- CD31+ CD146+) at baseline. CEC counts of 56 patients were detected before and after two cycles of chemotherapy. We correlated the baseline and reduction of CECs after therapy with objective response rate (ORR), progression-free survival (PFS), and overall survival (OS). The CEC level was significantly higher in advanced NSCLC patients, ranging from 57 to 1300 cells/10(5) cells (mean ± SD = 299 ± 221 cells/10(5) cells), than in patients with benign lesions (205 ± 97 cells/10(5) cells) and healthy volunteers (117 ± 33 cells/10(5) cells). When the cutoff value of CEC counts was 210 cells/10(5) cells, there was no significant association between CEC counts and OR/PFS/OS of the enrolled patients. However, patients with CEC response after chemotherapy have more chances to achieve OR (P < 0.001), and such patients showed longer PFS (P = 0.048) and OS (P = 0.018) than those without CEC response. In the multivariate analysis, the independent prognostic roles of brain metastasis (HR 6.165, P = 0.001), and CEC response (HR 0.442, P = 0.044) were found. The CEC counts could be considered as diagnostic biomarker for advanced NSCLC patients. And the reduction of CECs after treatment might be more ideal than the baseline CEC counts as a predictive or prognostic factor in patients treated with chemotherapy or anti-angiogenic therapy.

Circulating endothelial cells and vascular cell adhesion molecule-1 correlates with severity of regurgitation lesion and heart failure in rheumatic heart disease

Objective: This study aims to determine the correlation of vascular cell adhesion molecule-1 (VCAM-1) and circulating endothelial cells (CEC) with cardiac valves lesion severity and heart failure (HF) in rheumatic heart disease (RHD). Methods: A cross-sectional observational study using human peripheral blood samples of 36 children aged 6 to14 years old, divided into two groups: 18 RHD patients and 18 healthy controls group. The expression of CECs and VCAM-1 on CECs was investigated by using flow-cytometry method while the levels of soluble VCAM-1 (sVCAM-1) were obtained by using ELISA method. The severity of valve lesions was determined in a qualitative manner. Results: The average of CECs expression, VCAM-1 on CECs expression, and sVCAM-1 levels were significantly higher in the patient group than those of the healthy group. The expression of VCAM-1 on CECs was significantly different among mild, moderate, and severe regurgitation. Correlation analysis showed that the CECs expression had a significant correlation with HF. Moreover, correlation was found between expression of VCAM-1 on CECs and the severity of valve lesions. However, sVCAM-1 in plasma had no correlation either with HF or with valve lesion. Conclusion: The expression of VCAM-1 on CECs is associated with severe regurgitation while CECs level was correlated with HF. VCAM-1 on CECs may be considered a marker for the severity of functional regurgitation in RHD. CECs could be recommended as a measure for RHD with HF.

Circulating endothelial cells and chronic kidney disease.

Endothelial dysfunction may play a crucial role in initiation of the pathogenesis of vascular disease and atherosclerosis. The identification and quantification of circulating endothelial cells (CEC) have been developed as a novel marker of endothelial function. We describe, in great detail, mechanisms of endothelial dysfunction and CEC detachment. We also review the relationship between numbers of CEC and disease severity and response to treatment. In addition, we describe the possible clinical use of CEC in chronic kidney disease (CKD) and kidney transplantation. In summary, CEC have been developed as a novel approach to assess the endothelial damage. Measurement of the CEC level would provide an important diagnostic and prognostic value on the endothelium status and the long-term outcome of vascular dysfunction.

Circulating endothelial cell levels in psoriatic patients and their modification after an anti‐TNF‐alpha (Etanercept) treatment

Endothelial function in psoriatic patients has been mainly evaluated through a high-resolution ultrasound measurement of flow-mediated vasodilation in the brachial artery, which is an operator-dependent and technically demanding technique: this characteristic, together with different patient selection criteria, could account for the conflicting results emerging from different studies. Recently, Circulating Endothelial Cells (CECs) level has been suggested as a novel biomarker of vascular injury. The number of CECs was determined by a semi-automated immunomagnetic system (CellSearch system) in peripheral blood of psoriatic patients (n = 48) and healthy subjects (n = 50). In 15 patients, CEC level was also evaluated after 6 months of treatment with an anti-TNF-alpha agent, Etanercept. The plasma levels of high-sensitivity C-reactive Protein (CRP), E-selectin, VEGF and PAI-1 were measured by ELISA. The psoriasis severity was assessed by PASI score. A statistically significant difference (P = 0.001) was found between CEC level in psoriatic patients (10.6 ± 9.4 cells/mL) vs. the control group (3.9 ± 0.9 cells/mL). This count inversely correlated with sE-selectin levels (r(2) = 0.16; P = 0.03). After 6 months of therapy, patients experienced a significant (P < 0.05) decrease in CEC levels (3.4 ± 1.3 cells/mL) and in PASI score (from 11.7 ± 8.1 to 2.1 ± 4.0). The elevated CECs level that we found in a sample of high selected psoriatic patients could be expression of endothelial damage. Lowering of CECs count after treatment with Etanercept support the hypothesis that an effective systemic therapy of psoriasis may also improve the endothelial function.