Optimization of the Encapsulation of Vitamin D3 in Oil in Water Nanoemulsions: Preliminary Application in a Functional Meat Model System.

Meat products containing Vitamin D3 (VD3) are an innovative option that could contribute to reducing deficiencies in this micronutrient. Designing nanoemulsions that carry VD3 is the first step in developing functional meat products. Thereby, this study investigated the impact of food components on the nanoemulsion properties. A central composite design was used to study the effects of pea protein (PP, 0.5-2.5%), safflower oil (SO, 5-15%), and salt (0-0.5%) on the nanoemulsion stability (ζ-potential and particle size) and the VD3 retention. Also, the optimized nanoemulsion carrying VD3 was incorporated into a meat matrix to study its retention after cooking. The combination of food components in the optimized nanoemulsion were SO = 9.12%, PP = 1.54%, and salt content = 0.4%, resulting in the predicted values of ζ-potential, particle size, and VD3 retention of -37.76 mV, 485 nm, and 55.1%, respectively. The VD3 that was nanoencapsulated and included in a meat product remained more stable after cooking than the VD3 that was not encapsulated. If a meat product is formulated with 5 or 10% safflower oil, the stability of the nanoencapsulated VD3 is reduced. This research contributes to developing functional meat products carrying nanoencapsulated vitamin D3 in natural food-grade components.

Advances in Light-Responsive Smart Multifunctional Nanofibers: Implications for Targeted Drug Delivery and Cancer Therapy.

Over the last decade, scientists have shifted their focus to the development of smart carriers for the delivery of chemotherapeutics in order to overcome the problems associated with traditional chemotherapy, such as poor aqueous solubility and bioavailability, low selectivity and targeting specificity, off-target drug side effects, and damage to surrounding healthy tissues. Nanofiber-based drug delivery systems have recently emerged as a promising drug delivery system in cancer therapy owing to their unique structural and functional properties, including tunable interconnected porosity, a high surface-to-volume ratio associated with high entrapment efficiency and drug loading capacity, and high mass transport properties, which allow for controlled and targeted drug delivery. In addition, they are biocompatible, biodegradable, and capable of surface functionalization, allowing for target-specific delivery and drug release. One of the most common fiber production methods is electrospinning, even though the relatively two-dimensional (2D) tightly packed fiber structures and low production rates have limited its performance. Forcespinning is an alternative spinning technology that generates high-throughput, continuous polymeric nanofibers with 3D structures. Unlike electrospinning, forcespinning generates fibers by centrifugal forces rather than electrostatic forces, resulting in significantly higher fiber production. The functionalization of nanocarriers on nanofibers can result in smart nanofibers with anticancer capabilities that can be activated by external stimuli, such as light. This review addresses current trends and potential applications of light-responsive and dual-stimuli-responsive electro- and forcespun smart nanofibers in cancer therapy, with a particular emphasis on functionalizing nanofiber surfaces and developing nano-in-nanofiber emerging delivery systems for dual-controlled drug release and high-precision tumor targeting. In addition, the progress and prospective diagnostic and therapeutic applications of light-responsive and dual-stimuli-responsive smart nanofibers are discussed in the context of combination cancer therapy.

Light-Responsive and Dual-Targeting Liposomes: From Mechanisms to Targeting Strategies.

In recent years, nanocarriers have played an ever-increasing role in clinical and biomedical applications owing to their unique physicochemical properties and surface functionalities. Lately, much effort has been directed towards the development of smart, stimuli-responsive nanocarriers that are capable of releasing their cargos in response to specific stimuli. These intelligent-responsive nanocarriers can be further surface-functionalized so as to achieve active tumor targeting in a sequential manner, which can be simply modulated by the stimuli. By applying this methodological approach, these intelligent-responsive nanocarriers can be directed to different target-specific organs, tissues, or cells and exhibit on-demand controlled drug release that may enhance therapeutic effectiveness and reduce systemic toxicity. Light, an external stimulus, is one of the most promising triggers for use in nanomedicine to stimulate on-demand drug release from nanocarriers. Light-triggered drug release can be achieved through light irradiation at different wavelengths, either in the UV, visible, or even NIR region, depending on the photophysical properties of the photo-responsive molecule embedded in the nanocarrier system, the structural characteristics, and the material composition of the nanocarrier system. In this review, we highlighted the emerging functional role of light in nanocarriers, with an emphasis on light-responsive liposomes and dual-targeted stimuli-responsive liposomes. Moreover, we provided the most up-to-date photo-triggered targeting strategies and mechanisms of light-triggered drug release from liposomes and NIR-responsive nanocarriers. Lastly, we addressed the current challenges, advances, and future perspectives for the deployment of light-responsive liposomes in targeted drug delivery and therapy.

Predictive signature of response to neoadjuvant chemotherapy in muscle-invasive bladder cancer integrating mRNA expression, taxonomic subtypes, and clinicopathological features.

Background and objective: Neoadjuvant chemotherapy (NAC) followed by cystectomy is the standard of care in muscle-invasive bladder cancer (MIBC). Pathological response has been associated with longer survival, but no currently available clinicopathological variables can identify patients likely to respond, highlighting the need for predictive biomarkers. We sought to identify a predictive signature of response to NAC integrating clinical score, taxonomic subtype, and gene expression. Material and methods: From 1994 to 2014, pre-treatment tumor samples were collected from MIBC patients (stage T2-4N0/+M0) at two Spanish hospitals. A clinical score was determined based on stage, hydronephrosis and histology. Taxonomic subtypes (BASQ, luminal, and mixed) were identified by immunohistochemistry. A custom set of 41 genes involved in DNA damage repair and immune response was analyzed in 84 patients with the NanoString nCounter platform. Genes related to pathological response were identified by LASSO penalized logistic regression. NAC consisted of cisplatin/methotrexate/vinblastine until 2000, after which most patients received cisplatin/gemcitabine. The capacity of the integrated signature to predict pathological response was assessed with AUC. Overall survival (OS) and disease-specific survival (DSS) were analyzed with the Kaplan-Meier method. Results: LASSO selected eight genes to be included in the signature (RAD51, IFNγ, CHEK1, CXCL9, c-MET, KRT14, HERC2, FOXA1). The highest predictive accuracy was observed with the inclusion in the model of only three genes (RAD51, IFNɣ, CHEK1). The integrated clinical-taxonomic-gene expression signature including these three genes had a higher predictive ability (AUC=0.71) than only clinical score plus taxonomic subtype (AUC=0.58) or clinical score alone (AUC=0.56). This integrated signature was also significantly associated with OS (p=0.02) and DSS (p=0.02). Conclusions: We have identified a predictive signature for response to NAC in MIBC patients that integrates the expression of three genes with clinicopathological characteristics and taxonomic subtypes. Prospective studies to validate these results are ongoing.

Mesenchymal stem cells attenuate the proinflammatory cytokine pattern in a guinea pig model of chronic cigarette smoke exposure.

AIMS: Cigarette smoke often induces pulmonary and systemic inflammation. In animal models, mesenchymal stem cells (MSC) tend to ameliorate these effects. We aimed to explore the local and systemic expression of cytokines in guinea pigs chronically exposed to cigarette smoke, and their modifications by MSC. MAIN METHODS: Concentrations of IL-1ß, IL-6, IL-8, IL-12, TNF-α, INF-É£, TSG-6, MMP-9, TIMP-1, and/or TIMP-2 in serum and bronchoalveolar lavage (BALF) from animals exposed to tobacco smoke (20 cigarettes/day, 5 days/week for 10 weeks) were determined, and mRNA expression of some of them was measured in lung tissue. Intratracheal instillation of allogeneic bone marrow MSC (5x106 cells in 1 ml) was done at week 2. KEY FINDINGS: After cigarette smoke, IL-6 and IFN-γ increased in serum and BALF, while IL-1ß and IL-12 decreased in serum, and TSG-6 and TIMP-2 increased in BALF. IL-1ß had a paradoxical increase in BALF. MSC had an almost null effect in unexposed animals. The intratracheal administration of MSC in guinea pigs exposed to cigarette smoke was associated with a statistically significant decrease of IL-12 and TSG-6 in serum, as well as a decrease of IL-1ß and IFN-γ and an increase in TIMP-1 in BALF. Concerning mRNA expression in lung tissue, cigarette smoke did not modify the relative amount of the studied transcripts, but even so, MSC decreased the IL-12 mRNA and increased the TIMP-1 mRNA. SIGNIFICANCE: A single intratracheal instillation of MSC reduces the pulmonary and systemic proinflammatory pattern induced by chronic exposure to cigarette smoke in guinea pigs. TRIAL REGISTRATION: Not applicable.

Synthetic evolution of herbicide resistance using a T7 RNAP-based random DNA base editor.

Synthetic directed evolution via localized sequence diversification and the simultaneous application of selection pressure is a promising method for producing new, beneficial alleles that affect traits of interest in diverse species; however, this technique has rarely been applied in plants. Here, we designed, built, and tested a chimeric fusion of T7 RNA Polymerase (RNAP) and deaminase to enable the localized sequence diversification of a target sequence of interest. We tested our T7 RNAP-DNA base editor in <i>Nicotiana benthamiana</i> transient assays to target a transgene expressing <i>GFP</i> under the control of the T7 promoter and observed C-to-T conversions. We then targeted the T7 promoter-driven <i>acetolactate synthase</i> sequence that had been stably integrated in the rice genome and generated C-to-T and G-to-A transitions. We used herbicide treatment as selection pressure for the evolution of the <i>acetolactate synthase</i> sequence, resulting in the enrichment of herbicide-responsive residues. We then validated these herbicide-responsive regions in the transgenic rice plants. Thus, our system could be used for the continuous synthetic evolution of gene functions to produce variants with improved herbicide resistance.

Epithelial-to-Mesenchymal Transition Mediates Resistance to Maintenance Therapy with Vinflunine in Advanced Urothelial Cell Carcinoma.

In the phase II MAJA trial, maintenance therapy with vinflunine resulted in longer progression-free survival compared to best supportive care in advanced urothelial cell carcinoma (aUCC) patients who did not progress after first-line platinum-based chemotherapy. However, despite an initial benefit observed in some patients, unequivocal resistance appears which underlying mechanisms are presently unknown. We have performed gene expression and functional enrichment analyses to shed light on the discovery of these underlying resistance mechanisms. Differential gene expression profile of eight patients with poor outcome and nine with good outcome to vinflunine administered in the MAJA trial were analyzed. RNA was isolated from tumor tissue and gene expression was assessed by microarray. Differential expression was determined with linear models for microarray data. Gene Set Enrichment Analysis (GSEA) was used for the functional classification of the genes. In vitro functional studies were performed using UCC cell lines. Hierarchical clustering showed a differential gene expression pattern between patients with good and poor outcome to vinflunine treatment. GSEA identified epithelial-to-mesenchymal transition (EMT) as the top negatively enriched hallmark in patients with good outcome. In vitro analyses showed that the polyphenol curcumin downregulated EMT markers and sensitized UCC cells to vinflunine. We conclude that EMT mediates resistance to vinflunine and suggest that the reversion of this process could enhance the effect of vinflunine in aUCC patients.

Efecto de la reubicación de nidos en el éxito reproductivo de la tortuga marina Lepidochelys olivacea (Testudinata: Cheloniidae) / Effect of nest relocation on the reproductive success of the marine turtle Lepidochelys olivacea (Testudinata: Cheloniidae)

Resumen Introducción: Para incrementar el número de crías de tortugas marinas, es necesario mejorar estrategias de conservación, como incubación de nidos, en condiciones de vivero que contrarresten los efectos de las condiciones climáticas extremas. Objetivo: Comparar cinco parámetros de éxito reproductivo (éxito de eclosión, huevos sin desarrollo aparente, crías muertas, mortalidad embrionaria y período de incubación) de la tortuga marina Lepidochelys olivacea. Métodos: Se evaluó la incubación de nidos bajo dos períodos de reubicación de nidos (P1 ~28.8 °C agosto-septiembre y P2 ~27.1 °C octubre-noviembre, temperatura ambiental promedio, bajo condiciones de vivero, en el 2018 en Guerrero, México. Resultados: Se encontraron diferencias significativas en el éxito de eclosión y la mortalidad embrionaria entre los períodos de reubicación de nidos (P < 0.001). En P1, se observaron valores más bajos de éxito de eclosión (77.0 % P1 vs 88.6 % P2) y valores más altos de mortalidad embrionaria (13.7 % P1 vs 3.3 % P2) comparados con los de P2. Es importante señalar que, en el presente estudio la mortalidad embrionaria se presentó en una etapa tardía de desarrollo, es decir, en embriones casi completamente desarrollados. No se encontraron diferencias entre periodos respecto a los parámetros de huevos sin desarrollo aparente, crías muertas y período de incubación. Conclusiones: Las diferencias entre las condiciones ambientales durante el primer período de reubicación de nidos en comparación con el segundo período parecen afectar el éxito de eclosión y mortalidad embrionaria de L. olivacea. Por lo tanto, es importante tomar medidas al respecto durante esta etapa de incubación para tratar de mejorar la incubación de nidos bajo condiciones de vivero.

Abstract Introduction: In order to increase the number of sea turtle hatchlings, it is necessary to improve conservation strategies, such as nest incubation in hatchery conditions that counteract the effects of extreme climatic conditions. Objective: To compare five reproductive success parameters (hatching success, eggs with no apparent embryonic development, dead hatchlings, embryonic mortality, and incubation period) of the sea turtle Lepidochelys olivacea. Methods: The incubation of nests was evaluated under two periods of nest relocation, P1 ~28.8 °C (August and September) and P2 ~27.1 °C (October and November), average ambient temperature, under hatchery condition, in 2018 in Guerrero, Mexico. Results: Significant differences were found in hatching success and embryonic mortality between nest relocation periods (P < 0.001). In P1, lower values of hatching success (77.0 % P1 vs 88.6 % P2) and higher values of embryonic mortality (13.7 % P1 vs 3.3 % P2) were observed compared to those of P2. It's important pointing that, in the present study, embryonic mortality occurred at a late stage of development, that is, in almost fully developed embryos. No differences were found between periods regarding the parameters eggs with no apparent embryonic development, dead hatchlings and incubation period. Conclusions: The differences between the environmental conditions during the first period of nest relocation compared to the second period, respectively, seem to affect the hatching success and embryonic mortality of L. olivacea. Therefore, it is important to take action on this issue during this incubation stage to try to improve the incubation of nests under hatchery conditions.

Assessment of scattered radiation from hand-held dental x-ray equipment using the Monte Carlo method.

The objective of the present study was to evaluate the intensity and spatial distribution of the scattered radiation caused by the use of hand-held x-ray equipment in the zone occupied by the operator, using the Monte Carlo simulation for radiographic views of the upper and lower incisor teeth, and the consequent evaluation of the equivalent dose in the lens. In order to carry out this evaluation, the geometry of a typical dental facility with plaster walls containing the scattering object was used for the computational scenario implemented for the Monte Carlo method simulation. The PENELOPE code for Monte Carlo simulation of electron and photon transport was used with the radiation beam represented by a 60 kV spectrum, 1.5 mm Al and tungsten target. The simulations were carried out with typical parameters for workload and the number of radiographs/week. The results showed that the exposure levels varied significantly according to the angle of the x-ray beam and with the distance to the scattering object. It is concluded that the incorporation of hand-held equipment in dental radiology must be accompanied by the surveillance of occupational exposure levels and a review of the training structure of professionals in dental radiology regarding aspects of radiological protection and the particularities of using this type of equipment.

Mosquito cellular immunity at single-cell resolution.

Hemocytes limit the capacity of mosquitoes to transmit human pathogens. Here we profile the transcriptomes of 8506 hemocytes of Anopheles gambiae and Aedes aegypti mosquito vectors. Our data reveal the functional diversity of hemocytes, with different subtypes of granulocytes expressing distinct and evolutionarily conserved subsets of effector genes. A previously unidentified cell type in An. gambiae, which we term "megacyte," is defined by a specific transmembrane protein marker (TM7318) and high expression of lipopolysaccharide-induced tumor necrosis factor-α transcription factor 3 (LL3). Knockdown experiments indicate that LL3 mediates hemocyte differentiation during immune priming. We identify and validate two main hemocyte lineages and find evidence of proliferating granulocyte populations. This atlas of medically relevant invertebrate immune cells at single-cell resolution identifies cellular events that underpin mosquito immunity to malaria infection.

Vitamin D in systemic lupus erythematosus with and without lupus nephritis.

INTRODUCCIÓN: Las concentraciones séricas subóptimas de vitamina D se presentan en múltiples enfermedades crónicas, como las enfermedades autoinmunitarias. Los objetivos del estudio fueron: 1) comparar las concentraciones séricas de 25-hidroxivitamina D (25OHD3) en pacientes con lupus eritematoso sistémico (LES) con y sin nefritis lúpica (NL), y 2) evaluar la asociación de las concentraciones séricas de 25OHD3 con la actividad de la enfermedad. MATERIAL Y MÉTODOS: Estudio comparativo, transversal, que incluyó 48 mujeres con LES, con y sin NL. Se excluyeron aquellas con enfermedad renal crónica en estadio IV, cáncer, hiperparatiroidismo, embarazo o lactancia. La actividad fue evaluada con el instrumento SLEDAI-2K. La concentración sérica de 25OHD3 se determinó mediante inmunoanálisis quimioluminiscente. RESULTADOS: La media de edad de las pacientes con y sin NL fue de 36.3 ± 8.6 años y 42.7 ± 7.6 años, respectivamente. Se observó una elevada prevalencia de valores subóptimos de 25OHD3 en todas las pacientes (93%). Las concentraciones séricas de 25OHD3 fueron diferentes entre pacientes con y sin NL: 21.5 ± 6.8 ng/ml frente a 19.2 ± 7.1 ng/ml (p = 0.362). No se encontró correlación entre la concentración sérica de 25OHD3 y la actividad de la enfermedad (r = -045, p = 0.761). CONCLUSIONES: En pacientes con LES, las concentraciones séricas de 25OHD3 fueran diferentes entre pacientes con y sin NL; sin embargo, esta diferencia no fue significativa. Además, no se encontró correlación entre las concentraciones séricas de 25OHD3 y la actividad de la enfermedad evaluada por SLEDAI-2K. BACKGROUND: Sub-optimal serum vitamin D levels occur in multiple chronic diseases such as autoimmune diseases. The objectives of this study were: 1) compare the serum concentration of 25-hidroxivitamin D (25OHD3) in patients with systemic lupus erythematosus (SLE) with and without lupus nephritis (LN), and 2) evaluate the association of serum concentration of 25OHD3 with the activity of the disease. MATERIAL AND METHODS: A comparative, cross-sectional study was conducted, including 48 women with SLE, with and without clinical diagnosis of LN, according to the score of renal activity evaluated by SLEDAI-2K. Patients with stage IV chronic kidney disease, cancer, hyperparathyroidism, pregnancy and lactation were excluded. The activity was evaluated by the SLEDAI-2K instrument. The serum concentration of 25OHD3 was assessed by chemiluminescent immunoassay. RESULTS: The mean age of patients with and without LN was 36.3 ± 8.6 and 42.7 ± 7.6 years, respectively. High prevalence of suboptimal 25OHD3 levels was observed (93%). 25OHD3 concentrations were different between patients with and without LN, 21.5 ± 6.8 ng/mL vs. 19.2 ± 7.1 ng/mL (p = 0.362). No correlation was found between serum 25OHD3 concentration (r = −045, p = 0.761). CONCLUSIONS: There were no differences found in serum concentrations of 25OHD3 in patients with or without NL. Moreover, no correlation was found between serum 25OHD3 levels and the activity of the disease evaluated by SLEDAI-2K.

Performance of Flexible Chemoresistive Gas Sensors after Having Undergone Automated Bending Tests.

Many sensors are developed over flexible substrates to be used as wearables, which does not guarantee that they will actually withstand being bent. This work evaluates the gas sensing performance of metal oxide devices of three different types, before and after having undergone automated, repetitive bending tests. These tests were aimed at demonstrating that the fabricated sensors were actually flexible, which cannot be taken for granted beforehand. The active layer in these sensors consisted of WO3 nanowires (NWs) grown directly over a Kapton foil by means of the aerosol-assisted chemical vapor deposition. Their response to different H2 concentrations was measured at first. Then, they were cyclically bent, and finally, their response to H2 was measured again. Sensors based on pristine WO3-NWs over Ag electrodes and on Pd-decorated NWs over Au electrodes maintained their performance after having been bent. Ag electrodes covered with Pd-decorated NWs became fragile and lost their usefulness. To summarize, two different types of truly flexible metal oxide gas sensor were fabricated, whereas a third one was not flexible, despite being grown over a flexible substrate following the same method. Finally, we recommend that one standard bending test procedure should be established to clearly determine the flexibility of a sensor considering its intended application.

Pyrosequencing versus methylation-specific PCR for assessment of MGMT methylation in tumor and blood samples of glioblastoma patients.

Circulating biomarkers in blood may provide an interesting alternative to risky tissue biopsies in the diagnosis and follow-up of glioblastoma patients. We have assessed MGMT methylation status in blood and tissue samples from unresected glioblastoma patients who had been included in the randomized GENOM-009 trial. Paired blood and tissue samples were assessed by methylation-specific PCR (MSP) and pyrosequencing (PYR). After establishing the minimum PYR cut-off that could yield a significant difference in overall survival, we assessed the sensitivity, specificity, positive predictive value and negative predictive value (NPV) of the analyses. Methylation could be detected in cfDNA by both MSP and PYR but with low concordance with results in tissue. Sensitivity was low for both methods (31% and 38%, respectively), while specificity was higher for MSP in blood than for PYR in plasma (96% vs 76%) and NPV was similar (56 vs 57%). Concordance of results in tissue by MSP and PYR was 84.3% (P < 0.001) and correlated with outcome. We conclude that detection of cfDNA in the blood of glioblastoma patients can be an alternative when tumor tissue is not available but methods for the detection of cfDNA in blood must improve before it can replace analysis in tumor tissue.

Consenso mexicano en el manejo clínico de la hidradenitis supurativa / Mexican consensus on the clinical management of hidradenitis suppurativa

Resumen ANTECEDENTES: La hidradenitis supurativa es una enfermedad inflamatoria, crónica y recurrente con gran repercusión en la calidad de vida de los pacientes, específicamente en los aspectos psicológicos y sexuales, así como también en la productividad laboral. OBJETIVO: Generar un documento de consenso sobre el manejo clínico de los pacientes con hidradenitis supurativa en México que sirva para homologar el diagnóstico y tratamiento de estos pacientes. MÉTODO: Se convocó a un grupo multidisciplinario (dermatólogos, ginecólogos, cirujanos, infectólogos, proctólogos) para analizar la evidencia en el tratamiento integral de la hidradenitis supurativa (calidad de vida, clinimetría, diagnóstico y tratamiento), localizada mediante una revisión sistemática de la bibliografía. El grupo discutió sobre los temas y se generaron las recomendaciones por acuerdo unánime de los participantes. RESULTADOS: Con frecuencia los pacientes con hidradenitis supurativa pueden tardar mucho tiempo en obtener el diagnóstico y manejo correctos, principalmente debido a la falta de evidencia sobre la enfermedad, así como a la falta de criterios para referir al paciente con un especialista. Se emitieron 22 recomendaciones para el tratamiento de pacientes con hidradenitis supurativa, que comprenden el diagnóstico, estadificación y clinimetría, así como el manejo clínico en el primer, segundo y tercer nivel de atención. CONCLUSIONES: Se emitieron las recomendaciones con base en la mejor evidencia disponible, así como la experiencia del grupo multidisciplinario de expertos en el tratamiento de hidradenitis supurativa.

Abstract BACKGROUND: Hidradenitis suppurativa is an inflammatory, chronic and recurrent disease with a high impact on patients quality of life, specifically in psychological and sexual aspects, as well as in labor productivity. OBJECTIVE: To generate a consensus document on the clinical management of patients with hidradenitis suppurativa in Mexico, that serves to standardize the diagnosis and treatment of these patients. METHOD: A multidisciplinary group was convened (dermatologists, gynecologists, surgeons, infectologists, proctologists) to analyze the evidence on the integral treatment of hidradenitis suppurativa (quality of life, clinimetry, diagnosis and treatment), located through a systematic review of the literature. The group discussed the issues and the recommendations were generated by unanimous agreement of the participants. RESULTS: Frequently, patients with hidradenitis suppurativa can take a long time to obtain the correct diagnosis and management, mainly due to the lack of evidence about the disease, as well as the lack of criteria to refer the patient to a specialist. Twenty-two recommendations were issued for the treatment of patients with hidradenitis suppurativa, which include diagnosis, staging and clinimetry, as well as clinical management in the first, second and third levels of care. CONCLUSIONS: The recommendations were issued based on the best available evidence, as well as the experience of the multidisciplinary group of experts in the treatment of hidradenitis suppurativa.

Correction: Monitoring EGFR-T790M mutation in serum/plasma for prediction of response to third-generation EGFR inhibitors in patients with lung cancer.

[This corrects the article DOI: 10.18632/oncotarget.25478.].

Association of PALB2 Messenger RNA Expression with Platinum-Docetaxel Efficacy in Advanced Non-Small Cell Lung Cancer.

INTRODUCTION: Partner and localizer of BRCA2 (PALB2) is essential for homologous recombination repair. We examined mRNA levels of DNA repair genes, including partner and localizer of BRCA2 gene (PALB2), ring finger protein 8 gene (RNF8), replication timing regulatory factor 1 gene (RIF1), ATM serine/threonine kinase gene (ATM), and tumor protein p53 binding protein 1 gene (53BP1) as predictive biomarkers for cisplatin-docetaxel in the European phase III BRCA1, DNA repair associated (BRCA1)-receptor-associated protein 80 (RAP80) expression customization (BREC) phase III clinical trial (ClinicalTrials.gov identifier NCT00617656). METHODS: The study was a prespecified secondary objective of the BREC trial. We assessed mRNA levels of PALB2 and four more DNA repair genes (RNF8, RIF1, ATM and 53BP1) as biomarkers in tissue from 177 patients with cisplatin-docetaxel-treated NSCLC. We examined the relationship of gene expression levels with progression-free survival, overall survival, and response. RESULTS: In 177 patients with NSCLC (who had a median age of 62 years and included 140 men and 91 patients with adenocarcinoma), only high PALB2 mRNA expression was predictive in the progression-free survival Cox regression analysis (hazard ratio = 0.63, 95% confidence interval: 0.42-0.83, p = 0.0080). PALB2 was also predictive of overall survival (hazard ratio = 0.68, 95% confidence interval: 0.42-0.90, p = 0.0266). Among the 158 patients evaluable for response, high PALB2 mRNA expression was predictive of response to cisplatin-docetaxel. Specifically, an objective response rate of 77% to cisplatin-docetaxel was observed for patients with high PALB2 mRNA expression compared with a rate of only 23 % for those with low PALB2 mRNA expression (p = 0.0448). CONCLUSIONS: High PALB2 mRNA expression identified patients with NSCLC who significantly benefited from cisplatin-docetaxel chemotherapy in the European BREC phase III clinical trial. The combination of chemotherapy with immunotherapy will become the standard of care, and a predictive marker of response to chemotherapy may accurately guide therapeutic decision making.

5 protein-based signature for resectable lung squamous cell carcinoma improves the prognostic performance of the TNM staging.

INTRODUCTION: Prognostic biomarkers have been very elusive in the lung squamous cell carcinoma (SCC) and none is currently being used in the clinical setting. We aimed to identify and validate the clinical utility of a protein-based prognostic signature to stratify patients with early lung SCC according to their risk of recurrence or death. METHODS: Patients were staged following the new International Association for the Study of Lung Cancer (IASLC) staging criteria (eighth edition, 2018). Three independent retrospective cohorts of 117, 96 and 105 patients with lung SCC were analysed to develop and validate a prognostic signature based on immunohistochemistry for five proteins. RESULTS: We identified a five protein-based signature whose prognostic index (PI) was an independent and significant predictor of disease-free survival (DFS) (p<0.001; HR=4.06, 95% CI 2.18 to 7.56) and overall survival (OS) (p=0.004; HR=2.38, 95% CI 1.32 to 4.31). The prognostic capability of PI was confirmed in an external multi-institutional cohort for DFS (p=0.042; HR=2.01, 95% CI 1.03 to 3.94) and for OS (p=0.031; HR=2.29, 95% CI 1.08 to 4.86). Moreover, PI added complementary information to the newly established IASLC TNM 8th edition staging system. A combined prognostic model including both molecular and anatomical (TNM) criteria improved the risk stratification in both cohorts (p<0.05). CONCLUSION: We have identified and validated a clinically feasible protein-based prognostic model that complements the updated TNM system allowing more accurate risk stratification. This signature may be used as an advantageous tool to improve the clinical management of the patients, allowing the reduction of lung SCC mortality through a more accurate knowledge of the patient's potential outcome.

Genomic profiling in advanced stage non-small-cell lung cancer patients with platinum-based chemotherapy identifies germline variants with prognostic value in SMYD2.

OBJECTIVE: The aim of the study was to investigate the relationship between germline variations as a prognosis biomarker in patients with advanced Non-Small-Cell-Lung-Cancer (NSCLC) subjected to first-line platinum-based treatment. MATERIALS AND METHODS: We carried out a two-stage genome-wide-association study in non-small-cell lung cancer patients with platinum-based chemotherapy in an exploratory sample of 181 NSCLC patients from Caucasian origin, followed by a validation on 356 NSCLC patients from the same ancestry (Valencia, Spain). RESULTS: We identified germline variants in SMYD2 as a prognostic factor for survival in patients with advanced NSCLC receiving chemotherapy. SMYD2 alleles are associated to a decreased overall survival and with a reduced Time to Progression. In addition, enrichment pathway analysis identified 361 variants in 40 genes to be involved in poorer outcome in advanced-stage NSCLC patients. CONCLUSION: Germline SMYD2 alleles are associated with bad clinical outcome of first-line platinum-based treatment in advanced NSCLC patients. This result supports the role of SMYD2 in the carcinogenic process, and might be used as prognostic signature directing patient stratification and the choice of therapy. MICROABSTRACT: A two-Stage Genome wide association study in Caucasian population reveals germline genetic variation in SMYD2 associated to progression disease in first-line platinum-based treatment in advanced NSCLC patients. SMYD2 profiling might have prognostic / predictive value directing choice of therapy and enlighten current knowledge on pathways involved in human carcinogenesis as well in resistance to chemotherapy.

Monitoring EGFR-T790M mutation in serum/plasma for prediction of response to third-generation EGFR inhibitors in patients with lung cancer.

BACKGROUND: Osimertinib is efficacious in lung cancer patients with epidermal growth factor receptor (EGFR) mutations and acquired resistance (AR) to EGFR tyrosine kinase inhibitors due to EGFR-T790M mutation (T790M). We sought to describe T790M changes in serum/plasma during osimertinib therapy and correlate these changes with treatment outcomes. MATERIAL AND METHODS: Serum/plasma from EGFR-mutant lung cancer patients with T790M-AR was collected before and during osimertinib treatment. Changes in T790M were evaluated using a peptide-nucleic acid-PCR assay, and correlated with clinical and radiographic response. RESULTS: Thirteen patients were included. Median time on osimertinib treatment was 10.6 months with a median progression-free survival of 13.6 months. Best response to osimertinib was partial response (PR), stable disease (SD) or progression (PD) in 46.1%, 30.8% and 23.1% of patients, respectively.Most of the patients were paucisymptomatic at baseline. Symptom improvement was reported in 66.6% of responder patients; while symptoms remained stable in 75% of patients with SD, and 66% of patients with PD had clinical deterioration.Three patterns of T790M changes during osimertinib treatment were identified. T790 remained detectable with PD or a short-lasting SD in 15.4% of the patients. T790M disappeared in 69.2% of patients with PR or SD. T790M disappeared, despite clinical and/or radiographic progression in 15.4% of the patients. CONCLUSION: Changes of T790M in serum/plasma in EGFR-mutant lung cancer patients with T790M-AR might be a useful marker of symptomatic and radiographic outcome to osimertinib. Longer follow-up is needed to establish if subsequent emergence of T790M could be a marker of resistance.