Molecularly determined total tumour load in lymph nodes of stage I–II colon cancer patients correlates with high-risk factors. A multicentre prospective study

Stage I–II (pN0) colorectal cancer patients are surgically treated although up to 25 % will eventually die from disease recurrence. Lymph node (LN) status is an independent prognostic factor in colorectal cancer (CRC), and molecular tumour detection in LN of early-stage CRC patients is associated with an increased risk of disease recurrence and poor survival. This prospective multicentre study aimed to determine the relationship between LN molecular tumour burden and conventional high-risk factors in stage I–II colon cancer patients. A total of 1940 LN from 149 pathologically assessed pN0 colon cancer patients were analysed for the amount of tumour cytokeratin 19 (CK19) messenger RNA (mRNA) with the quantitative reverse transcription loop-mediated isothermal amplification molecular assay One-Step Nucleic Acid Amplification. Patient’s total tumour load (TTL) resulted from the sum of all CK19 mRNA tumour copies/μL of each positive LN from the colectomy specimen. A median of 15 LN were procured per case (IQR 12;20). Molecular positivity correlated with high-grade (p < 0.01), mucinous/signet ring type (p = 0.017), male gender (p = 0.02), number of collected LN (p = 0.012) and total LN weight per case (p < 0.01). The TTL was related to pT stage (p = 0.01) and tumour size (p < 0.01) in low-grade tumours. Multivariate logistic regression showed independent correlation of molecular positivity with gender, tumour grade and number of fresh LN [AUC = 0.71 (95 % CI = 0.62–0.79)]. Our results show that lymph node CK19 mRNA detection correlates with classical high-risk factors in stage I–II colon cancer patients. Total tumour load is a quantitative and objective measure that may help to better stage early colon cancer patients.Work supported by the Banc de Tumors-Biobanc Hospital Clinic-IDIBAPS and Xarxa de Bancs de Tumors de Catalunya (XBTC), and by grants from the Fundación Científica de la Asociación Española Contra el Cáncer (GCB13131592CAST), Ministerio de Economía y Competitividad (SAF2014–54,453-R), Agència de Gestió d’Ajuts Universitaris i de Recerca (2014SGR135), and by Sysmex Coorp Spain (Sant Just Desvern, Spain). CIBERehd is funded by the Instituto de Salud Carlos II

Axillary staging based on molecular analysis: Results of the B-CLOSER-II study

Introduction Axillary staging (pN) is a strong predictor of outcome in early stage breast cancer yet following the publication of the Z0011 trial there has been an increasing tendency to spare lymph node dissection. Automated molecular detection of cytokeratin 19mRNA by one-step nucleic acid amplification (OSNA) has been demonstrated to be an accurate method to assess sentinel lymph node (SLN) metastasis. In this study we compare histological and molecular methods following complete axillary lymph node dissection (cALND), determine whether molecular axillary staging affects survival, and evaluate the predictive and prognostic value of total tumor load in ALND (AD-TTL) and in all positive nodes (G-TTL). Material and methods Axillary lymph nodes were collected from 102 patients with primary breast cancer with histological confirmation of axillary involvement (cN+) or positive SLN. The central 1-mm portion of each non-SLN was processed for hematoxylin-eosin staining and the remaining tissue was analyzed by OSNA. Results Non-SLNs were diagnosed as positive in 72 out of 102 patients (70.6 %) on OSNA compared with only 53 (52 %) on histology (p < 0.01). Thirteen patients would have changed staging if the diagnoses provided had been by molecular methods (p < 0.01), but without a change in prognosis. AD-TTL and G-TTL were predictive of recurrence and mortality. Conclusions Compared to molecular detection, histological examination significantly underestimates the frequency of axillary node metastases. However, the increase in pN did not show a clinical effect on survival in those patients.This study was supported by a Grant from Sysmex España S.L. The sponsor had no role in the study design, analysis, or interpretation of the data

Role of total tumour load of sentinel lymph node on survival in early breast cancer patients.

Axillary staging (pN) is considered one of the most important prognostic factors in breast cancer patients. However, the Z0011 study data drastically reduced the number of surgical axillary dissections in a selected group of patients, limiting the prognostic information relating to axillary involvement to the sentinel lymph node (SLN). It is known that there is a relationship between SLN total tumour load (TTL) and axillary involvement. The objective of this study is to analyse the relationship between the TTL and outcomes in patients with early stage breast cancer. clinicopathological and follow-up data were collected from 950 patients with breast cancer between 2009 and 2010 on whom SLN analysis was conducted by molecular methods (One Step Nucleic Acid Amplification, Sysmex, Kobe, Japan). TTL (defined as the total number of CK19 mRNA copies in all positive SLN) correlates with disease free survival (HR, 1.08; p = 0.000004), with local recurrence disease free survival (HR = 1.07; p = 0.0014) and overall survival (HR: 1.08, p = 0.0032), clearly defining a low-risk group (TTL 2.5 × 104 CK 19 mRNA copies/μL). SLN TTL permits the differentiation between two patient groups in terms of DFS and OS, independently of axillary staging (pN), age and tumour characteristics (size, grade, lymphovascular invasion). This new data confirms the clinical value of low axillary involvement and could partially replace the information that staging of the entire axilla provides in patients on whom no axillary lymph node dissection is performed

Elaboration of a nomogram to predict nonsentinel node status in breast cancer patients with positive sentinel node, intraoperatively assessed with one step nucleic amplification: Retrospective and validation phase

Background: Tumor-positive sentinel lymph node (SLN) biopsy results in a risk of non sentinel node metastases in micro-and macro-metastases ranging from 20 to 50%, respectively. Therefore, most patients underwent unnecessary axillary lymph node dissections. We have previously developed a mathematical model for predicting patient-specific risk of non sentinel node (NSN) metastases based on 2460 patients. The study reports the results of the validation phase where a total of 1945 patients were enrolled, aimed at identifying a tool that gives the possibility to the surgeon to choose intraoperatively whether to perform or not axillary lymph node dissection (ALND).Methods: The following parameters were recorded: Clinical: hospital, age, medical record number; Bio pathological: Tumor (T) size stratified in quartiles, grading (G), histologic type, lymphatic/vascular invasion (LVI), ER-PR status, Ki 67, molecular classification (Luminal A, Luminal B, HER-2 Like, Triple negative); Sentinel and non-sentinel node related: Number of NSNs removed, number of positive NSNs, cytokeratin 19 (CK19) mRNA copy number of positive sentinel nodes stratified in quartiles. A total of 1945 patients were included in the database. All patient data were provided by the authors of this paper.Results: The discrimination of the model quantified with the area under the receiver operating characteristics (ROC) curve (AUC), was 0.65 and 0.71 in the validation and retrospective phase, respectively. The calibration determines the distance between predicted outcome and actual outcome. The mean difference between predicted/observed was 2.3 and 6.3% in the retrospective and in the validation phase, respectively. The two values are quite similar and as a result we can conclude that the nomogram effectiveness was validated. Moreover, the ROC curve identified in the risk category of 31% of positive NSNs, the best compromise between false negative and positive rates i.e. when ALND is unnecessary ( 31%).Conclusions: The results of the study confirm that OSNA nomogram may help surgeons make an intraoperative decision on whether to perform ALND or not in case of positive sentinel nodes, and the patient to accept this decision based on a reliable estimation on the true percentage of NSN involvement. The use of this nomogram achieves two main gools: 1) the choice of the right treatment during the operation, 2) to avoid for the patient a second surgery procedure

Una revisión sistemática de las recomendaciones diagnósticas y terapéuticas del panel de expertos en cáncer de origen desconocido

Cancer of Unknown Primary (CUP) is a metastatic cancer with confirmed histology of which the primary origin is unknown after to work up initial evaluation through a pathological clinical study, the analytical and imaging study. The diagnostic process includes the early obtaining of quality biopsy material. It includes its histological and immunohistochemical analysis: a study to determine the tumor line, an analysis of cytokeratins and a large battery of antibodies to confirm the specific origin of each possible tumor type. The development of molecular platforms has allowed improving the diagnosis of CUP, increasing the number of patients who can benefit from treatment with specific therapy, significantly increasing the survival and reducing the toxicity. However, the updated guidelines (NICE, ESMO, NCCN) emphasize that the impact on the clinical benefit of the specific treatment according to the results of the molecular platforms is still controversial.El Cáncer de Origen Desconocido (COD) es un cáncer metastásico con histología confirmada del cual se desconoce el origen primario después de realizar un estudio diagnóstico inicial mediante el estudio clínico patológico, el estudio analítico y de imagen. El estudio diagnóstico incluye la obtención precoz de material de biopsia de calidad. Incluye su análisis histológico e inmunohistoquímico: un estudio para determinar la estirpe tumoral, análisis de citoqueratinas y una batería amplia de anticuerpos para confirmar el origen específico. El desarrollo de plataformas moleculares ha mejorado su diagnóstico, incrementando el número de pacientes que se benefician del tratamiento con terapia específica, aumentando su supervivencia y reduciendo la toxicidad. Sin embargo, las guías actualizadas (NICE, ESMO, NCCN) destacan que el impacto en el beneficio clínico del tratamiento específico según los resultados de las plataformas moleculares es todavía controvertido

Mechanistic aspects of water oxidation catalyzed by organometallic iridium complexes

The reactions of three iridium water‐oxidation catalysts {[Cp*IrL1L2L3]Xn; 1: L1, L2 = 2,2‐bipyridine (bpy), L3 = Cl, n = 1, X = Cl; 2: L1, L2 = 2‐benzoylpyridine (bzpy), L3 = NO3; 3: L1 = L2 = L3 = H2O, n = 2, X = NO3; Cp* = pentamethylcyclopentadienyl} with cerium ammonium nitrate (CAN) and NaIO4 (sacrificial oxidants, SOs) have been studied by Clark electrode measurements (both in solution and in the gas phase), on‐line mass spectrometry, manometry and UV/Vis spectroscopy. Furthermore, cyclic voltammetry has been applied to evaluate the relative tendency of 1 and 2 to be oxidized. The turnover frequency (TOF) increases as the ratio (R) between the concentration of the SO and that of the catalyst increases. O2 production with CAN is observed in experiments with R = 20 for 1 and 3, whereas O2 becomes detectable with 2 only when R = 40. Catalyst 2 has the highest tendency to be oxidized to IrIV and forms a blue intermediate I characterized by a UV/Vis band at 574 nm. The formation of I occurs with the same velocity as that of the production of O2, which indicates that I is a species directly involved in the catalytic cycle. The disappearance of I, when O2 evolution is finished, is a second‐order process more than one order of magnitude slower than O2 production and is strongly accelerated by the presence of benzyl alcohol. This suggests that I is a molecular species that slowly undergoes disproportion when catalysis is over. Experiments in which multiple aliquots of SO (CAN) were added (R = 20 and 40) indicate that catalysts 1–3 can reinitiate the catalytic cycle once they have been kept in a dormant state for 0–9 min; the TOFs of the second and third additions are approximately equal and higher than that of the first addition. By combining manometry and on‐line mass spectrometry measurements, it was found that O2 evolution is parallel to the production of a small amount of CO2 owing to catalyst degradation. The TOFs of the experiments performed with NaIO4 as the SO are about 2–3 times lower than those with CAN, but the same reactivity order is found 3 > 2 > 1. The activation parameters were evaluated with NaIO4 for all catalysts and with CAN for 2 at 10–45 °C. ΔG# is practically the same in all situations (25–26 kcal mol–1), whereas ΔH# is appreciably lower for 2 (13.1 kcal mol–1 with CAN and 13.3 kcal mol–1 with NaIO4) than for 1 (16 kcal mol–1) and 3 (16.9 kcal mol–1). The lowest enthalpic cost with 2 is balanced by the highest entropic cost (–41 cal mol–1 K–1) that approaches that typical for an associative bimolecular process