Impact of Wnt/β-Catenin Inhibition on Cell Proliferation through CDC25A Downregulation in Soft Tissue Sarcomas.

The Wnt signaling pathway is an important cellular mechanism for regulating differentiation processes as well as cell cycle events, and different inhibitors of this pathway, for example, PRI-724, are showing promising results in clinical trials for treatment of advanced pancreatic adenocarcinoma or ovarian cancer. Growing evidence suggests that Wnt signaling may also be crucial for tumorigenesis and progression of soft tissue sarcomas (STS), a malignant neoplasm with few therapeutic options at an advanced state. Our study with several STS cell lines and primary cultures shows that inhibition of Wnt/β-catenin signaling with PRI-724 is able to suppress cell viability/proliferation and to increase cell death rates. TCF/β-catenin-mediated transcriptional activity is decreased in treated cells, leading to downregulation of its target genes CCND1 and CDC25A. The latter was critical because its downregulation via siRNA was able to mimic the effect of PRI-724 on cell cycle arrest and cell death induction. An evaluation of NCBI/GenBank data confirmed that CDC25A mRNA is elevated in STS patients. Importantly, PRI-724 in combination with standard STS chemotherapeutics doxorubicin or trabectedin enhanced their antitumoral effect in a synergistic manner according to isobolographic analysis, suggesting that Wnt inhibition through PRI-724 could be a beneficial combination regime in patients with advanced STS.This study was financed by Grupo Español de Investigación en Sarcomas (GEIS) and Fundación Mari PazJiménez Casado. MPC is supported by Programa Estrategia de Emprendimiento y Empleo Joven,Garantía Juvenil(Ministerio de Trabajo, Migraciones y Seguridad Social-SOIB.S

Intratumoral immunosuppression profiles in 11q-deleted neuroblastomas provide new potential therapeutic targets

High‐risk neuroblastoma (NB) patients with 11q deletion frequently undergo late but consecutive relapse cycles with fatal outcome. To date, no actionable targets to improve current multi‐modal treatment have been identified. We analyzed immune microenvironment and genetic profiles of high‐risk NB correlating with 11q immune status. We show in two independent cohorts that 11q‐deleted NB exhibit various immune‐inhibitory mechanisms, including increased CD4+ resting T cells and M2 macrophages, higher expression of programmed death‐ligand 1, interleukin‐10, transforming growth factor‐beta‐1 and indoleamine 2,3‐dioxygenase 1 (P<0.05), and also higher chromosomal breakages (P≤0.02) and hemizygosity of immunosuppressive miRNAs than MYCN‐amplified and other 11q‐non‐deleted high‐risk NB. We also analyzed benefits of maintenance treatment in 83 high‐risk stage M NB patients focusing on 11q status, either with standard anti‐GD2 immunotherapy (n=50) or previous retinoic acid‐based therapy alone (n=33). Immunotherapy associated with higher EFS (50 vs. 30, P=0.028) and OS (72 vs. 52, P=0.047) at 3 years in the overall population. Despite benefits from standard anti‐GD2 immunotherapy in high‐risk NB patients, those with 11q deletion still face poor outcome. This NB subgroup displays intratumoral immune suppression profiles, revealing a potential therapeutic strategy with combination immunotherapy to circumvent this immune checkpoint blockade

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

Spatiotemporal Characteristics of the Largest HIV-1 CRF02_AG Outbreak in Spain: Evidence for Onward Transmissions

Background and Aim: The circulating recombinant form 02_AG (CRF02_AG) is the predominant clade among the human immunodeficiency virus type-1 (HIV-1) non-Bs with a prevalence of 5.97% (95% Confidence Interval-CI: 5.41–6.57%) across Spain. Our aim was to estimate the levels of regional clustering for CRF02_AG and the spatiotemporal characteristics of the largest CRF02_AG subepidemic in Spain.Methods: We studied 396 CRF02_AG sequences obtained from HIV-1 diagnosed patients during 2000–2014 from 10 autonomous communities of Spain. Phylogenetic analysis was performed on the 391 CRF02_AG sequences along with all globally sampled CRF02_AG sequences (N = 3,302) as references. Phylodynamic and phylogeographic analysis was performed to the largest CRF02_AG monophyletic cluster by a Bayesian method in BEAST v1.8.0 and by reconstructing ancestral states using the criterion of parsimony in Mesquite v3.4, respectively.Results: The HIV-1 CRF02_AG prevalence differed across Spanish autonomous communities we sampled from (p &lt; 0.001). Phylogenetic analysis revealed that 52.7% of the CRF02_AG sequences formed 56 monophyletic clusters, with a range of 2–79 sequences. The CRF02_AG regional dispersal differed across Spain (p = 0.003), as suggested by monophyletic clustering. For the largest monophyletic cluster (subepidemic) (N = 79), 49.4% of the clustered sequences originated from Madrid, while most sequences (51.9%) had been obtained from men having sex with men (MSM). Molecular clock analysis suggested that the origin (tMRCA) of the CRF02_AG subepidemic was in 2002 (median estimate; 95% Highest Posterior Density-HPD interval: 1999–2004). Additionally, we found significant clustering within the CRF02_AG subepidemic according to the ethnic origin.Conclusion: CRF02_AG has been introduced as a result of multiple introductions in Spain, following regional dispersal in several cases. We showed that CRF02_AG transmissions were mostly due to regional dispersal in Spain. The hot-spot for the largest CRF02_AG regional subepidemic in Spain was in Madrid associated with MSM transmission risk group. The existence of subepidemics suggest that several spillovers occurred from Madrid to other areas. CRF02_AG sequences from Hispanics were clustered in a separate subclade suggesting no linkage between the local and Hispanic subepidemics

Estudi de la via de WNT com a diana terapèutica en tumors sòlids i identificació de nous marcadors pronòstics i predictius

[cat] La via de senyalització Wnt està implicada en la regulació de processos importants en la progressió del càncer com: el creixement tumoral, la senescència, la diferenciació i la metàstasi. El seu estudi ha contribuït tant per a millorar la comprensió de la carcinogènesi, sent el càncer colorectal el millor paradigma de l’estudi de la progressió tumoral iniciada en gran part per alteracions en aquesta via de senyalització, com pel desenvolupament d’inhibidors de la via per tal de millorar les estratègies terapèutiques actuals. En aquesta tesi es va estudiar la implicació de la via Wnt/β- catenina en dos tipus de tumors sòlids diferents, els sarcomes de parts toves i el càncer de pròstata avançat. Els sarcomes de parts toves (SPT) són un grup de tumors poc freqüents i molt heterogeni. L’etiologia d’aquests tumors segueix sent desconeguda i malgrat la seva heterogeneïtat se’ls considera com una sola entitat respecte a diagnòstic i estratègies terapèutiques. En aquest sentit, estudis enfocats a comprendre millor el procés pel qual s’originen aquests tipus de neoplàsies i la identificació de possibles dianes terapèutiques que aportin millores en el maneig d’aquests pacients són essencials. Dins aquest context, en aquesta tesi es va estudiar la implicació de la via Wnt/β- catenina en la sarcomagènesi i la seva inhibició com a possible estratègia terapèutica. S’ha demostrat que la via de senyalització Wnt està activada en línies cel·lulars i cultius primaris representatius de sarcomes de parts toves, promovent l’expressió de gens diana de la via, importants per la proliferació cel·lular, en concret del gen CDC25A. La inhibició de la via, quan s’actua per sota de β-catenina (proteïna clau de la via) en la cascada de senyalització, redueix la proliferació cel·lular provocant que les cèl·lules entrin en apoptosi, a través de la inhibició dels nivells d’expressió de CDC25A. De manera interessant, la combinació d’aquests inhibidors amb els agents quimioterapèutics convencionals millora el seu efecte antitumoral d’una manera sinèrgica, obrint les portes a la realització d’estudis en models experimentals in vivo. Per altra banda, l’estudi de biomarcadors predictius i/o pronòstics de resposta o progressió a determinats tractaments, s’ha convertit en un tema recurrent en la investigació contra el càncer. En primer lloc, perquè permeten la identificació de noves dianes terapèutiques per a les quals desenvolupar fàrmacs que millorin els tractaments actuals. En aquest sentit, els resultats obtinguts en aquesta tesi en mostres tumorals de pacients amb SPT, estableixen les bases per identificar alteracions moleculars de la via Wnt/β-catenina per a ser usades com a biomarcadors de resposta a determinats inhibidors en assaigs clínics, com és el cas del paper clau de CDC25A en la proliferació cel·lular. I en segon lloc, perquè durant el tractament apareixen mecanismes de resistència que provoquen la progressió cap una forma de la malaltia més avançada, com en el cas del càncer de pròstata avançat. En aquests tipus de tumors s’ha descrit que la via Wnt/β-catenina hi juga un paper important, però encara es desconeix, en gran mesura, el mecanisme amb què ho fa. En relació amb això en aquest treball, s’han identificat determinats miRNAs (hsa-miR-205-5p i hsa-miR-1301- 3p) que podrien estar regulant a components de la via Wnt/β-catenina durant la progressió de la malaltia. Aquests resultats constitueixen la base per avaluar en futurs estudis si la determinació d’aquests nous biomarcadors, en biòpsia líquida i de forma prospectiva, podrien complementar o millorar el test del PSA (de l’anglès, prostate specific antigen) en sang, el biomarcador utilitzat actualment en càncer de pròstata.[spa] La vía de señalización Wnt está implicada en la regulación de procesos importantes para la progresión del cáncer como: el crecimiento tumoral, la senescencia, la diferenciación y la metástasis. Su estudio ha contribuido a mejorar la comprensión de la carcinogénesis, siendo el cáncer de colon, el mejor paradigma tanto para el estudio de la progresión tumoral iniciada en gran parte por alteraciones en esta vía de señalización, así como al desarrollo de inhibidores de la vía con el objetivo de mejorar las estrategias terapéuticas actuales. En la presente tesis se estudió la implicación de la vía Wnt/β-catenina en dos tipos de tumores diferentes, los sarcomas de partes blandas y el cáncer de próstata avanzado. Los sarcomas de partes blandas (SPB) son un grupo muy heterogéneo, de tumores poco frecuentes. La etiología de estos tumores sigue siendo desconocida y a pesar de su heterogeneidad se los considera como una sola entidad en cuanto al diagnóstico y estrategias terapéuticas. En este sentido, estudios enfocados en mejorar la comprensión del proceso por el cual se originan este tipo de neoplasias y la identificación de posibles dianas terapéuticas que aporten mejoras en el manejo de estos pacientes son esenciales. Dentro de este contexto, en esta tesis se estudió la implicación de la vía Wnt/β-catenina en la sarcomagénesis y su inhibición como posible estrategia terapéutica. Se ha demostrado que la vía de señalización Wnt está activada en líneas celulares y cultivos primarios representativos de sarcomas de partes blandas, promoviendo la expresión de genes diana de la vía importantes para la proliferación celular, en concreto, de CDC25A. La inhibición de la vía, actuando per debajo de β- catenina (proteína clave de la vía) en la cascada de señalización, reduce la proliferación celular haciendo que las células entren en apoptosis, a través de la inhibición de los niveles de expresión de CDC25A. De manera interesante, la combinación de estos inhibidores con los quimioterapéuticos convencionales mejora su efecto antitumoral, de forma sinérgica, abriendo las puertas a la realización de nuevos experimentos en modelos in vivo. Por otro lado, el estudio de biomarcadores predictivos y/o pronósticos de respuesta o progresión a determinados tratamientos, se ha convertido en un tema recurrente en la investigación contra el cáncer. En primer lugar, porque permiten la identificación de nuevas dianas terapéuticas, para las cuales desarrollar nuevos fármacos que mejoren los tratamientos actuales. En este sentido, los resultados obtenidos en esta tesis en muestras tumorales de pacientes con SPB, sirven de base para identificar alteraciones moleculares de la vía Wnt/β-catenina que podrían ser usadas como biomarcadores de respuesta a determinados inhibidores en ensayos clínicos, como en el caso del papel clave de CDC25A en la proliferación celular. Y en segundo lugar, porque durante el tratamiento aparecen mecanismos de resistencia que provocan la progresión a una forma de la enfermedad más avanzada, como en el caso del cáncer de próstata avanzado. En este tipo de tumores se ha descrito que la vía Wnt/β-catenina juega un papel importante, aunque todavía se desconoce, en gran medida, la manera en que lo hace. En línea con esto en este trabajo, se han identificado determinados miRNAs (hsamiR- 205-5p y hsa-miR-1301-3p) que podrían estar regulando a componentes de la vía Wnt/β-catenina durante la progresión de la enfermedad. Estos resultados sirven de base para evaluar en futuros estudios si la detección de estos nuevos biomarcadores, en biopsia líquida de forma prospectiva, pudiera complementar o mejorar el test de PSA en sangre, el biomarcador usado actualmente en cáncer de próstata.[eng] Wnt signalling pathway regulates various processes that are important for cancer progression, including: tumor initiation, tumor growth, cell senescence, cell death, differentiation and metastasis. The study of this pathway has improved the understanding of carcinogenesis, especially in colon cancer, the paradigm of studies about tumor progression initiated through alterations in Wnt signalling pathway. Moreover, it has contributed to improving current therapeutic strategies by the development of small inhibitors targeting this pathway. In this thesis, we evaluate the role of Wnt/β-catenin pathway in two different types of tumours: soft tissue sarcoma and advanced prostate cancer. Soft tissue sarcomas (STS) are a group of rare and heterogeneous tumours. The aetiology of these tumours is still unknown and despite their heterogeneity they are considered as a single entity in their diagnosis and therapeutic approach. In this context, research focusing on the study of new molecular pathways that provide a better understanding of sarcomagenesis and identification of potential therapeutic targets to improve the management of these patients are a primordial necessity. In this thesis, we evaluated the role of Wnt/β-catenin pathway in sarcomagenesis and its inhibition as a potential therapeutic strategy. We demonstrated that Wnt signalling pathway is activated in a representative panel of STS cell lines and tumour-derived cells, promoting the expression of target genes mediators of cell proliferation, such as CDC25A. Inhibition of the pathway, β-catenin-downstream, reduced cell proliferation by inducing apoptosis, through downregulation of CDC25A expression. Interestingly, the combination of these inhibitors with conventional chemotherapeutic agents enhances their antitumor effect in a synergistic manner, opening the door for further experiments using in vivo models. On the other hand, identification of predictive/prognostic biomarkers of response or treatment progression has become a recurring topic in cancer research. First, because it serves as the basis of studies related to the identification of new therapeutic targets for the improvement of current treatments. In line with this, the results presented in this thesis, by analyzing tumor samples of patients with STS, serve as a basis for further studies to evaluate the role of molecular alterations of the Wnt/β-catenin pathway as predictive biomarkers of response to inhibitors which are in clinical trials, as in the case of the key role of CDC25A in cell proliferation. And secondly, due to the treatment resistance that all patients eventually develop, as in advanced prostate cancer. It has been suggested that Wnt/β-catenin pathway plays an important role, during androgen deprivation therapy progression, although the mechanics behind it is still unknown. In line with this we identified certain miRNAs (hsa-miR-205-5p and hsa-miR-1301-3p) that could be implicated in the disease progression by regulating components of the Wnt/β-catenin pathway. Further studies will evaluate whether the detection of these new biomarkers, in liquid biopsy, could complement or improve the currently used biomarker in prostate cancer (PSA)

WNT/β-Catenin Pathway in Soft Tissue Sarcomas: New Therapeutic Opportunities?

Soft tissue sarcomas (STS) are a very heterogeneous group of rare tumors, comprising more than 50 different histological subtypes that originate from mesenchymal tissue. Despite their heterogeneity, chemotherapy based on doxorubicin (DXR) has been in use for forty years now and remains the standard first-line treatment for locally advanced unresectable or metastatic STS, although overall survival could not be improved by combination with other chemotherapeutics. In this sense, the development of new therapeutic approaches continues to be a largely unmatched goal. The WNT/β-catenin signaling pathway is involved in various fundamental processes for embryogenic development, including the proliferation and differentiation of mesenchymal stem cells. Although the role of this pathway has been widely researched in neoplasms of epithelial origin, little is known about its relevance for mesenchymal neoplasms. This review covers the most important molecular alterations of the WNT signaling pathway in STS. The detection of these alterations and the understanding of their functional consequences for those pathways controlling sarcomagenesis development and progression are crucial to broaden the current knowledge about STS as well as to identify novel drug targets. In this regard, the current therapeutic options and drug candidates to modulate WNT signaling, which are usually classified by their interaction site upstream or downstream of β-catenin, and their presumable clinical impact on STS are also discussed

TLR/WNT: A Novel Relationship in Immunomodulation of Lung Cancer

The most frequent cause of death by cancer worldwide is lung cancer, and the 5-year survival rate is still very poor for patients with advanced stage. Understanding the crosstalk between the signaling pathways that are involved in disease, especially in metastasis, is crucial to developing new targeted therapies. Toll-like receptors (TLRs) are master regulators of the immune responses, and their dysregulation in lung cancer is linked to immune escape and promotes tumor malignancy by facilitating angiogenesis and proliferation. On the other hand, over-activation of the WNT signaling pathway has been reported in lung cancer and is also associated with tumor metastasis via induction of Epithelial-to-mesenchymal-transition (EMT)-like processes. An interaction between both TLRs and the WNT pathway was discovered recently as it was found that the TLR pathway can be activated by WNT ligands in the tumor microenvironment; however, the implications of such interactions in the context of lung cancer have not been discussed yet. Here, we offer an overview of the interaction of TLR-WNT in the lung and its potential implications and role in the oncogenic process

Disruption of TCF/β-Catenin Binding Impairs Wnt Signaling and Induces Apoptosis in Soft Tissue Sarcoma Cells.

Soft tissue sarcomas (STS) are malignant tumors of mesenchymal origin and represent around 1% of adult cancers, being a very heterogeneous group of tumors with more than 50 different subtypes. The Wnt signaling pathway is involved in the development and in the regulation, self-renewal, and differentiation of mesenchymal stem cells, and plays a role in sarcomagenesis. In this study, we have tested pharmacologic inhibition of Wnt signaling mediated by disruption of TCF/β-catenin binding and AXIN stabilization, being the first strategy more efficient in reducing cell viability and downstream effects. We have shown that disruption of TCF/β-catenin binding with PKF118-310 produces in vitro antitumor activity in a panel of prevalent representative STS cell lines and primary cultures. At the molecular level, PKF118-310 treatment reduced β-catenin nuclear localization, reporter activity, and target genes, resulting in an increase in apoptosis. Importantly, combination of PKF118-310 with doxorubicin resulted in enhanced reduction of cell viability, suggesting that Wnt inhibition could be a new combination regime in these patients. Our findings support the usefulness of Wnt inhibitors as new therapeutic strategies for the prevalent STS. Mol Cancer Ther; 16(6); 1166-76. ©2017 AACR

RG7112, a Small-Molecule Inhibitor of MDM2, Enhances Trabectedin Response in Soft Tissue Sarcomas

<div><p>ABSTRACT</p><p>MDM2 is a critical negative regulator of the p53 tumor suppressor protein. Selected sarcoma subtypes are being treated with Trabectedin in second line, which promotes DNA damage and p53-dependent apoptosis. The aim of this study was to evaluate the improvement of Trabectedin response with MDM2 inhibitors in soft tissue sarcomas. The antitumor effects of Trabectedin, Nutlin-3A and RG7112 as single agents or in combination were examined <i>in vitro</i>. RG7112 significantly synergized with Trabectedin in MDM2-amplified liposarcoma cells, representing a promising new therapeutic strategy for the treatment of sarcomas with MDM2 amplification.</p></div