Evaluation of tumor infiltrating lymphocytes (TILs) according to the subtype of triple negative breast cancer and its association with the prognosis of the disease in Colombian patients

Objetivo: Relacionar el porcentaje y composición del infiltrado inmune del cáncer de mama triple negativo con el subtipo, y el pronóstico de la enfermedad en mujeres colombianas diagnosticadas en el Instituto Nacional de Cancerología y otras instituciones entre los años 2008 y 2016. Métodos: Se analizaron 195 tumores de cáncer de mama triple negativo (TN) de pacientes diagnosticadas entre 2008-2016 en 3 instituciones de salud de Colombia. Los linfocitos infiltrantes tumorales (TILs) estromales (sTILs) se evaluaron siguiendo las recomendaciones del International TILs Working Group 2014, en las láminas con tinción de hematoxilina y eosina de muestras pretratamiento. El número de células positivas para CD4, CD8 y PD-L1 se evaluó mediante técnicas de inmunohistoquímica (IHQ) y captura de imágenes digitales. Se analizo la expresión del receptor de andrógenos (RA) y la citoqueratina 5/6 (CK 5/6) por IHQ para la asignación de subtipos de TN. Se utilizaron pruebas paramétricas y no paramétricas para evaluar las diferencias en las características clínico-patológicas y el subtipo de cáncer de mama TN según los niveles y la composición de sTILs. Las diferencias en la supervivencia global (SG) y la supervivencia libre de enfermedad (SLE) se analizaron mediante curvas de Kaplan-Meier y la prueba de log-rank. Se utilizaron modelos multivariados para analizar la asociación de los sTILs y las poblaciones especificas con la SG, SLE y la respuesta patología completa (pCR). Resultados: Pacientes con altos niveles de sTILs presentaban con mayor frecuencia tumores en estadios tempranos (64.4% en estadios I/II) en comparación con aquellos con niveles bajos de sTILs (35.5%). Además, cuando se compararon pacientes con sTILs altos versus sTILs bajos, un mayor porcentaje de pacientes con sTILs altos no recibieron quimioterapia neoadyuvante (NAC) (alta: 50 % versus baja: 32.7 %, p=0.025) y recibieron cirugías más conservadoras (alta:60% vs. baja:37.9%, p=0.005). Se observaron resultados similares en pacientes con alta infiltración de células CD4/8 positivas. La positividad de PD-L1 fue más frecuentemente observada en pacientes con nacimiento o procedencia de la región andina y con obesidad. No se observaron diferencias estadísticamente significativas en el infiltrado inmune ni expresión de PD-L1 entre los subtipos de cáncer de mama TN. Se observaron tiempos de SG y SLE más prolongados en pacientes con sTILs elevados (p<0.001, p=0.022, respectivamente), así como en pacientes con infiltración elevada de CD4+ (p=0.007, p=0.021, respectivamente) y CD8+ (p=0.008, p=0.017, respectivamente). En el análisis multivariado, los niveles bajos de sTILs se encontraron como un factor pronóstico independiente asociado con un mayor riesgo de muerte (HR: 1.59, intervalo de confianza (IC) 95% 1.01 – 2.48). En cuanto a sTILs como biomarcador predictivo, un mayor porcentaje de pacientes con sTILs altos y células CD4+ altas lograron una respuesta clínica completa a NAC en comparación con los grupos de baja infiltración (sTILs: 28.6% vs 9.3%, p=0.022; CD4: 29% vs 9.4%, p=0.032) y pCR (sTILs: 42.9% vs. 15.8%, p=0.023; para CD4: 43.3% vs. 16.3%, p=0.006) en comparación con pacientes con baja infiltración. Asimismo, se observó una asociación estadísticamente significativa entre sTILs y pCR (OR: Alto sTILs 1.486, 95% IC 1.14 – 2.013). Se observaron resultados similares para la infiltración alta de CD4 y CD8 (OR: 1.262, IC 95 %: 1.061 – 1.536, OR: 1.337 IC 95 %: 1.085 – 1.694, respectivamente). Conclusiones: Nuestros resultados sugieren que los niveles de sTILs, CD4 y CD8 son un potencial marcador pronóstico de SG y un marcador predictivo de pCR en pacientes colombianas con cáncer de mama TN, sin embargo, es necesario continuar explorando la asociación del infiltrado inmune con el subtipo de cáncer de mama TN.Instituto Nacional de CancerologíaMincienciasObjective: To correlate the percentage and composition of the immune infiltrate of triple negative breast cancer with the subtype, and the prognosis of the disease in Colombian women diagnosed at the Instituto Nacional de Cancerología and other institutions between 2008 and 2016. Methods: A total of 195 triple negative (TN) breast cancer tumor biospecimens from patients diagnosed between 2008-2016 in 3 Colombian health institutions were included. Stromal TILs (sTILs) were evaluated following the recommendations of the International TILs Working Group 2014 in hematoxylin & eosin slides from pre-treatment samples. The number of positive cells for CD4, CD8 and PD-L1 was evaluated by immunohistochemistry and digital image capture. Parametric and non-parametric tests were used to evaluate differences in clinic-pathological characteristics and TN subtype according to sTILs levels and composition. Differences in overall survival (OS) and disease-free survival (DFS) were analyzed using Kaplan-Meier curves and the log-rank test. Cox regression analysis was used to analyze immune infiltrate as a prognostic marker for OS and DFS. A logistic regression model was applied to evaluate the association of immune infiltrate and pCR. Results: Tumors with high sTILs levels were more likely to be early stage (64.4% stage I/II) compared to tumors with low sTILs levels (35.5%). Additionally, when compared to patients with low sTILs, a higher percentage of patients with high sTILs didn’t receive neoadjuvant chemotherapy (NAC) (high:50% vs. low:32.7%, p=0.025) and received more conservative surgeries (high:60% vs. low:37.9%, p=0,005). Similar results were observed for patients with high CD4/8 infiltration levels. Positivity of PD-L1 were more likely occur in obese patients from Andean region. Longer OS and DFS times were observed in patients with high sTILs (p<0.001, p=0.022, respectively), as well as in patients with high CD4+ infiltration (p=0.007, p=0.021, respectively) and CD8+ infiltration (p=0.008, p=0.017, respectively). In the multivariate analysis, low levels of sTILs was found to be an independent prognostic factor associated with a higher risk of death (HR: 1.59, 95% CI 1.01-2.48). Regarding sTILs as a predictive biomarker, a higher number of patients with high sTILs and high CD4+ cells achieved clinical complete response to NAC (sTILs: 28.6% vs. 9.3%, p=0.022; CD4: 29% vs 9.4%, p=0.032) and pCR (sTILs: 42.9% vs. 15.8%, p=0.023; for CD4: 43.3% vs 16.3%, p=0.006) compared to patients with low infiltration. Likewise, a statistically significant association between sTILs and pCR was observed (OR: High sTILs 1.486, 95% CI 1.14 – 2.013). Similar results were observed for high CD4 and CD8 infiltration (OR: 1.262, 95% CI 1.061 – 1.536, OR: 1.337, 1.085 – 1.694, respectively). Conclusions: Our results suggest that sTILs, CD4 and CD8 levels are a prognostic marker for OS and a predictive marker for pCR in TN breast cancer patients from Colombian patients, however, it is necessary to continue exploring the association of the immune infiltrate with the subtype of TN breast cancer.Magíster en Ciencias BiológicasMaestríahttps://orcid.org/0000-0002-7962-8081https://scienti.minciencias.gov.co/cvlac/visualizador/generarCurriculoCv.do?cod_rh=000008728

Immune Lymphocyte Infiltrate and its Prognostic Value in Triple-Negative Breast Cancer

Triple-negative breast cancer (TNBC) occurs more frequently in young (&lt;50 years) non-Hispanic black and Hispanic/Latina women. It is considered the most aggressive subtype of breast cancer, although, recently, immune infiltrate has been associated with long-term survival, lower risk of death and recurrence, and response to neoadjuvant chemotherapy. The aim of this review was to evaluate the clinical impact of the immune infiltrate in TNBC by discussing whether its prognostic value varies across different populations. A comprehensive systematic search in databases such as PubMed and Web of Science was conducted to include papers focused on tumor-infiltrating lymphocytes (TILs) in TNBC in different population groups and that were published before January 2021. TNBC patients with higher levels of TILs had longer overall survival and disease-free survival times compared with TNBC patients with low TIL levels. Similar results were observed for CD4+, CD8+ TIL populations. On the other hand, patients with high TIL levels showed a higher rate of pathological complete response regardless of the population group (Asian, European, and American). These results altogether suggest that TIL subpopulations might have a prognostic role in TNBC, but the underlying mechanism needs to be elucidated. Although the prognosis value of TILs was not found different between the population groups analyzed in the revised literature, further studies including underrepresented populations with different genetic ancestries are still necessary to conclude in this regard

Evaluation of tumor infiltrating lymphocytes (TILs) according to the subtype of triple negative breast cancer and its association with the prognosis of the disease in Colombian patients

Objetivo: Relacionar el porcentaje y composición del infiltrado inmune del cáncer de mama triple negativo con el subtipo, y el pronóstico de la enfermedad en mujeres colombianas diagnosticadas en el Instituto Nacional de Cancerología y otras instituciones entre los años 2008 y 2016. Métodos: Se analizaron 195 tumores de cáncer de mama triple negativo (TN) de pacientes diagnosticadas entre 2008-2016 en 3 instituciones de salud de Colombia. Los linfocitos infiltrantes tumorales (TILs) estromales (sTILs) se evaluaron siguiendo las recomendaciones del International TILs Working Group 2014, en las láminas con tinción de hematoxilina y eosina de muestras pretratamiento. El número de células positivas para CD4, CD8 y PD-L1 se evaluó mediante técnicas de inmunohistoquímica (IHQ) y captura de imágenes digitales. Se analizo la expresión del receptor de andrógenos (RA) y la citoqueratina 5/6 (CK 5/6) por IHQ para la asignación de subtipos de TN. Se utilizaron pruebas paramétricas y no paramétricas para evaluar las diferencias en las características clínico-patológicas y el subtipo de cáncer de mama TN según los niveles y la composición de sTILs. Las diferencias en la supervivencia global (SG) y la supervivencia libre de enfermedad (SLE) se analizaron mediante curvas de Kaplan-Meier y la prueba de log-rank. Se utilizaron modelos multivariados para analizar la asociación de los sTILs y las poblaciones especificas con la SG, SLE y la respuesta patología completa (pCR). Resultados: Pacientes con altos niveles de sTILs presentaban con mayor frecuencia tumores en estadios tempranos (64.4% en estadios I/II) en comparación con aquellos con niveles bajos de sTILs (35.5%). Además, cuando se compararon pacientes con sTILs altos versus sTILs bajos, un mayor porcentaje de pacientes con sTILs altos no recibieron quimioterapia neoadyuvante (NAC) (alta: 50 % versus baja: 32.7 %, p=0.025) y recibieron cirugías más conservadoras (alta:60% vs. baja:37.9%, p=0.005). Se observaron resultados similares en pacientes con alta infiltración de células CD4/8 positivas. La positividad de PD-L1 fue más frecuentemente observada en pacientes con nacimiento o procedencia de la región andina y con obesidad. No se observaron diferencias estadísticamente significativas en el infiltrado inmune ni expresión de PD-L1 entre los subtipos de cáncer de mama TN. Se observaron tiempos de SG y SLE más prolongados en pacientes con sTILs elevados (p<0.001, p=0.022, respectivamente), así como en pacientes con infiltración elevada de CD4+ (p=0.007, p=0.021, respectivamente) y CD8+ (p=0.008, p=0.017, respectivamente). En el análisis multivariado, los niveles bajos de sTILs se encontraron como un factor pronóstico independiente asociado con un mayor riesgo de muerte (HR: 1.59, intervalo de confianza (IC) 95% 1.01 – 2.48). En cuanto a sTILs como biomarcador predictivo, un mayor porcentaje de pacientes con sTILs altos y células CD4+ altas lograron una respuesta clínica completa a NAC en comparación con los grupos de baja infiltración (sTILs: 28.6% vs 9.3%, p=0.022; CD4: 29% vs 9.4%, p=0.032) y pCR (sTILs: 42.9% vs. 15.8%, p=0.023; para CD4: 43.3% vs. 16.3%, p=0.006) en comparación con pacientes con baja infiltración. Asimismo, se observó una asociación estadísticamente significativa entre sTILs y pCR (OR: Alto sTILs 1.486, 95% IC 1.14 – 2.013). Se observaron resultados similares para la infiltración alta de CD4 y CD8 (OR: 1.262, IC 95 %: 1.061 – 1.536, OR: 1.337 IC 95 %: 1.085 – 1.694, respectivamente). Conclusiones: Nuestros resultados sugieren que los niveles de sTILs, CD4 y CD8 son un potencial marcador pronóstico de SG y un marcador predictivo de pCR en pacientes colombianas con cáncer de mama TN, sin embargo, es necesario continuar explorando la asociación del infiltrado inmune con el subtipo de cáncer de mama TN.Instituto Nacional de CancerologíaMincienciasObjective: To correlate the percentage and composition of the immune infiltrate of triple negative breast cancer with the subtype, and the prognosis of the disease in Colombian women diagnosed at the Instituto Nacional de Cancerología and other institutions between 2008 and 2016. Methods: A total of 195 triple negative (TN) breast cancer tumor biospecimens from patients diagnosed between 2008-2016 in 3 Colombian health institutions were included. Stromal TILs (sTILs) were evaluated following the recommendations of the International TILs Working Group 2014 in hematoxylin & eosin slides from pre-treatment samples. The number of positive cells for CD4, CD8 and PD-L1 was evaluated by immunohistochemistry and digital image capture. Parametric and non-parametric tests were used to evaluate differences in clinic-pathological characteristics and TN subtype according to sTILs levels and composition. Differences in overall survival (OS) and disease-free survival (DFS) were analyzed using Kaplan-Meier curves and the log-rank test. Cox regression analysis was used to analyze immune infiltrate as a prognostic marker for OS and DFS. A logistic regression model was applied to evaluate the association of immune infiltrate and pCR. Results: Tumors with high sTILs levels were more likely to be early stage (64.4% stage I/II) compared to tumors with low sTILs levels (35.5%). Additionally, when compared to patients with low sTILs, a higher percentage of patients with high sTILs didn’t receive neoadjuvant chemotherapy (NAC) (high:50% vs. low:32.7%, p=0.025) and received more conservative surgeries (high:60% vs. low:37.9%, p=0,005). Similar results were observed for patients with high CD4/8 infiltration levels. Positivity of PD-L1 were more likely occur in obese patients from Andean region. Longer OS and DFS times were observed in patients with high sTILs (p<0.001, p=0.022, respectively), as well as in patients with high CD4+ infiltration (p=0.007, p=0.021, respectively) and CD8+ infiltration (p=0.008, p=0.017, respectively). In the multivariate analysis, low levels of sTILs was found to be an independent prognostic factor associated with a higher risk of death (HR: 1.59, 95% CI 1.01-2.48). Regarding sTILs as a predictive biomarker, a higher number of patients with high sTILs and high CD4+ cells achieved clinical complete response to NAC (sTILs: 28.6% vs. 9.3%, p=0.022; CD4: 29% vs 9.4%, p=0.032) and pCR (sTILs: 42.9% vs. 15.8%, p=0.023; for CD4: 43.3% vs 16.3%, p=0.006) compared to patients with low infiltration. Likewise, a statistically significant association between sTILs and pCR was observed (OR: High sTILs 1.486, 95% CI 1.14 – 2.013). Similar results were observed for high CD4 and CD8 infiltration (OR: 1.262, 95% CI 1.061 – 1.536, OR: 1.337, 1.085 – 1.694, respectively). Conclusions: Our results suggest that sTILs, CD4 and CD8 levels are a prognostic marker for OS and a predictive marker for pCR in TN breast cancer patients from Colombian patients, however, it is necessary to continue exploring the association of the immune infiltrate with the subtype of TN breast cancer.Magíster en Ciencias BiológicasMaestríahttps://orcid.org/0000-0002-7962-8081https://scienti.minciencias.gov.co/cvlac/visualizador/generarCurriculoCv.do?cod_rh=000008728

Diagnóstico molecular una alternativa para la detección de patógenos en alimentos

Introduction: Foodborne diseases constitute a health problem in the world, involving products of animal origin, vegetables, water sources, and foods ready for consumption.  The problem arises because of the inadequate manipulation of food and the possibility of transmission of pathogenic microorganisms, mainly of a bacterial type.Objective: To describe the different conventional and molecular diagnostic techniques used in the food industry.Material and Methods: A search was carried though the analysis of review articles, original articles, and theses during a period of three months; these articles were written in English, Portuguese and Spanish.  In their content, they demonstrated the use of conventional and molecular methodology for the identification of microorganisms in different types of food.Results: The conventional methods for the identification of these microorganisms make use of well-established protocols and norms, but require too much time for their identification and, in some cases, they provide low sensitivity and specificity. Molecular techniques are proposed as an alternative for the microbiological evaluation of food because they are fast, sensitive and reliable.Conclusions: Molecular techniques are proposed as a new and faster alternative for the detection of microorganisms, reducing the time for the emission of results from days to hours; identifying pathogenesis factors, virulence and drug resistance with only one assemblage, thus providing high sensitivity and specificity.Keywords: Pathogen, Diseases, Food, Transmission, Molecular Diagnostic Techniques.Introducción: Las enfermedades transmitidas por alimentos (ETA) constituyen un problema de salud pública, en ellas se ven implicados productos de origen animal, vegetal, fuentes de agua y alimentos listos para consumo, debido a su manipulación y posibilidad de trasmisión de microorganismos patógenos principalmente de tipo bacteriano.Objetivo: Describir las diferentes técnicas de diagnóstico convencional y molecular empleadas en la industria alimentaria.Material y Métodos: Se realizó una búsqueda de artículos de revisión, originales y tesis durante un periodo de tres meses en inglés, portugués y español que en su contenido mostrarán el uso de metodología convencional y molecular para la identificación de microorganismos en alimentosResultados: Los métodos convencionales para la identificación de estos microorganismos hacen uso de protocolos y normas que están bien establecidos, pero requieren demasiado tiempo para su identificación y aportan baja sensibilidad y especificidad en algunos casos; las técnicas moleculares se plantean como una alternativa para la evaluación microbiológica de los alimentos, al ser rápidas, sensibles y fiables.Conclusiones: las técnicas moleculares se proponen como una nueva y más rápida alternativa de detección de microorganismos, disminuyen el tiempo para la emisión de los resultados de días a horas, identifican factores de patogenia, virulencia y resistencia a fármacos con tan solo un montaje, lo que aporta alta sensibilidad y especificidad.Palabras claves: patógeno, enfermedad, alimentos, transmisión, técnicas de diagnóstico molecular

Soluble HLA-G (sHLA-G) measurement might be useful as an early diagnostic biomarker and screening test for gastric cancer

Abstract Gastric cancer (GC) is the fifth most frequent malignancy worldwide and has a high mortality rate related to late diagnosis. Although the gold standard for the GC diagnosis is endoscopy with biopsy, nonetheless, it is not cost-effective and is invasive for the patient. The Human leukocyte antigen G (HLA-G) molecule is a checkpoint of the immune response. Its overexpression in cancer is associated with immune evasion, metastasis, poor prognosis, and lower overall survival. We evaluate the plasma levels of soluble HLA-G, (sHLA-G) in patients with GC and benign gastric pathologies using an ELISA test. A higher concentration of sHLA-G in patients with GC than in those with benign pathologies, higher levels of plasma sHLA-G in women with GC compared with men and significant differences in the sHLA-G levels between the benign gastric pathologies evaluated, was our main findings. As no significant differences were found between the GC assessed stages in our study population, we suggest that sHLA-G is not an adequate marker for staging GC, but it does have diagnostic potential. In addition to providing information on the potential of sHLA-G as a diagnostic marker for GC, our study demonstrate that HLA-G molecules can be found in the membrane of exosomes, which highlights the need to perform studies with a larger number of samples to explore the functional implications of HLA-G positive exosomes in the context of gastric cancer, and to determine the clinical significance and possible applications of these findings in the development of non-invasive diagnostic methods

Espirales de reflexividad crítica y propositiva para escribir la educación media de Bogotá

399 p. Libro digita