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

KNOWLEDGE, ATTITUDES AND PREFERENCES AMONG SPANISH COMMUNITY PHARMACISTS REGARDING INHALED THERAPY (THE OPTIM PHARMACY STUDY)

Objective: To assess knowledge, attitudes, and preferences regarding inhaled therapy among Spanish community pharmacists.Methods: An 11-item questionnaire was developed and distributed to community pharmacists throughout the country. Data collected included demographics, the source of knowledge of inhaler use, known and preferred devices, steps for correct use of metered-dose (pMDI) and dry-powder (DPI) inhalers, important variables when prescribing an inhaler device, patient education, and checking inhaler technique.Results: Of a total of 3000 questionnaires delivered, 1722 (57.4%) were returned. The most common source of knowledge was the package insert (46.9%) followed by personal experience (33.3%). DiskusTM and TurbuhalerTM were the best-known devices (96.4% and 93.4%), and DPIs the preferred inhalers. Although more than half of the surveyed pharmacists were aware of the most important step for correct inhalation with pMDI and DPI, only 18% identified the correct answer ‘Patient's preference' as the most important variable when prescribing an inhaler device. Most of the respondents had inadequate knowledge of inhaled therapies. Statistically, significant differences were found according to geographical areas. Moreover, the mean score on inhaled therapy with one knowledge source was higher than for those with none (P<0.05). Additionally, patient education was poor.Conclusion: In spite of the increasing involvement of Spanish community pharmacists in patients' care, their knowledge of inhaler use and attitudes towards inhaled therapy needs to improve, so that they can provide better patient education

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

Factor pronòstic; Carcinoma colorectal; Càrrega tumoralFactor pronóstico; Carcinoma colorrectal; Carga tumoralPrognostic factor; Colorectal Carcinoma; Tumor BurdenINTRODUCTION: 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, https://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 Investigación Científica y Técnica y de Innovación, and Agència de Gestió 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)

Lymph Node Molecular Analysis with OSNA Enables the Identification of pT1 CRC Patients at Risk of Recurrence: A Multicentre Study

OSNA; Lymph node; Colorectal cancerOSNA; Ganglio linfático; Cáncer colonrectalOSNA; Gangli limfàtic; Càncer colonrectalEarly-stage colorectal carcinoma (CRC)-pT1-is a therapeutic challenge and presents some histological features related to lymph node metastasis (LNM). A significant proportion of pT1 CRCs are treated surgically, resulting in a non-negligible surgical-associated mortality rate of 1.5-2%. Among these cases, approximately 6-16% exhibit LNM, but the impact on survival is unclear. Therefore, there is an unmet need to establish an objective and reliable lymph node (LN) staging method to optimise the therapeutic management of pT1 CRC patients and to avoid overtreating or undertreating them. In this multicentre study, 89 patients with pT1 CRC were included. All histological features associated with LNM were evaluated. LNs were assessed using two methods, One-Step Nucleic Acid Amplification (OSNA) and the conventional FFPE plus haematoxylin and eosin (H&E) staining. OSNA is an RT-PCR-based method for amplifying CK19 mRNA. Our aim was to assess the performance of OSNA and H&E in evaluating LNs to identify patients at risk of recurrence and to optimise their clinical management. We observed an 80.9% concordance in LN assessment using the two methods. In 9% of cases, LNs were found to be positive using H&E, and in 24.7% of cases, LNs were found to be positive using OSNA. The OSNA results are provided as the total tumour load (TTL), defined as the total tumour burden present in all the LNs of a surgical specimen. In CRC, a TTL ≥ 6000 CK19 m-RNA copies/µL is associated with poor prognosis. Three patients had TTL > 6000 copies/μL, which was associated with higher tumour budding. The discrepancies observed between the OSNA and H&E results were mostly attributed to tumour allocation bias. We concluded that LN assessment with OSNA enables the identification of pT1 CRC patients at some risk of recurrence and helps to optimise their clinical management.This research was supported by a grant from the Fondo de Investigación Sanitaria PI20/00863 awarded to M.C. and J.C., and PI19/01050 awarded to MP, and it was funded by Instituto de Salud Carlos III and Beca de la Marató de TV3 2020 (Beca la Marato—201932-30) awarded to MP. This research was also co-funded by the European Regional Development Fund/European Social Fund; “A way to make Europe”/“Investing in your future”. CIBEREHD is funded by the Instituto de Salud Carlos III

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)

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 H&E. 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