Estudio de factores patológicos y moleculares con valor pronóstico y predictivo de respuesta en el carcinoma colo-rectal

Las metástasis ganglionares son un factor pronóstico clave en cáncer colorrectal, asociándose a una mayor recurrencia loco-regional así como una menor supervivencia a los 5 años. Entre los factores predictivos de respuesta se encuentran el estado mutacional de K-Ras (que selecciona los pacientes susceptibles de recibir tratamiento biológico adyuvante) y, en tumores de recto operados, la calidad resección del mesorrecto, la afectación del margen circunferencial y el grado de regresión en los casos tratados, variables que se relacionarán con recidivas y supervivencia. En 100 pacientes con cáncer de colon en estadio I-II hemos comprobado que el método OSNA (One-step Nucleic Acid Amplification) consigue un índice de supraestadificación del 25% de los casos, siendo un método fiable y reproducible, habiéndose diseñado un protocolo de identificación de ganglios centinela “ex vivo” con una alta especificidad. En 200 pacientes con cáncer colo-rectal metastático hemos constatado que las mutaciones 12Ser y 12Cys de K-Ras se asocian a una mayor mortalidad y que los tumores de alto grado o con una variedad histológica diferente al adenocarcinoma convencional ofrecen una peor supervivencia. En 200 piezas quirúrgicas de cáncer de recto, hay una asociación directa entre la calidad de resección de mesorrecto y la progresión de la enfermedad, así como la mortalidad de estos pacientes.Les metàstasis ganglionars són un factor pronòstic clau en càncer colo-rrectal, associant-se a una major recurrència loco-regional i a una pitjor supervivència als 5 anys. Entre els factors predictius de resposta es troben l’estat mutacional de K-Ras (que selecciona els pacients susceptibles de rebre tractament biològic adjuvant) i, en tumors de recte operats, la qualitat de resecció del mesorrecte, l’afectació del marge circumferencial i el grau de regresió en els casos tractats, variables que es relacionaran amb recidives i supervivència. En 100 pacients amb càncer de colon en estadi I-II hem comprovat que el mètode OSNA (Onestep Nucleic Acid Amplification) aconsegueix un índex de supraestadificació del 25% dels casos, essent un mètode fiable i reproduïble, havent-ne dissenyat un protocol d’identificació de ganglis sentinella “ex vivo” amb una alta especificitat. En 200 pacients amb càncer colo-rectal metastàsic hem constatat que les mutacions 12Ser i 12Cys de K-Ras s’associen a una major mortalitat i que els tumors d’alt grau o amb una varietat histològica diferent al adenocarcinoma convencional presenten una pitjor supervivència. En 200 peces quirúrgiques de càncer de recte, hi ha una associació directa entre la qualitat de resecció del mesorrecte i la progressió de la malaltia, així com la mortalitat d’aquests pacients.Lymph node metastasis is a key prognosis factor in colorectal carcinoma. It associates to local recurrence and to a low rate of survival in the first 5 years of follow-up. Among the predictive factors of tumor response are the K-Ras mutation, which if is it negative allows this patient to be treated with targeted antibodies, and, in rectal cancer, the quality of the mesorectal excision surgery, the involved circumferential margin and the tumor regression if is treated. These factors are related to recurrence and survival. In 100 colon cancer patients in stage I or II, the OSNA method (One-step Nucleic Acid Amplification) upstaged 25% of cases to stage III. OSNA is a feasible method and we have got an “ex vivo” protocol for identifying sentinel lymph nodes with a high rate of specificity. In 200 patients with metastatic colon cancer 12Ser and 12Cys mutations associate to higher level of mortality and either high grade tumors or variants of tumors other than conventional adenocarcinoma associate to poor survival rate. In 200 rectal cancer specimens there is an strong association between the quality of the mesorectal excision and cancer progression, and also with the survival rate

Cytology Smears: An enhanced alternative method for colorectal cancer pN Stage-A multicentre study

Stage II colorectal cancer (CRC) recurrence remains a clinical problem. Some of these patients are true stage III CRC with a pN0 pathology stage. This large prospective multicentre cohort study aimed at evaluating the diagnostic ability of lymph node (LN) cytology smears to perform the pN stage and compare it with the conventional haematoxylin and eosin (H&E) pathology pN stage. Additionally, we used the One-Step Nucleic Acid Amplification (OSNA), a high-sensitive molecular method of LN staging. A total of 3936 fresh LNs from 217 CRC surgical specimens were examined by three methods, H&E, LN cytology smears, and OSNA. H&E detected 29% of patients with positive LNs, cytology smears 35%, and OSNA 33.2% (p < 0.0001). H&E and cytology concordantly classified 92.2% of tumours, and 88.5% between OSNA and HΕ Cytology had 96.8% sensitivity and 90.3% specificity to discriminate positive/negative patients compared to H&E (p = 0.004), and 87.3% sensitivity and 89% specificity when compared to OSNA (p = 0.56). Patients with positive LNs detected by any of the three methods had significantly worse disease-free and overall survival. We conclude that pN stage accuracy for detecting positive LNs is superior with LN cytological smears than with conventional H&E, which would enable a better pN stage and management of early-stage CRC patients.This research was funded by Fondo de Investigación Sanitaria grant number PI17/01304, PI20/00863, awarded to MC and JC. We acknowledge the Agència de Gestió d’Ajuts Universitaris i de Recerca (Generalitat de Catalunya, GRC 2017SGR653,). This article is based upon work from COST Action CA17118, supported by COST (European Cooperation in Science and Technology). www.cost.eu. SL holds a PFIS grand from Instituto de Salud Carlos iii and co-funded by the European Regional Development Fund (ERDF) (FI18/00221)

Lymph Node Tumor Burden Correlates With Tumor Budding and Poorly Differentiated Clusters: A New Prognostic Factor in Colorectal Carcinoma?

Introduction: Molecular lymph node (LN) staging in early colorectal cancer (CRC) has demonstrated to be more precise than conventional histopathology pN staging. Tumor budding (TB) and poorly differentiated clusters (PDCs) are associated with LN metastases, recurrences, and lower survival in CRC. We evaluated the correlation between the total tumor load (TTL) in LNs from CRC surgical specimens with patient outcome, TB, and PDC. Methods: In this retrospective multicentre study, 5,931 LNs from 342 stage I-III CRC were analyzed by both hematoxylin and eosin and molecular detection of tumor cytokeratin 19 mRNA by one-step nucleic acid amplification. TB and PDC were evaluated by hematoxylin and eosin and cytokeratin 19 immunohistochemistry. Results: One-step nucleic acid was positive in 38.3% patients (n = 131). Tumor Budding was low in 45% cases, intermediate in 25%, and high in 30%. Poorly Differentiated Clusters were low-grade G1 in 53%, G2 in 32%, and G3 in 15%. TB and PDC correlated with TTL, high-grade, lymphovascular and perineural invasion, pT, pN and stage (P < 0.001). TB, PDC, and TTL ≥ 6,000 copies/µL were associated with worse overall survival (P = 0.002, P = 0.013, and P = 0.046) and disease-free survival (P < 0.001). Discussion: The implementation of more sensitive molecular methods to assess LN status is a promising alternative approach to pN staging, which could be integrated to other factors to help risk stratification and management of patients with early-stage CRC. This study demonstrates the correlation of the amount of LN tumor burden with TB and PDCs. TTL is related to the outcome and could be used as a new prognostic factor in CRC (see Visual Abstract, Supplementary Digital Content 2, http://links.lww.com/CTG/A512).This work was supported by the Instituto de Salud Carlos III (grants PI17/01304 to J.C and M.C; grant PI16/00766 to F.B., and a Miguel Servet grant to J.C, cofunded by the European Regional Development Fund (ERDF) (CPII18/00026), through the Plan Estatal de Investigacion Científica y Tecnica y de Innovación, and Agencia de Gestiò d’Ajuts Universitaris i de Recerca (2017SGR653 and 2017SGR1035). CIBERehd is funded by the Instituto de Salud Carlos III. We also acknowledge the support of the CERCA Programme/Generalitat de Catalunya, and the Xarxa de Bancs de Tumors de Catalunya (XBTC)

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

Factors associated with higher rate of Complete Pathologic Response after Long-Course Neoadjuvant Treatment for Locally Advanced Rectal Cancer Patients – Results from a retrospective cohort study focused on Inflammatory Indexes

BACKGROUND: Pathologic complete response (pCR) after neoadjuvant chemoradiotherapy (NCRT) has a prognostic value in locally advanced rectal cancer (LARC). This study aimed to evaluate the ability to predict pCR using inflammatory markers, facilitating the selection of the optimal treatment strategy. METHODS: Patients undergoing primary tumor resection after long-cycle NCRT at a single center (2012 to 2018) were retrospectively collected (n=130). Patient demographics, preoperative laboratory measurements, tumor characteristics, treatment strategy, and postoperative anatomopathological variables were collected. The association of factors to pCR was examined using binary logistic regression, odds ratio (OR) (95% confidence interval), and the discriminative capacity with the ROC curve. RESULTS: Out of 130 patients, 42 pCRs occurred, equal to 32.3% of the sample. Variables identified as useful to predict pCR were total neutrophil count (3; OR 7.6), intravenous 5-FU chemotherapy strategy (OR 3.2), and absence of diabetes (OR 3.4). Patients having all three of them had a 55.3% chance of pCR. CONCLUSIONS: The absolute neutrophil count better predicts pCR than other inflammatory indices in selected patients with LARC undergoing long-cycle NCRT. A neutrophil count less than 6400 cells/mm3, absence of diabetes, and intravenous 5-FU NCRT therapy lead to a relative rise in pCR