Biomarkers of basal cell carcinoma resistance to methyl-aminolevulinate photodynamic therapy

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Background Methyl-aminolevulinate photodynamic therapy (MAL-PDT) is an excellent option for the treatment of basal cell carcinoma (BCC). However, up to 25% of cases are resistant to this treatment modality. Objective The aim of this study was to identify potential biomarkers of BCC response to MAL-PDT. Material and methods Clinical, histological, and immunohistochemical (p53, Ki-67, CD-31, COX2, β-catenin, EGFR, and survivin) variables were analyzed in a retrospective study of consecutive BCC patients treated with MAL-PDT at the San Jorge Hospital, Huesca, Spain between January 2006 and December 2015. To deepen on these markers, the effects on p53 and cyclin D1 expression, in vitro response to MAL-PDT of 2 murine BCC cell lines (ASZ and BSZ), was also evaluated. Results The retrospective study examined the response to MAL-PDT of 390 BCCs from 182 patients. The overall clinical response rate was 82.8%, with a mean follow-up time of 35.96 months (SD = 23.46). Immunohistochemistry revealed positive p53 in 84.6% of responders but only 15.4% of nonresponsive tumors (p = 0.011). Tumors with increased peripheral palisading of basal cell islands to immunostaining β-catenin responded poorly to PDT (p = 0.01). In line with our findings in patients, in vitro studies revealed a better response to PDT in the p53-positive ASZ cell line than the p53-negative BSZ cell line (p<0.01). Multivariate analysis revealed that the following variables were significantly associated with response to PDT: age, nBCC, presence of peritumoral inflammatory infiltrate, and p53 immunopositivity. Patients with positive p53 immunostaining were 68.54 times more likely to achieve cure than p53-negative patients (CI95% 2.94–159.8) Conclusion Our finding suggest that certain clinicopathological and immunohistochemical variables, particularly p53 expression, may serve as indicators of BCC response to MAL-PDT, and thus facilitate the selection of patients who are most likely to benefit from this therapyThis project received support from the Instituto de Salud Carlos III and Fondos Feder Europeos, MINECO (FIS PI15/00974). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscrip

Dual Role of Subphthalocyanine Dyes for Optical Imaging and Therapy of Cancer

This is the peer-reviewed version of the following article: van de Winckel, E., Mascaraque, M., Zamarrón, A., Juarranz de la Fuente, Á., Torres, T., & de la Escosura, A. (2018). Dual role of subphthalocyanine dyes for optical imaging and therapy of cancer. Advanced Functional Materials, 28(24), 1705938., which has been published in final form at https://doi.org/10.1002/adfm.201705938. This article may be used for non-commercial purposes in accordance with Wiley-VCH Terms and Conditions for Self-ArchivingThe family of subphthalocyanine (SubPc) macrocycles represents an interesting class of nonplanar aromatic dyes with promising features for energy conversion and optoelectronics. The use of SubPcs in biomedical research is, on the contrary, clearly underexplored, despite their documented high fluorescence and singlet oxygen quantum yields. Herein, for the first time it is shown that the interaction of these chromophores with light can also be useful for theranostic applications, which in the case of SubPcs comprise optical imaging and photodynamic therapy (PDT). In particular, the article evaluates, through a complete in vitro study, the dual-role capacity of a novel series of SubPcs as fluorescent probes and PDT agents, where the macrocycle axial substitution determines their biological activity. The 2D and 3D imaging of various cancer cell lines (i.e., HeLa, SCC-13, and A431) has revealed, for example, different subcellular localization of the studied photosensitizers (PS), depending on the axial substituent they bear. These results also show excellent photocytotoxicities, which are affected by the PS localization. With the best dual-role PS, preliminary in vivo studies have demonstrated their therapeutic potential. Overall, the present paper sets the bases for an unprecedented biomedical use of these well-known optoelectronic materials.E.v.d.W. and M.M. contributed equally to this work. The research leading to these results has received funding from the People Programme (Marie Curie Actions) of the European Union's Seventh Framework Program FP7‐PEOPLE‐2012‐ITN under REA grant agreement No. GA 316975. AdlE holds a Ramón y Cajal contract from the Spanish Ministry of Economy (MINECO). This work was supported by EU (CosmoPHOS‐nano, FP7‐NMP‐2012‐6, 310337‐2; GLOBASOL, FP7‐ENERGY‐2012‐J, 309194‐2), the Spanish MINECO (CTQ‐2014‐52869‐P (TT) and CTQ‐2014‐53673‐P (AdlE)), CAM (FOTOCARBON, S2013/MIT‐2841), grants from Instituto de Salud Carlos III, MINECO and Feder Funds (PI15/00974) and by S2010/BMD‐2359 from Comunidad de Madrid

Influence of Serum Vitamin D Level in the Response of Actinic Keratosis to Photodynamic Therapy with Methylaminolevulinate

In mouse models of squamous cell carcinoma, pre-treatment with calcitriol prior to photodynamic therapy with aminolevulinic acid (ALA) enhances tumor cell death. We have evaluated the association between vitamin D status and the response of actinic keratoses to photodynamic therapy with methylaminolevulinate. Twenty-five patients with actinic keratoses on the head received one session of photodynamic therapy with methylaminolevulinate. Biopsies were taken at baseline and six weeks after treatment. Immuno-histochemical staining was performed for VDR, P53, Ki67 and beta-catenin. Basal serum 25(OH)D levels were determined. Cases with a positive histological response to treatment had significantly higher serum 25(OH)D levels (26.96 (SD 7.49) ngr/mL) than those without response (18.60 (SE 7.49) ngr/mL) (p = 0.05). Patients with a complete clinical response displayed lower basal VDR expression (35.71% (SD 19.88)) than partial responders (62.78% (SD 16.735)), (p = 0.002). Our results support a relationship between vitamin D status and the response of actinic keratoses to photodynamic therapy with methylaminolevulinate

Nano- and microstructural evolution of alginate beads in simulated gastrointestinal fluids. Impact of M/G ratio, molecular weight and pH

peer-reviewedAlginate microcapsules were prepared using three different alginate grades and incubated under simulated digestion conditions. Their micro- and nanostructural changes were studied using microscopy, laser diffraction and small angle X-ray scattering. Both the molecular weight and M/G ratio affected the size and nanostructural features of the capsules, but the changes in gastrointestinal conditions were mainly determined by the latter. All microcapsules swelled slightly in simulated gastric fluid (pH = 3) and swelled further in simulated intestinal fluid (pH = 7), particularly those with high mannuronic acid (M) contents. While high guluronic acid (G) beads maintained the nanostructural features characteristic of alginate gels (junction zones) in both media, these were rapidly disrupted in the M-rich capsules. Decreasing the pH of the gastric phase from 3 to 2 had dramatic structural impacts, resulting in a greater integrity of the microcapsules, thus highlighting the importance of the selected digestion protocol for rational microcapsule design

Papel del tejido adiposo en las propiedades de las células madre tumorales del cáncer de páncreas

El adenocarcinoma ductal pancreático (ADP) es uno de los tumores más mortales con peor pronóstico, cuya incidencia se prevé que aumente en la próxima década. La obesidad constituye un factor de riesgo importante que puede influir promoviendo la progresión tumoral. Por otro lado, su agresividad intrínseca, potencial metastásico y quimioresistencia, se debe, en gran medida, a la subpoblación de células madre tumorales (CSCs) que contiene. Las CSCs pancreáticas son altamente dependientes del metabolismo lipídico, lo que favorece el fenotipo de CSC. En particular, los triacilglicéridos (TAGs) almacenados en gotas lipídicas constituyen una fuente principal energética para las células cancerosas. Estudios previos han asociado la captación de lípidos del microambiente tumoral con la progresión tumoral del ADP. Por ello, el objetivo de este trabajo es estudiar la comunicación entre los adipocitos y las CSCs en cáncer de páncreas.En el presente estudio se ha determinado por análisis bioinformático una sobre-expresión de genes del metabolismo lipídico en el tejido de ADP. La incubación de cultivos primarios de ADP con medio condicionado de adipocitos redujo el contenido de CD133+ e incrementó en esta subpoblación el contenido lipídico. Como consecuencia, se optó por un cocultivo indirecto entre esferas de ADP y adipocitos. Por un lado, el contenido de TAGs en el sobrenadante de cocultivos incrementó, mientras que en los adipocitos en cocultivo hubo una menor acumulación de TAGs. Además, en las esferas de ADP hubo un ligero incremento en el contenido lipídico de las CD133+. Por otro lado, no hubo diferencias en el contenido de CD133+ de las esferas provenientes de cocultivo, confirmado por la expresión de los genes de pluripotencia. Por último, los genes de EMT, SLUG y ZEB1 se sobre-expresaron. Además, se determinó que los lípidos derivados de los adipocitos podrían ser captados por el transportador CD36, sobre-expresado en esferas de ADP.En conclusión, los adipocitos desempeñan un papel importante sobre la funcionalidad de las CSCs pancreáticas, así como induciendo cambios metabólicos en las mismas. De hecho, los adipocitos actúan como donadores de lípidos en las células de ADP, pero el destino de los mismos no está claro ya que existen indicios de un comportamiento diferencial entre líneas celulares. Como resultado, esta puesta a punto ha demostrado que existe una comunicación entre los adipocitos y las CSCs de ADP, lo que incita a profundizar más en el papel del metabolismo lipídico y concretamente del tejido adiposo en el ADP.<br /

Tuning the nanoaggregates of sialylated biohybrid photosensitizers for intracellular activation of the photodynamic response

In the endeavor of extending the clinical use of photodynamic therapy (PDT) for the treatment of superficial cancers and other neoplastic diseases, deeper knowledge and control of the subcellular processes that determine the response of photosensitizers (PS) are needed. Recent strategies in this direction involve the use of activatable and nanostructured PS. Here, both capacities have been tuned in two dendritic zinc(II) phthalocyanine (ZnPc) derivatives, either asymmetrically or symmetrically substituted with 3 and 12 copies of the carbohydrate sialic acid (SA), respectively. Interestingly, the amphiphilic ZnPc-SA biohybrid (1) self-assembles into well-defined nanoaggregates in aqueous solution, facilitating cellular internalization and transport whereas the PS remains inactive. Within the cells, these nanostructured hybrids localize in the lysosomes, as usually happens for anionic and hydrophilic aggregated PS. Yet, in contrast to most of them (e. g., compound 2), hybrid 1 recovers the capacity for photoinduced ROS generation within the target organelles due to its amphiphilic character; this allows disruption of aggregation when the compound is inserted into the lysosomal membrane, with the concomitant highly efficient PDT responseThis work was supported by EU (CosmoPHOS-nano, EU-FP7- NMP-2012-LARGE-6, 310337), MINECO-Feder funds (CTQ2017- 85393-P (T.T.), CTQ-2014-53673-P and CTQ-2017-89539-P (A.d.l.E.), Instituto de Salud Carlos III; PI18/00708 (A.J.), and PCIN-2017-042/EuroNanoMed2017-191, TEMPEAT (T.T.)), and Comunidad Autonoma de Madrid (FOTOCARBON, S2013/MIT 2841). DLS measurements were carried out by VAM with a Nanotrac Wave analyzer, during an internship in the MESA+ Institute for Nanotechnology of the University of Twente. IMDEA Nanociencia also acknowledges support from the “Severo Ochoa” Programme for Centres of Excellence in R&D (MINECO, grant SEV-2016-0686

Protective effect of the aqueous extract of Deschampsia antarctica (EDAFENCE®) on skin cells against blue light emitted from digital devices

Skin is being increasingly exposed to artificial blue light due to the extensive use of electronic devices. This, together with recent observations reporting that blue light—also known as high-energy visible light—can exert cytotoxic effects associated with oxidative stress and promote hyperpigmentation, has sparked interest in blue light and its potential harmful effects on skin. The photoprotective properties of new extracts of different botanicals with antioxidant activity are therefore being studied. Deschampsia antarctica (Edafence®, EDA), a natural aqueous extract, has shown keratinocyte and fibroblast cell protection effects against ultraviolet radiation and dioxin toxicity. In this regard, we studied the protective capacity of EDA against the deleterious effects of artificial blue light irradiation in human dermal fibroblasts (HDF) and melanocytes. We analyzed the impact of EDA on viability, cell morphology, oxidative stress, melanogenic signaling pathway activation and hyperpigmentation in HDF and melanocytes subjected to artificial blue light irradiation. Our results show that EDA protects against cell damage caused by artificial blue light, decreasing oxidative stress, melanogenic signaling pathway activation and hyperpigmentation caused by blue light irradiation. All these findings suggest that EDA might help prevent skin damage produced by artificial blue light exposure from screen of electronic devicesThis research was funded by Cantabria Labs and by the Spanish grant from Instituto de Salud Carlos III, MINECO and FEDER funds (PI18/00708). M.G. and P.D.-W are supported from the Spanish Ministry (MINECO) and from Comunidad Autónoma de Madrid (CAM), respectively. The APC was funded by Cantabria Lab

Assessing amphiphilic ABAB Zn(II) phthalocyanines with enhanced photosensitization abilities in in vitro photodynamic therapy studies against cancer

We have previously demonstrated that singlet oxygen photosensitization abilities of Zn(II) phthalocyanines (Zn(II)Pcs) are enhanced through α-functionalization with bulky fluorinated substituents (i.e., bis(trifluoromethyl)phenyl units) at facing positions of ABAB Zn(II)Pcs, where A and B refer to differently functionalized isoindoles. In this work, we have prepared the Zn(II)Pc ABAB 1 endowed with hydrophilic triethylene glycol monomethyl ether (i.e., at the A isoindoles) to provide solubility in aqueous media, together with its A3B and A4 counterparts, and compared their ability to behave as photosensitizers for photodynamic therapy. All photophysical data, aggregation studies and preliminary in vitro biological assays in cell cultures of SCC-13 (squamous cell carcinoma) and HeLa (cervical cancer cells), have proved ABAB 1 as the best photosensitizer of the seriesThis research was funded by MINECO, Spain (CTQ2017-85393-P and CTQ2016-78454-C2-1-R) , Instituto de Salud Carlos III and Feder Funds (FIS PI18/00708) and ERANET/MINECO EuroNanoMe

Metformin overcomes metabolic reprogramming-induced resistance of skin squamous cell carcinoma to photodynamic therapy

Cancer metabolic reprogramming promotes resistance to therapies. In this study, we addressed the role of the Warburg effect in the resistance to photodynamic therapy (PDT) in skin squamous cell carcinoma (sSCC). Furthermore, we assessed the effect of metformin treatment, an antidiabetic type II drug that modulates metabolism, as adjuvant to PDT. Methods: For that, we have used two human SCC cell lines: SCC13 and A431, called parental (P) and from these cell lines we have generated the corresponding PDT resistant cells (10GT). Results: Here, we show that 10GT cells induced metabolic reprogramming to an enhanced aerobic glycolysis and reduced activity of oxidative phosphorylation, which could influence the response to PDT. This result was also confirmed in P and 10GT SCC13 tumors developed in mice. The treatment with metformin caused a reduction in aerobic glycolysis and an increase in oxidative phosphorylation in 10GT sSCC cells. Finally, the combination of metformin with PDT improved the cytotoxic effects on P and 10GT cells. The combined treatment induced an increase in the protoporphyrin IX production, in the reactive oxygen species generation and in the AMPK expression and produced the inhibition of AKT/mTOR pathway. The greater efficacy of combined treatments was also seen in vivo, in xenografts of P and 10GT SCC13 cells. Conclusions: Altogether, our results reveal that PDT resistance implies, at least partially, a metabolic reprogramming towards aerobic glycolysis that is prevented by metformin treatment. Therefore, metformin may constitute an excellent adjuvant for PDT in sSCCThis research was supported by Spanish grants from Instituto de Salud Carlos III MINECO and Feder Funds (FIS PI15/00974; PI18/00858 and PI18/00708) and Ministerio de Ciencia, Innovación y Universidades (PID2019-108674RB-100

Mössbauer and Magnetic Properties of Coherently Mixed Magnetite-Cobalt Ferrite Grown by Infrared Pulsed-Laser Deposition

We have studied the magnetic properties and the composition of cobalt ferrite single crystal films on SrTiO3 : Nb grown by infrared pulsed-laser deposition. Mössbauer spectra have been recorded from both the target used to grow the films and the films themselves. The Mössbauer spectra of the target taken at low temperatures show a strong dependence of the recoil free fraction of the octahedral sites with temperature. The films composition, with a coexistence of Co-enriched cobalt ferrite and magnetite, has been estimated assuming a similar ratio of the recoil free fractions of the films. X-ray absorption and x-ray magnetic circular dichroism measurements confirm the valence composition of the film and show ferromagnetic Fe-Co coupling in the films with a coercive field around 0.5 T at room temperature. The combination of these characterization techniques allows establishing the coherent structural and magnetic properties of this biphase system.(MINECO) through Projects No. MAT2012 - 38045 - C04 - 01, CTQ2013 - 43086 - P, and MAT2013 - 48009 - C4 - 1 - P and by the EU - FP7 NANOPYME Project (No. 310516).Peer Reviewe