Instituto de Oncología y Radiobiología

The CD6-CD318 axis has emerged as a potential target for immuno-oncology. Recent work has shown that blocking the CD6-CD318 interaction with a murine anti-human CD6 antibody increases lymphocyte cytotoxicity. However, several studies have demonstrated the drawbacks associated with the clinical use of murine antibodies and the variability among anti-CD6 antibodies. Therefore, evidence that the first-in-class humanized anti-human CD6 antibody itolizumab could be used for cancer immunotherapy may be a breakthrough in developing an antitumor clinical approach. Phenotypic and functional characterization of peripheral blood mononuclear cells (PBMCs) from healthy donors after challenge with CD318+ cell lines was performed by flow cytometry. In addition, IFNγ was determined by ELISA in culture supernatants. Immunohistochemical analyses of breast tumor samples were also performed. Here, we provide evidence supporting the rationale for itolizumab in cancer immunotherapy. The blockade of the CD6-CD318 interaction by itolizumab increases the cytotoxic capacity of CD8 T and NK cells over CD318+ tumor lines, reverses the NKG2A/NKG2D ratio, and increases granzyme B and IFNγ production. Itolizumab also regulates immune responses by downregulating CD5 expression and upregulating PD-1 and CTLA-4 inhibitory receptors on lymphocytes, which contribute to reducing exacerbated responses and additively enhancing CD318+ tumor cell cytotoxicity when combined with other immunocheckpoint inhibitors. In addition, we report that CD6-CD318 interaction inhibits lymphocyte proliferation and survival while downregulating CD6 expression on lymphocytes in vitro and in human breast cancer tissue samples, reinforcing the role of the CD6-CD318 axis as an immune checkpoint and highlighting the potential of itolizumab as an immune checkpoint inhibitor. Taken together, our results provide the first evidence linking the blocking of the CD6-CD318 axis by itolizumab with the potentiation of functional properties of lymphocytes, highlighting itolizumab as a novel promising immunotherapy for CD318+ tumors and supporting the relevance of new combinatorial therapies with checkpoint inhibitors.

Background: Dermatophytosis are the leading cause of fungal infections worldwide. Objective: To compare the efficacy and safety of ozonated sunflower oil versus ketoconazole and terbinafine in patients with dermatophytosis. Methods: Multicenter Phase III study, it had an open, randomized, comparative design, with three parallel group to which ozonated sunflower oil was apply, or ketoconazole, or terbinafine, twice a day for 6 weeks. The effect on the clinical and mycological cure rate was prefix as the primary efficacy variable. The effect on treatment time to achieve cure was consider as a secondary efficacy variable. All data were analyzed by the intention-to-treat method. Results: The study included 300 patients of both sexes with a diagnosis of dermatophytosis, who were randomly distributed into three groups that received ozonated sunflower oil, or ketoconazole, or terbinafine, respectively. Treatment with ozonated sunflower oil produced a total clinical and mycological cure rate in 78% of patients, similar to ketoconazole (78%) and terbinafine (77%) treatment. Complete mycological cure was achieved in 87% of patients treated with ozonated sunflower oil, similar to ketoconazole (88%) and terbinafine (89%) treatments. Regarding the healing time, of the 233 patients with complete clinical cure, 10 patients (4.3%) were completely cure after 2 weeks of treatment, 58 patients (24.9%) after 4 weeks and 164 patients (70.4%) after 6 weeks. The treatments were safe and well tolerated.

Introduction: Chronic kidney disease (CKD) affects approximately 10% of the Spanish population and constitutes an independent cardiovascular risk factor. Vascular calcifications, especially in the abdominal aorta, are significantly associated with increased cardiovascular mortality in patients with CKD. This systematic review evaluates the evidence on the usefulness of ultrasound to detect and assess vascular calcifications in patients with CKD, and its relevance as a cardiovascular risk factor.Methods: A systematic search of multiple electronic databases was conducted until July 2024. Original studies published between 2000 and 2024 evaluating the use of ultrasound to detect vascular calcifications in adult patients with CKD at any stage were included. Reviews, letters, editorials, animal studies and those not published in English or Spanish were excluded. Methodological quality was assessed using the Newcastle-Ottawa scale for observational studies and the Cochrane tool for clinical trials.Results: Twenty-eight studies were included with a total of 50 to 3,000 participants per study. Most were observational studies, with 3 randomised clinical trials. Ultrasound showed good correlation with computed tomography (r=0.65-0.82) to quantify aortic and femoral calcifications. The presence of aortic calcifications was associated with increased risk of cardiovascular mortality (HR 1.8-3.2) in patients with advanced CKD and on haemodialysis. Iliac/femoral calcifications were associated with increased risk of cardiovascular events (RR 1.6-2.4) in patients with stage 3-5 CKD. Incorporation of ultrasound assessment of calcifications into CKD-specific risk algorithms significantly improved their predictive performance.Discussion: Ultrasonography emerges as a valuable tool for the detection of vascular calcifications due to its non-invasive nature, low cost and ability to perform repeated assessments. However, limitations such as heterogeneity in assessment methods, variable sample sizes and lack of long-term follow-up in some studies were identified. Further research is needed to standardise protocols and assess the long-term impact of interventions based on ultrasound detection of calcifications.Conclusions: Ultrasonography is an effective and accessible tool for detecting and assessing vascular calcifications in patients with CKD, providing relevant prognostic information for cardiovascular risk stratification. Its consideration as a first-line tool in nephrological practice is recommended. However, large-scale prospective studies are needed to definitively validate its prognostic value in different stages of CKD and to evaluate the efficacy of early interventions based on these findings.

Context: Marine ecosystems are sources of bioactive compounds. Thirty-eight microorganism strains from the Cuban platform were screened, which allowed us to identify an extract from Bacillus amyloliquefaciens, strain CBM-116, as a source for obtaining bioproducts with biomedical applications. Aims: To physiologically characterize the culture of Bacillus amyloliquefaciens (CBM-116 strain) and to evaluate the antioxidant and toxic potentialities in vitro of the M116 extract obtained from CBM-116. Methods: The growth and metabolite production of the culture were evaluated at a sieve scale. The chemical composition of the M116 extract obtained from the fermented CBM-116 culture was qualitatively characterized. The extract antioxidant activity was measured by DPPH• and FRAP assays, while cytotoxicity was evaluated in MDCK, J774, CT26, 4T1, MCF-7, A549 cell lines and in Caulobacter crescentus, as well as the effects on genetic material by SOS colorimetric and Rifampicin Resistance, in the last model. Results: Grow kinetic parameters of CBM-116 showed the formation of protein metabolites, while the extract revealed antioxidant capacity, which was evidenced by its iron-reducing capacity. M116 was not cytotoxic up to 2000 μg/mL in C. crescentus; however, it induced mutagenicity and primary damage to the DNA of the bacteria. The extract significantly inhibited cell viability of CT26, 4T1, MCF-7, A549 cells after 48 hours’ exposure. Mean inhibitory concentration (IC50) was calculated for CT26 and 4T1 cells with values of 384 and 488 µg/mL, respectively, in the MTT assay. In the neutral red assay, the values were 478.6 and 398 µg/mL, respectively. Meanwhile, the selectivity index showed values above 2 for both assays. MDCK and J774 cells were not affected. Conclusions: The M116 extract obtained from B. amyloliquefaciens showed bioactive properties with potential application for developing new anti-tumor agents.

Abstract Cancer is the second leading cause of death worldwide. Peptidases participate in tumor development and growth. Mammalian neutral aminopeptidase (APN, EC 3.4.11.2, M1 family) catalyzes the cleavage of neutral and basic amino acids from the N-terminus of substrates. APN expression is dysregulated in several types of cancer, being a target for the development of new anticancer agents. Recently, we identified three new non-competitive inhibitors of soluble porcine APN (pAPN) by virtual screening (BTB11079, JFD00064, BTB07018, from Maybridge). In the present contribution we assayed their effect on the activity of APN in a microsomal preparation of porcine kidney cortex, a model of the physicochemical environment of the enzyme. These classical inhibitors had an IC50 value of 3–5 µM. Additionally, using a kinetic approach and a specific substrate, we quantified APN activity on the cell surface of human and murine lung, colon, prostate, and skin tumor cells. APN inhibitors reduced tumor cells viability, more efficiently in the higher APN activity tumor cell lines, but not in non-tumoral cells. BTB11079, JFD00064, BTB07018 effects on cell viability were stronger than that of bestatin, a positive control. Thus, these non-competitive APN inhibitors may be useful tools for cancer treatment.

Background:Oral squamous cell carcinoma (OSCC) is the most common malignancy in this region, and thus, further elucidation of its tumoral mechanisms is important. One of the main roles of the acute-phase protein orosomucoid-1 (ORM1) is the promotion of angiogenesis, which is key for tumor nutrition and growth.Aim:Our aim was to evaluate the immunohistochemical expression of ORM1 and the angiogenic activity indicated by microvascular density (MVD) in OSCC samples according to histological grade.Materials and Methods:Formalin-fixed, paraffin-embedded sections from 45 OSCC cases were submitted to immunohistochemistry: 25 were well-differentiated OSCC, 18 were moderately differentiated OSCC and 2 were poorly differentiated OSCC. ORM1 staining was evaluated by a semiquantitative method, and CD34-positive blood vessels were quantified to calculate the MVD. The results were statically analyzed.Results:All cases exhibited immunoexpression of ORM1 and CD34. However, no significant differences were found between the expression of both markers among the histological grades. In addition, the presence of ORM1 in inflammatory cells and in the extracellular matrix was detected in most cases.Conclusion:These results suggest that the induction of angiogenesis is not the main role of ORM1 in OSCC and may be associated with the regulation of the immune/inflammatory response or the transport of protumoral molecules, such as sialyl-Lewis X or phorbol esters, which requires confirmation in future studies.

Differentiated thyroid carcinoma (DTC) incidence is characterized by wide ethnic and geographic variations, with high incidence rates observed in Oceanian populations. Genome-wide association studies (GWAS) identified mainly four DTC susceptibility loci at 9q22.33, 14q13.3, 2q35 and 8p12. Here we performed fine-mapping of the 2q35 and 8p12 loci in the population of the EPITHYR consortium that includes Europeans, Melanesians and Polynesians to identify likely causal variants for DTC risk. We conducted a colocalization analysis using eQTLs data to determine the SNPs with the highest probability of causality.At 2q35, we highlighted rs16857609 located in DIRC3. This SNP has a high probability of causality in the three populations, and a significant association in Europeans (OR = 1.4, p = 1.9 x 10-10). It is also associated with expression of DIRC3 and of the nearby gene IGFBP5 in thyroid tumour cells. At 8p12, we identified rs7844425 which was significantly associated with DTC in Europeans (OR = 1.32, p = 7.6 x 10-8) and rs2439304, which was highlighted by the colocalization analysis but only moderately associated with DTC in our dataset (OR = 1.2, p = 0.001). These SNPs are linked to the expression of NRG1 in thyroid tissue.Hence, our study identified novel variants at 2q35 and 8p12 to be prioritized for further functional studies.

En América Latina, las anomalías congénitas, después de la prematuridad, son la segunda causa de muerte de recién nacidos y niños menores de 5 años. En Ecuador, las malformaciones congénitas mayores ocuparon la cuarta causa de mortalidad infantil en 2014. Las mismas pueden afectar gravemente a los enfermos, sus familias y la sociedad. En ocasiones es complejo identificar las causas que las generan. Por tal motivo, el objetivo de esta investigación fue demostrar el aporte de la intervención comunitaria en la detección de factores de riesgo relacionados con malformaciones congénitas en recién nacidos en el Hospital Dr. Verdi Cevallos Balda (Portoviejo, Ecuador) en el período de enero de 2018 a junio de 2019. Se realizó un estudio descriptivo, retrospectivo y transversal. La muestra estuvo constituida por 56 nacidos vivos que presentaron anomalías congénitas. Se determinó que los factores de riesgo más frecuentemente asociados a malformaciones congénitas fueron los infecciosos durante la gestación, edad materna mayor a 35 años, anemia y desnutrición de la madre, y obesidad; además del consumo de tabaco y presencia de un hijo anterior con anomalías. La mortalidad neonatal se asoció directamente a las anomalías congénitas estructurales.

<p>La dipeptidil peptidasa IV (DPP-IV, EC 3.4.14.5), también conocida como CD26, es una aminopeptidasa de tipo serino con preferencia de corte por la secuencia Xaa-Pro o Xaa-Ala del extremo amino de péptidos provocando su activación/inactivación. La enzima es un homodímero y cada subunidad consiste en dos dominios: un dominio αβ-hidrolasa y un dominio de propela-β, ambos implicados en su función enzimática y su interacción con otras proteí-nas. Esta enzima desempeña funciones importantes en múltiples procesos fisiológicos relacionados con el meta-bolismo de la glucosa, por lo que constituye hoy en día uno de los blancos para el tratamiento de la diabetes me-llitus tipo 2. Esta enzima se encuentra también sobre-expresada en la artritis reumatoidea, la psoriasis, la colitis, la esclerosis múltiple y el rechazo a los trasplantes. DPP-IV participa además, en la regulación de la respuesta in-mune mediada por linfocitos CD4+. Recientemente, se ha observado que su actividad se altera en diferentes tipos de cáncer tales como: tiroides, ovario, pulmón, piel, próstata, tumores del sistema nervioso central, entre otros. En la clínica, los inhibidores de DPP-IV tienen efectos beneficiosos como anti-hiperglicémicos, protectores de los islotes pancreáticos, reguladores del sistema inmune, protectores de las funciones cardiovasculares y renales, y efectos anticancerígenos y anti-inflamatorios. Por estas razones, esta enzima constituye actualmente un impor-tante foco de atención para el diagnóstico, al ser considerada como un potencial marcador molecular de determi-nados tipos de patologías, así como para el desarrollo de inhibidores para combatir la diabetes mellitus tipo 2 y el cáncer. Los organismos marinos y costeros son una fuente abundante de compuestos bioactivos que incluyen pep-tidasas e inhibidores de peptidasas de todas las clases mecanísticas, siendo los inhibidores de tipo serino los más estudiados. En la presente contribución centramos la atención en las diferentes estrategias desarrolladas para la identificación y caracterización de inhibidores de DPP-IV a partir de organismos marinos y costeros. La misma de-muestra que la biodiversidad marina es una fuente importante, promisoria y aun poco explorada de inhibidores de DPP-IV con potenciales aplicaciones biomédicas en enfermedades humanas.</p>