Differences in expression profiling and biomarkers between histological colorectal carcinomas subsets from the serrated pathway.

Histological subtypes of colorectal carcinomas (CRCs) arising from the serrated route, such as serrated adenocarcinoma (SAC) and CRC showing histological and molecular features of microsatellite instability (hmMSI-H), share common features (female gender, right-sided location, mucinous histology and altered CpG methylation) but dramatically differ in terms of prognosis, development of immune response and treatment options. Despite this, to date no expression profiling comparison has been carried out for finding out functions and molecules responsible for such differences. METHODS AND RESULTS: Molecular signatures of SAC and hmMSI-H were obtained by transcriptomic array; qPCR and immunohistochemistry (IHC) were used to validate differentially expressed genes. An over-representation of innate immunity functions (granulomonocytic recruitment, chemokine production, TLR signaling, antigen processing and presentation) were obtained from this comparison and ICAM1 was more expressed in hmMSI-H whereas two genes (CRCP and CXCL14) were more expressed in SAC. These array results were subsequently validated by qPCR, and CXCL14 and ICAM1 by IHC. Information retrieved from public databanks confirmed our findings. CONCLUSIONS: Our findings highlight specific functions and genes which provide a better understanding of the role of the immune response in the serrated pathological route and may be of help in identifying actionable molecules. This article is protected by copyright. All rights reserved.pre-print664 K

Effects of imipramine on cancer patients over-expressing Fascin1; description of the HITCLIF clinical trial

BackgroundTumor invasion and metastasis are responsible for the majority of cancer-related deaths. The identification of molecules involved in these processes is crucial to design effective treatments that can halt the progression of cancer. To spread and metastasize, tumor cells must restructure their cytoskeleton and emit protrusions. A key molecule in this process of creating these invading structures is Fascin1, the main protein involved in the formation of actin cytoskeleton bundles and a consistent marker of bad prognosis in several types of cancer. Recent studies have shown that imipramine, an FDA- and EMA-approved antidepressant, can block Fascin1and prevent the formation of actin bundles, making it a promising candidate for the treatment of Fascin1-expressing cancers. As a result, a clinical trial will be conducted to assess the efficacy of imipramine being the first experimental clinical study selecting patients based on Fascin1 expression.MethodsThe HITCLIF trial is a multicenter, double-blind, placebo-controlled, randomized and non-commercial phase II clinical trial conducted in parallel groups to evaluate the effectiveness of the tricyclic antidepressant imipramine as anti-invasive agent in the treatment of localized colon, rectal and triple negative breast cancer patients with overexpression of Fascin1. Eligible patients will be randomly assigned, in a 1:1 ratio, to receive imipramine or placebo. Patients will be stratified into 2 groups according to whether administration of imipramine is concomitant with neoadjuvant chemotherapy regimen. Group A will receive imipramine alone without neoadjuvant chemotherapy, while Group B will receive imipramine treatment along with the standard neoadjuvant chemotherapy regimen. The primary endpoint of the trial is the grade of alteration in the prognostic histopathological features at invasive margins (tumor budding, cytoplasmic pseudo-fragments, tumor growth pattern, and peritumoral lymphocytic infiltration).DiscussionFascin1 is an interesting therapeutical target as it plays a causative role in the invasion and metastasis of cancer cells. Moreover, its expression is virtually absent in normal epithelia but highly expressed in cancer with bad prognosis. In silico, in vitro and in vivo studies by our group have demonstrated that the antidepressant imipramine has Fascin1-dependant anti-invasive and anti-metastatic effects in colorectal cancer cells. Now we are recruiting patients in a clinical trial based on Fascin1 over-expression in which administration of imipramine will be carried out during the period between the diagnosis biopsy and surgical resection to explore the drug effects on tumor invasive front.Clinical trial registrationhttps:///www.clinicaltrialsregister.eu/ctr-search/trial/2021-001328-17/ES, identifier 2021-001328-17

Methylome profiling reveals functions and genes which are differentially methylated in serrated compared to conventional colorectal carcinoma.

Background: Serrated adenocarcinoma (SAC) is a recently recognized colorectal cancer (CRC) subtype accounting for 7.5–8.7 % of CRCs. It has been shown that SAC has a worse prognosis and different histological and molecular features compared to conventional carcinoma (CC) but, to date, there is no study analysing its methylome profile. Results: The methylation status of 450,000 CpG sites using the Infinium Human Methylation 450 BeadChip array was investigated in 103 colorectal specimens, including 39 SACs and 34 matched CCs, from Spanish and Finnish patients. Microarray data showed a higher representation of morphogenesis-, neurogenesis-, cytoskeleton- and vesicle transport-related functions and also significant differential methylation of 15 genes, including the iodothyronine deiodinase DIO3 and the forkhead family transcription factor FOXD2 genes which were validated at the CpG, mRNA and protein level using pyrosequencing, methylation-specific PCR, quantitative polymerase chain reaction (qPCR) and immunohistochemistry. A quantification study of the methylation status of CpG sequences in FOXD2 demonstrated a novel region controlling gene expression. Moreover, differences in these markers were also evident when comparing SAC with CRC showing molecular and histological features of high-level microsatellite instability. Conclusions: This methylome study demonstrates distinct epigenetic regulation patterns in SAC which are consistent to previous expression profile studies and that DIO3 and FOXD2 might be molecular targets for a specific histologyoriented treatment of CRC.post-print2306 K

Study of IDO1 gene expression in histological variants of colorectal carcinoma.

pre-print477 K

Método para la detección y determinación de carga viral del virus del papiloma humano

Número de publicación: 2 357 821 Número de solicitud: 200930425Un par de cebadores degenerados capaces de amplificar un fragmento del gen L1 de varios genotipos del virus del papiloma humano (VPH) y un método de detección y de cuantificación de la carga viral del VPH en muestras biológicas aisladas, que comprende: la extracción de ADN de una muestra biológica aislada, poner en contacto la muestra con una mezcla de reacción que contiene al menos un fluorocromo y los cebadores degenerados de la invención, amplificar un fragmento del gen L1 del VPH, medir la fluorescencia obtenida en el paso de amplificación y comparar el valor de fluorescencia obtenido con una secuencia control.Universidad Politécnica de CartagenaFundación para la Formación e Investigación Sanitarias de la Región de Murci

The effect of fascin 1 inhibition on head and neck squamous cell carcinoma cells

Fascin 1 plays important pro-metastatic roles in head and neck carcinoma (HNSCC) migration, invasion, and metastasis. However, limited advancement in targeting metastasis remains a major obstacle in improving HNSCC patients' survival. Therefore, we assessed the therapeutic potential of fascin 1 targeted inhibition and its potential prognostic value in HNSCC patients. Using in vitro and in vivo approaches, we investigated the effect of compound G2, a novel fascin 1 inhibitor, on HNSCC cells migration, invasion, and metastasis. High-throughput screening (HTS) was used to assess cytotoxic activity of compound G2 alone or combined with irradiation. We also evaluated the prognostic potential of fascin 1 in HNSCC patients. Interestingly, compound G2 reduced carcinoma cells migration and invasion in vitro and inhibited metastasis in vivo. Moreover, HTS revealed a modest cytotoxic activity of the compound G2 on HNSCC cell lines. Irradiation did not synergistically enhance the compound G2-mediated cytotoxic activity. Survival analyses showed that high fascin 1 immunoexpression, at the tumor invasive front, was associated with cancer-specific mortality in the advanced stages of HNSCC. Collectively, our findings suggest that fascin 1 represents a promising anti-metastatic therapeutic target and a useful prognostic marker in patients with HNSCC. Novel anti-metastatic agents could provide a valuable addition to cancer therapy.Peer reviewe

Novel anti-invasive properties of a Fascin1 inhibitor on colorectal cancer cells

Tumor invasion and metastasis involve processes in which actin cytoskeleton rearrangement induced by Fascin1 plays a crucial role. Indeed, Fascin1 has been found overexpressed in tumors with worse prognosis. Migrastatin and its analogues target Fascin1 and inhibit its activity. However, there is need for novel and smaller Fascin1 inhibitors. The aim of this study was to assess the effect of compound G2 in colorectal cancer cell lines and compare it to migrastatin in in vitro and in vivo assays. Molecular modeling, actin-bundling, cell viability, inmunofluorescence, migration, and invasion assays were carried out in order to test anti-migratory and anti-invasive properties of compound G2. In addition, the in vivo effect of compound G2 was evaluated in a zebrafish model of invasion. HCT-116 cells exhibited the highest Fascin1 expression from eight tested colorectal cancer cell lines. Compound G2 showed important inhibitory effects on actin bundling, filopodia formation, migration, and invasion in different cell lines. Moreover, compound G2 treatment resulted in significant reduction of invasion of DLD-1 overexpressing Fascin1 and HCT-116 in zebrafish larvae xenografts; this effect being less evident in Fascin1 known-down HCT-116 cells. This study proves, for the first time, the in vitro and in vivo anti-tumoral activity of compound G2 on colorectal cancer cells and guides to design improved compound G2-based Fascin1 inhibitors. Key messages center dot Fascin is crucial for tumor invasion and metastasis and is overexpressed in bad prognostic tumors. center dot Several adverse tumors overexpress Fascin1 and lack targeted therapy. center dot Anti-fascin G2 is for the first time evaluated in colorectal carcinoma and compared with migrastatin. center dot Filopodia formation, migration activity, and invasion in vitro and in vivo assays were performed. center dot G2 blocks actin structures, migration, and invasion of colorectal cancer cells as fascin-dependent.Peer reviewe

The FDA-Approved Antiviral Raltegravir Inhibits Fascin1-Dependent Invasion of Colorectal Tumor Cells In Vitro and In Vivo

Colorectal cancer (CRC) is the third leading cause of cancer-related deaths worldwide. Serrated adenocarcinoma (SAC) has been recently recognized by the WHO as a histological CRC with bad prognosis. Consistent with previous evidence, our group identified Fascin1 as a protein directly related to the invasiveness of tumor cells, overexpressed and positively correlated with worse survival in various carcinomas, including SAC. Therefore, Fascin1 has emerged as an ideal target for cancer treatment. In the present study, virtual screening has been carried out from a library of 9591 compounds, thus identifying the FDA-approved anti-retroviral raltegravir (RAL) as a potential Fascin1 blocker. In vitro and in vivo results show that RAL exhibits Fascin1-binding activity and Fascin1-dependent anti-invasive and anti-metastatic properties against CRC cells both in vitro and in vivo

New role of the antidepressant imipramine as a Fascin1 inhibitor in colorectal cancer cells

Colorectal cancer: Antitumor antidepressant The antidepressant drug imipramine can block the activity of a protein that contributes to the progression of certain aggressive tumors. Serrated adenocarcinoma (SAC) is a form of colorectal cancer with a poor prognosis. A key factor in SAC development is the overexpression of the protein fascin1, which promotes the formation of structures that help cancer cells move around, thereby leading to metastasis. Pablo Conesa-Zamora at Santa Lucia University Hospital in Cartagena, Horacio Perez-Sanchez at the Universidad Catolica de Murcia in Guadalupe, Spain, and coworkers demonstrated that imipramine shows promise in binding to fascin1 and blocking its activity. The team analyzed over 9500 compounds as potential fascin1 blockers, identifying imipramine as a possible option. In tests on human tissues and in vivo studies using zebrafish, the drug reduced cancer invasion and metastasis. Serrated adenocarcinoma (SAC) is more invasive, has worse outcomes than conventional colorectal carcinoma (CRC), and is characterized by frequent resistance to anti-epidermal growth factor receptor (EGFR) and overexpression of fascin1, a key protein in actin bundling that plays a causative role in tumor invasion and is overexpressed in different cancer types with poor prognosis. In silico screening of 9591 compounds, including 2037 approved by the Food and Drug Administration (FDA), was performed, and selected compounds were analyzed for their fascin1 binding affinity by differential scanning fluorescence. The results were compared with migrastatin as a typical fascin1 inhibitor. In silico screening and differential scanning fluorescence yielded the FDA-approved antidepressant imipramine as the most evident potential fascin1 blocker. Biophysical and different in vitro actin-bundling assays confirm this activity. Subsequent assays investigating lamellipodia formation and migration and invasion of colorectal cancer cells in vitro using 3D human tissue demonstrated anti-fascin1 and anti-invasive activities of imipramine. Furthermore, expression profiling suggests the activity of imipramine on the actin cytoskeleton. Moreover, in vivo studies using a zebrafish invasion model showed that imipramine is tolerated, its anti-invasive and antimetastatic activities are dose-dependent, and it is associated with both constitutive and induced fascin1 expression. This is the first study that demonstrates an antitumoral role of imipramine as a fascin1 inhibitor and constitutes a foundation for a molecular targeted therapy for SAC and other fascin1-overexpressing tumors.Peer reviewe

Admissions for hypoglycaemia after 35 weeks of gestation:perinatal predictors of cost of stay

Multivariate analysis of the clinicopathological and molecular factors associated with FOXD2 methylation (A); DIO3 and FOXD2 mRNA expression in hMSI-H tumoural and normal specimens (B) and external validation using TCGA database showing that MSI-H expressed less FOXD2 than MSI-L/MSS colon carcinomas (C)