Deep Phenotypic Characterisation of CTCs by Combination of Microfluidic Isolation (IsoFlux) and Imaging Flow Cytometry (ImageStream)

Ines Aznar-Peralta holds a "Garantia Juvenil" fellowship (contract number 8040), and M. Carmen Garrido-Navas has a postdoctoral fellowship funded by the Ministry of Economy, Competitiveness, Enterprises and Universities (DOC_01682).The isolation of circulating tumour cells (CTCs) in colorectal cancer (CRC) mostly relies on the expression of epithelial markers such as EpCAM, and phenotypic characterisation is usually performed under fluorescence microscopy with only one or two additional markers. This limits the ability to detect different CTC subpopulations based on multiple markers. The aim of this work was to develop a novel protocol combining two platforms (IsoFluxTM and ImageStream®X) to improve CTC evaluation. Cancer cell lines and peripheral blood from healthy donors were used to evaluate the efficiency of each platform independently and in combination. Peripheral blood was extracted from 16 early CRC patients (before loco-regional surgery) to demonstrate the suitability of the protocol for CTC assessment. Additionally, peripheral blood was extracted from nine patients one month after surgery to validate the utility of our protocol for identifying CTC subpopulation changes over time. Results: Our protocol had a mean recovery efficiency of 69.5% and a limit of detection of at least four cells per millilitre. We developed an analysis method to reduce noise from magnetic beads used for CTC isolation. CTCs were isolated from CRC patients with a median of 37 CTCs (IQ 13.0–85.5) at baseline. CTCs from CRC patients were significantly (p < 0.0001) larger than cytokeratin (CK)-negative cells, and patients were stratified into two groups based on BRAFV600E and PD-L1 expression on CK-positive cells. The changes observed over time included not only the number of CTCs but also their distribution into four different subpopulations defined according to BRAFV600E and PD-L1 positivity. We developed a novel protocol for semi-automatic CTC isolation and phenotypic characterisation by combining two platforms. Assessment of CTCs from early CRC patients using our protocol allowed the identification of two clusters of patients with changing phenotypes over time."Garantia Juvenil" fellowship 8040Ministry of Economy, Competitiveness, Enterprises and Universities DOC_0168

AA-NAT, MT1 and MT2 Correlates with Cancer Stem-Like Cell Markers in Colorectal Cancer: Study of the Influence of Stage and p53 Status of Tumors

The characterization of colon cancer stem cells (CSCs) may help to develop novel diagnostic and therapeutic procedures. p53 loss increases the pool of CSCs in colorectal cancer (CRC). Recent reports suggest that the oncostatic effects of melatonin could be related to its ability to kill CSCs. Although there are no data linking the loss of p53 function and melatonin synthesis or signaling in cancer, melatonin does activate the p53 tumor-suppressor pathway in this disease. In this work, we analyze whether the expression of melatonin synthesis and signaling genes are related to the expression of CSC markers and the implication of p53 status in samples from patients with CRC. Arylalkylamine N-acetyltransferase (AA-NAT), MT1, and MT2 expression decreased in tumor samples versus normal mucosa samples in mutated p53 (mtp53) tumors versus those with wild-type p53 (wtp53). Further, AA-NAT and MT2 expression were lower in advanced stages of the disease in wtp53 tumors. On the contrary, CD44 and CD66c expression was higher in tumor versus normal mucosa in wtp53 tumors. Additionally, CD44 expression was higher in advanced stages of the disease regardless of the p53 status. Patients with CD44highCD66chigh and wtp53 tumors in advanced stages showed low expression of AA-NAT and MT2 in wtp53 tumors. These results could indicate a possible interaction of these pathways in CRC.This research was supported by grants from the Consejería de Salud de la Junta de Andalucía (PI-0677-2013). Josefa León acknowledges sponsorship from the Servicio Andaluz de Salud “Nicolás Monardes” program

Current surgical techniques in rectal cancer

El cáncer de recto es actualmente una de las enfermedades que implican una mayor morbimortalidad en la sociedad, tanto para la persona que lo sufre como para la esfera que lo rodea. La evolución en este tipo de tumores permiten abandonar el concepto clásico de grandes incisiones y extensas resecciones, para acercarnos al abordaje mínimamente invasivo, y a realizar la resección que sea únicamente necesaria, buscando preservar la integridad esfintérica en el paciente, y tratando de evitar en la medida de lo posible la confección de ostomías. La mayoría de las investigaciones que se realizan sobre el cáncer de recto, van encaminadas a la mejora en los tratamientos complementarios a la cirugía, como los tratamientos quimioterápicos neoadyuvantes, y los fármacos posteriores a la cirugía, es decir, la quimioterapia adyuvante. Únicamente un tercio de los estudios que se llevan a cabo son para mejorar la técnica quirúrgica. En este apartado, la aparición de la cirugía laparoscópica, el uso de dispositivos a través de orificios naturales para realizar cirugía local, la aparición de la cirugía combinada TaTME, y el auge de la cirugía robótica, parecen ser la antesala de lo que se desarrollará en un futuro respecto a las mejoras quirurgicas. En esta revisión, queremos poner de manifiesto un resumen del tratamiento quirúrgico del cáncer de recto, haciendo especial hincapié en las nuevas técnicas y avances, con una breve reseña histórica y anatómica, sin olvidar la importancia y seguridad para el paciente, tanto en la misma cirugía, como en los resultados oncológicos posteriores.Rectal cancer is currently one of the diseases that imply greater morbidity and mortality in society, both for people suffering and for the people around. The evolution in this type of tumors allows us to leave the classic concept of large incisions and extensive resections towards the minimally invasive approach, and to perform the resection that is only necessary, seeking to preserve the sphincteric integrity in the patient, and trying to avoid as much as possible ostomies. Most of rectal cancer research is aimed to improving complementary treatments to surgery, such as neoadjuvant chemotherapeutic treatments, and post-surgery drugs. Only one third of the studies carried out are focused on improvig the surgical technique. In this section, the appearance of laparoscopic surgery, the use of devices through natural holes to perform local surgery, the appearance of TaTME combined surgery, and the rise of robotic surgery, seem to be the prelude to what will be developed. in the future regarding surgical improvements. In this review, we want to highlight a summary of the surgical treatment of rectal cancer, with special emphasis on new techniques and advances and also a brief historical and anatomical review, also including the importance and safety for the patient both in the surgery and subsequent cancer follow-up

Endothelin-1 as a Mediator of Heme Oxygenase-1-Induced Stemness in Colorectal Cancer: Influence of p53

Heme oxygenase-1 (HO-1) is an antioxidant protein implicated in tumor progression, metastasis, and resistance to therapy. Elevated HO-1 expression is associated with stemness in several types of cancer, although this aspect has not yet been studied in colorectal cancer (CRC). Using an in vitro model, we demonstrated that HO-1 overexpression regulates stemness and resistance to 5-FU treatment, regardless of p53. In samples from CRC patients, HO-1 and endothelin converting enzyme-1 (ECE-1) expression correlated significantly, and p53 had no influence on this result. Carbon monoxide (CO) activated the ECE-1/endothelin-1 (ET-1) pathway, which could account for the protumoral effects of HO-1 in p53 wild-type cells, as demonstrated after treatment with bosentan (an antagonist of both ETRA and ETRB endothelin-1 receptors). Surprisingly, in cells with a non-active p53 or a mutated p53 with gain-of-function, ECE-1-produced ET-1 acted as a protective molecule, since treatment with bosentan led to increased efficiency for spheres formation and percentage of cancer stem cells (CSCs) markers. In these cells, HO-1 could activate or inactivate certain unknown routes that could induce these contrary responses after treatment with bosentan in our cell model. However more research is warranted to confirm these results. Patients carrying tumors with a high expression of both HO-1 and ECE-1 and a non-wild-type p53 should be considered for HO-1 based-therapies instead of ET-1 antagonists-based ones.Instituto de Salud Carlos IIIFEDER (PI18/01947)MINECO grant (DPI2017-84439-R)Nicolás Monardes Program from the Andalusian Health Service (C-0033-2015)FPU2019 fellowship (FPU19/02269) from the Ministerio de Ciencia, Innovación y Universidades (Spain

El compromís de l’ICM amb la igualtat de gènere en la recerca marina

2 pages, 1 figure[EN] Despite the drive towards equality between women and men in research institutions in recent years and the progress that has been made, there is still a long road ahead. Science is still associated with the male image, and women’s work and contributions are often invisible. Like many other spheres of society, research institutions contribute to the structural re(production) of gender inequalities. Women and men each tend to have more presence in certain scientific fields, with the horizontal segregation that this involves. Furthermore, the “glass ceiling” (the invisible barrier that hinders women’s access to the highest levels of decision-making or responsibility, limiting their professional careers) causes vertical segregation. Staying in research is an obstacle course in which there is a constant trickle of skills and talents, which is exacerbated for women. Moreover, research often seems blind to the gender dimension in its approach, content and analysis. […][ES] A pesar del impulso de la igualdad entre mujeres y hombres en las instituciones de investigación en los últimos años y de los avances logrados, aún queda un largo camino por recorrer. La ciencia constituye todavía un ámbito asociado a la imagen masculina, invisibilizando con frecuencia el trabajo y las aportaciones de las mujeres. Como muchas otras esferas de la sociedad, las instituciones de investigación contribuyen a la re(producción) estructural de les desigualdades de género. Mujeres y hombres tienden a concentrarse en determinados campos científicos con la segregación horizontal que ello implica. Por otra parte, el llamado “techo de cristal” –la barrera invisible que dificulta u obstaculiza el acceso de las mujeres a los niveles más altos de poder, de decisión o de responsabilidad, limitando sus carreras profesionales–, materializa la segregación vertical. La permanencia en la investigación es una carrera de obstáculos en la que se produce un goteo constante de capacidades y talentos, que se agrava para las mujeres. Además, la investigación parece a menudo ciega a la importancia de considerar la dimensión de género en su abordaje, contenido y análisis. […][CAT] Malgrat l’impuls envers la igualtat a les institucions de recerca dels darrers anys i els avenços assolits, encara hi ha un llarg camí per recórrer. La ciència constitueix encara un àmbit associat a la imatge masculina, invisibilitzant sovint el treball i les aportacions de les dones. Com moltes altres esferes de la societat, les institucions de recerca contribueixen a la re(producció) estructural de les desigualtats de gènere. Dones i homes tendeixen a concentrar-se en determinats camps científics amb la segregació horitzontal que això implica. Per altra banda, l’anomenat “sostre de vidre” –la barrera invisible que dificulta o obstaculitza l’accés de les dones als nivells més alts de poder, de decisió o de responsabilitat, encotillant les seves carreres professionals–, materialitza la segregació vertical. La permanència en la recerca és una carrera d’obstacles on es produeix un degoteig constant de capacitats i talents, agreujat entre les dones. A més, la recerca sembla sovint cega a la importància de considerar la dimensió de gènere en el seu abordatge, contingut i anàlisi. […]Peer reviewe

Population-based multicase-control study in common tumors in Spain (MCC-Spain): rationale and study design

Introduction: We present the protocol of a large population-based case-control study of 5 common tumors in Spain (MCC-Spain) that evaluates environmental exposures and genetic factors. Methods: Between 2008-2013, 10,183 persons aged 20-85 years were enrolled in 23 hospitals and primary care centres in 12 Spanish provinces including 1,115 cases of a new diagnosis of prostate cancer, 1,750 of breast cancer, 2,171 of colorectal cancer, 492 of gastro-oesophageal cancer, 554 cases of chronic lymphocytic leukaemia (CLL) and 4,101 population-based controls matched by frequency to cases by age, sex and region of residence. Participation rates ranged from 57% (stomach cancer) to 87% (CLL cases) and from 30% to 77% in controls. Participants completed a face-to-face computerized interview on sociodemographic factors, environmental exposures, occupation, medication, lifestyle, and personal and family medical history. In addition, participants completed a self-administered food-frequency questionnaire and telephone interviews. Blood samples were collected from 76% of participants while saliva samples were collected in CLL cases and participants refusing blood extractions. Clinical information was recorded for cases and paraffin blocks and/or fresh tumor samples are available in most collaborating hospitals. Genotyping was done through an exome array enriched with genetic markers in specific pathways. Multiple analyses are planned to assess the association of environmental, personal and genetic risk factors for each tumor and to identify pleiotropic effects. Discussion: This study, conducted within the Spanish Consortium for Biomedical Research in Epidemiology & Public Health (CIBERESP), is a unique initiative to evaluate etiological factors for common cancers and will promote cancer research and prevention in Spain.The study was partially funded by the “Accion Transversal del Cancer”, approved on the Spanish Ministry Council on the 11th October 2007, by the Instituto de Salud Carlos III-FEDER (PI08/1770, PI08/0533, PI08/1359, PS09/00773, PS09/01286, PS09/01903, PS09/02078, PS09/01662, PI11/01403, PI11/01889, PI11/00226, PI11/01810, PI11/02213, PI12/00488, PI12/00265, PI12/01270, PI12/00715, PI12/00150), by the Fundación Marqués de Valdecilla (API 10/09), by the ICGC International Cancer Genome Consortium CLL, by the Junta de Castilla y León (LE22A10-2), by the Consejería de Salud of the Junta de Andalucía (PI-0571), by the Conselleria de Sanitat of the Generalitat Valenciana (AP 061/10), by the Recercaixa (2010ACUP 00310), by the Regional Government of the Basque Country by European Commission grants FOOD-CT- 2006-036224-HIWATE, by the Spanish Association Against Cancer (AECC) Scientific Foundation, by the The Catalan Government DURSI grant 2009SGR1489

Participación de los receptores de Gastrina / Colecistokinina, y su interacción con la Melatonina en el crecimiento y diferenciación de células HT-29 de cáncer de colon

Tesis Univ. Granada. Departamento de Cirugía y sus Especialidades. Departamento de Fisiología. Leída en diciembre de 200

Prognostic differences between lymphoepithelioma-like colon carcinoma and colon adenocarcinoma.

Lymphoepithelioma-like colon carcinoma (LELC) is rare. The Epstein-Barr virus (EBV) hasn´t been implicated in the pathogenesis of LELC of the colon, but they may in fact be more strongly associated with MSI. Its treatment is identical to adenocarcinoma. However, lymphocyte infiltration and microsatellite instability have been associated with better prognosis

The interplay of contourite and mass-wasting recent sedimentary processes at the Guadalquivir Bank Margin uplift, Gulf of Cadiz: morphological high-resolution approach

European Geosciences Union General Assembly 2014 (EGU2014), 27 april - 2 may 2014, Vienna, Austria.-- 1 pageThe Gulf of Cadiz records the interplay of a variety of sedimentary processes related to the flow of the Mediterranean Outflow Water (MOW) exiting the Mediterranean Sea, with downslope sedimentary processes and the topography of the region. This work presents detailed morphological features of the Guadalquivir Ridge area, based on high resolution bathymetry and very-high resolution seismic profiles (TOPAS) acquired during the MONTERA cruise. The Guadalquivir Ridge is a SW-NE-oriented relief located on the middle slope of the Gulf of Cadiz (8º-7º10’ W). It reaches minimum depths at two highs, one at the Guadalquivir Bank, at the western extreme of the ridge (275 m), and a second one close to the eastern extreme (350 m). The ridge is cut by a gap where the Diego Cao contourite moat is incised forming a narrow, 4-5 km wide, SE-NW oriented channel. It delimits two contourite sheeted drifts (SD) at the northern side of the ridge: the Faro SD at the east ( 600 m water depth) and the Bartolomeo Dias SD, at the west (750 m water depth). The SD are relatively flat and become shallower progressively in a SE direction towards the Guadalquivir Ridge. At the SE side of the Guadalquivir Ridge depth increases dramatically where the Huelva and Cadiz contourite channels occur. They are originated by the direct erosion of the Lower Core of the MOW, running at depths of around 1200 m. The Diego Cao channel is related to the Upper Core, which runs at depths of around 800 m (Ambar and Serra, 2007). High resolution data reveal the existence of a variety of features. Semi-circular scarps, up to 10s km long, occur at the SE side of the Guadalquivir Ridge and at the SW side of the Bartolomeo Dias SD, at the rim of the Diego Cao contourite channel. Scarps occur at depths of 550 to 750 m and form steep steps of tens to hundreds of meters and in some cases occur overlapped one on each other at different depths. The second type of feature is a series of circular to ellipse-shaped depressions identified at the NE side of the Faro SD. Depressions are a few km in diameters and up to 100 m deep, and are aligned parallel to the edge of the SD, close to the rim of the Diego Cao. Finally, a valley-shaped depression has been identified at the N side of the Guadalquivir Bank. It is about 30 km long, with incision depths of up to 200 m and it runs parallel to the shape of the bank main relief. This work evaluates the relationship of the Lower and Upper cores of the MOW with the existing topography of the Guadalquivir ridge, as the origin for the identified morphologies, as the result of the interplay of mass-wasting and contouritic processesPeer Reviewe