Unmutated RRAS2 emerges as a key oncogene in post-partum-associated triple negative breast cancer.

BACKGROUND: Breast cancer (BC) is the most common cancer in women, with triple negative BC (TNBC) accounting for 20% of cases. While early detection and targeted therapies have improved overall life expectancy, TNBC remains resistant to current treatments. Although parity reduces the lifetime risk of developing BC, pregnancy increases the risk of developing TNBC for years after childbirth. Although numerous gene mutations have been associated with BC, no single gene alteration has been identified as a universal driver. RRAS2 is a RAS-related GTPase rarely found mutated in cancer. METHODS: Conditional knock-in mice were generated to overexpress wild type human RRAS2 in mammary epithelial cells. A human sample cohort was analyzed by RT-qPCR to measure RRAS2 transcriptional expression and to determine the frequency of both a single-nucleotide polymorphism (SNP rs8570) in the 3'UTR region of RRAS2 and of genomic DNA amplification in tumoral and non-tumoral human BC samples. RESULTS: Here we show that overexpression of wild-type RRAS2 in mice is sufficient to develop TNBC in 100% of females in a pregnancy-dependent manner. In human BC, wild-type RRAS2 is overexpressed in 68% of tumors across grade, location, and molecular type, surpassing the prevalence of any previously implicated alteration. Still, RRAS2 overexpression is notably higher and more frequent in TNBC and young parous patients. The increased prevalence of the alternate C allele at the SNP position in tumor samples, along with frequent RRAS2 gene amplification in both tumors and blood of BC patients, suggests a cause-and-effect relationship between RRAS2 overexpression and breast cancer. CONCLUSIONS: Higher than normal expression of RRAS2 not bearing activating mutations is a key driver in the majority of breast cancers, especially those of the triple-negative type and those linked to pregnancy.

Identification of novel targets in adipose tissue involved in non-alcoholic fatty liver disease progression.

Obesity is a major risk factor for the development of Nonalcoholic fatty liver disease (NAFLD). We hypothesize that a dysfunctional subcutaneous white adipose tissue (scWAT) may lead to an accumulation of ectopic fat in the liver. Our aim was to investigate the molecular mechanisms involved in the causative role of scWAT in NALFD progression. We performed a RNA-sequencing analysis in a discovery cohort (n = 45) to identify genes in scWAT correlated with fatty liver index, a qualitative marker of liver steatosis. We then validated those targets in a second cohort (n = 47) of obese patients who had liver biopsies available. Finally, we obtained scWAT mesenchymal stem cells (MSCs) from 13 obese patients at different stages of NAFLD and established in vitro models of human MSC (hMSC)-derived adipocytes. We observed impaired adipogenesis in hMSC-derived adipocytes as liver steatosis increased, suggesting that an impaired adipogenic capacity is a critical event in the development of NAFLD. Four genes showed a differential expression pattern in both scWAT and hMSC-derived adipocytes, where their expression paralleled steatosis degree: SOCS3, DUSP1, SIK1, and GADD45B. We propose these genes as key players in NAFLD progression. They could eventually constitute potential new targets for future therapies against liver steatosis.

Radio-Pathological Correlations in Patients with Liver Metastases for Colorectal Cancer.

BACKGROUND: Colorectal cancer (CRC) is the most frequent gastrointestinal cancer. The liver is the organ most commonly affected by CRC metastases. Synchronous CRC liver metastases (CRCLM) are present in 15-25% at diagnosis, and metastases are confined to the liver in 70-80% of these cases. The aim of the present study was to investigate the existence of significant correlations between the pathological features and computed tomography scan morpho-densitometric findings. SUMMARY: A retrospective study of prospectively collected data has been performed; all patients underwent curative-intent hepatic resection from January 2004 to December 2012 and had histologically confirmed CRCLM. Key Messages: Thirty-four (57%) patients were males; the mean age was 64.4 (±10.2) years. Statistically significant differences have been found with the percentages of intra-tumoral fibrosis (p = 0.038) and necrosis (p = 0.007); the values of fibrosis are higher in the absence of a peri-lesional ring, while those of necrosis are higher in the presence of a peri-lesional ring.There was a correlation between the histopathological response to treatments and the global attenuation levels observed in the computed tomography scan of CRCLM. Furthermore, the presence of a radiologically evidenced peripheral ring was associated with the amount of viable tumor cells in the periphery of the tumor, and with responses predominated by necrosis. More studies are needed to clarify the radiological and histological correlation and to be able to better select patients who are going to undergo surgery.

Nuclear IGF-1R predicts chemotherapy and targeted therapy resistance in metastatic colorectal cancer.

BACKGROUND: Although chemotherapy is the cornerstone treatment for patients with metastatic colorectal cancer (mCRC), acquired chemoresistance is common and constitutes the main reason for treatment failure. Monoclonal antibodies against insulin-like growth factor-1 receptor (IGF-1R) have been tested in pre-treated mCRC patients, but results have been largely deceiving. METHODS: We analysed time to progression, overall survival, and the mutational status of RAS, BRAF and nuclear p-IGF-1R expression by immunohistochemistry, in 470 metastatic CRC patients. The effect of IGF-1R activation and distribution was also assessed using cellular models of CRC and RNAi for functional validation. RESULTS: Nuclear IGF-1R increased in metastatic tumours compared to paired untreated primary tumours, and significantly correlated with poor overall survival in mCRC patients. In vitro, chemo-resistant cell lines presented significantly higher levels of IGF-1R expression within the nuclear compartment, and PIAS3, a protein implicated also in the sumoylation process of intranuclear proteins, contributed to IGF-1R nuclear sequestration, highlighting the essential role of PIAS3 in this process. Intriguingly, we observed that ganitumab, an IGF-1R blocking-antibody used in several clinical trials, and dasatinib, an SRC inhibitor, increased the nuclear localisation of IGF-1R. CONCLUSIONS: Our study demonstrates that IGF-1R nuclear location might lead to chemotherapy and targeted agent resistance.

Unraveling Adipose Tissue Dysfunction: Molecular Mechanisms, Novel Biomarkers, and Therapeutic Targets for Liver Fat Deposition.

Adipose tissue (AT), once considered a mere fat storage organ, is now recognized as a dynamic and complex entity crucial for regulating human physiology, including metabolic processes, energy balance, and immune responses. It comprises mainly two types: white adipose tissue (WAT) for energy storage and brown adipose tissue (BAT) for thermogenesis, with beige adipocytes demonstrating the plasticity of these cells. WAT, beyond lipid storage, is involved in various metabolic activities, notably lipogenesis and lipolysis, critical for maintaining energy homeostasis. It also functions as an endocrine organ, secreting adipokines that influence metabolic, inflammatory, and immune processes. However, dysfunction in WAT, especially related to obesity, leads to metabolic disturbances, including the inability to properly store excess lipids, resulting in ectopic fat deposition in organs like the liver, contributing to non-alcoholic fatty liver disease (NAFLD). This narrative review delves into the multifaceted roles of WAT, its composition, metabolic functions, and the pathophysiology of WAT dysfunction. It also explores diagnostic approaches for adipose-related disorders, emphasizing the importance of accurately assessing AT distribution and understanding the complex relationships between fat compartments and metabolic health. Furthermore, it discusses various therapeutic strategies, including innovative therapeutics like adipose-derived mesenchymal stem cells (ADMSCs)-based treatments and gene therapy, highlighting the potential of precision medicine in targeting obesity and its associated complications.

Gene-expression signature of tumor recurrence in patients with stage II and III colon cancer treated with 5'fluoruracil-based adjuvant chemotherapy.

Although receiving adjuvant chemotherapy after radical surgery, a disappointing proportion of patients with colorectal cancer will develop tumor recurrence. Probability of relapse is currently predicted from pathological staging, there being a need for additional markers to further select high-risk patients. This study was aimed to identify a gene-expression signature to predict tumor recurrence in patients with Stages II and III colon cancer treated with 5'fluoruracil (5FU)-based adjuvant chemotherapy. Two-hundred and twenty-eight patients diagnosed with Stages II-III colon cancer and treated with surgical resection and 5FU-based adjuvant chemotherapy were included. RNA was extracted from formalin-fixed, paraffin-embedded tissue samples and expression of 27 selected candidate genes was analyzed by RT-qPCR. A tumor recurrence predicting model, including clinico-pathological variables and gene-expression profiling, was developed by Cox regression analysis and validated by bootstrapping. The regression analysis identified tumor stage and S100A2 and S100A10 gene expression as independently associated with tumor recurrence. The risk score derived from this model was able to discriminate two groups with a highly significant different probability of tumor recurrence (HR, 2.75; 95%CI, 1.71-4.39; p = 0.0001), which it was maintained when patients were stratified according to tumor stage. The algorithm was also able to distinguish two groups with different overall survival (HR, 2.68; 95%CI, 1.12-6.42; p = 0.03). Identification of a new gene-expression signature associated with a high probability of tumor recurrence in patients with Stages II and III colon cancer receiving adjuvant 5FU-based chemotherapy, and its combination in a robust, easy-to-use and reliable algorithm may contribute to tailor treatment and surveillance strategies.

The Validity of the Invasive Tests for Helicobacter Pylori Diagnosis is Unequally Affected by the Consumption of Antibiotics or Pump Inhibitors. Test Performance under Real-World Conditions.

BACKGROUND AND AIMS: Patients undergoing upper endoscopy have often used proton pump inhibitors (PPI) and/or antibiotics (ABx) recently. Both drugs have been associated with a poorer yield of the Helicobacter pylori (H. pylori) diagnostic tests. The aim was to assess the accuracy of the polymerase chain reaction test (qPCR), histological exam (HE) and ultra-fast urease test (UFUT) for H. pylori detection in patients that recently used PPI or ABx. METHODS: Prospective study recruiting 206 patients who underwent upper endoscopy and gastric biopsies. Demographics and use of PPI/ABx were obtained. Sensibility (Sn), specificity (Sp), predictive value (PV), likelihood ratio (LR) and PABAK concordance index, were calculated, considering as the gold standard the positivity of 2 out of 3 analyzed tests. A global analysis and another one based on the PPI/ABx intake were performed. RESULTS: 48.5% of patients used PPI and 12.8% ABx within the 2 and 4 weeks prior to endoscopy, respectively. The UFUT was positive in 13.1% of patients, HE in 34% and qPCR in 35.9%. UFUT achieved lower Sn (37%) than HE (98%) and qPCR (98%) (p<0.001) overall. ABx were associated with lower Sn in HE (p=0.04) and lower Sp in qPCR (p=0.03). PPI did not associate with a significant drop in Sn and Sp. The concordance between HE and qPCR was 0.83 (95%CI: 0.73-0.89). CONCLUSIONS: Under real world conditions, the accuracy and concordance of HE and qPCR to diagnose H. pylori were excellent, but UFUT achieved unsatisfactory outcomes. The intake of ABx was associated with the worse performance, fundamentally for HE. The PPI did not reduce the tests' yield significantly.

Sperm Tail Defects and Abnormal Testicular Blood Flow in a Beagle Dog: A Case Report.

A 5-year-old male Beagle dog produced ejaculates with a high percentage of spermatozoa with abnormal morphology, especially sperm tail defects. Although libido and semen volume were normal, ejaculates showed asthenospermia, oligozoospermia, and teratozoospermia. The spermatozoa exhibited morphologic defects affecting the flagellum, mainly coiled tails with or without macrocephalia (33.5 ± 2.1%), bent tails (18.3 ± 3.4%), and proximal cytoplasmic droplets (6.7 ± 2.8%). The peripheral plasma testosterone level was 2.76 ± 0.21 ng/mL. The resistive index and the pulsatility index from marginal and intratesticular vessels measured by Doppler ultrasound showed higher values in the right testicle than in the left testicle. Histologic evaluation revealed focal reduction in the number of germ cells and sperm in the seminiferous tubules in the right testicle. This is the first report that describes simultaneously the presence of sperm tail defects in the ejaculate and changes in the blood flow of testicular vessels in the dog.

Phosphorylated-insulin growth factor I receptor (p-IGF1R) and metalloproteinase-3 (MMP3) expression in advanced gastrointestinal stromal tumors (GIST). A GEIS 19 study.

BACKGROUND: Most GISTs have mutations in KIT or PDGFRA. Patients with advanced GIST with KIT exon 9, PDGFRA mutation or WT for KIT and PDGFRA have a worse progression-free survival (PFS) compared to patients with KIT exon 11 mutated tumors. We evaluated the immunohistochemical (IHC) expression of p-IGF1R (Y1316) and MMP3 as predictors of PFS or overall survival (OS). METHODS: Ninety-two advanced GIST patients included in GEIS-16 study with KIT and PDGFRA mutational information were examined for p-IGF1R (Y1316) and MMP3 expression in a tissue micro-array. To study activation of the IGF1R system, we have used an antibody (anti-pY1316) that specifically recognizes the active phosphorylated form of the IGF1R. DNA was extracted from paraffin-embedded tissues and intronic PCR primers were used to amplify exons 9, 11, 13 and 17 of KIT, 12 and 18 of PDGFRA. Bidirectional sequencing with specific primers was performed on a ABI3100 sequencer using the Big Dye Terminator v3.1 kit. Multivariate model was built using a stepwise automated variable selection approach with criterion to enter the variable in the model of p < 0.10 and criterion to keep the variable in the model of p < 0.05. PFS was computed as the date of imatinib initiation to progression or death. Overall survival was defined as the time from imatinib initiation to death. RESULTS: Phospho-IGF1R was expressed only in 9 % (2/22) of cases without KIT mutation. MMP3 expression was detected in 2/5 patients (40 %) with PDGFRA mutation, 1/16 patients (6 %) with WT genotype and 7/71 patients (10 %) of KIT mutant patients. At univariate analysis KIT exon 11/13 mutation had better PFS than patients with exon 9 mutation, PDGFRA mutation or WT genotype (p = 0.021; HR: 0.46; 95 %CI (0.28-0.76). Less than 24 months disease free-interval (HR 24.2, 95 % CI 10.5-55.8), poor performance status (PS) (HR 6.3, 95 % CI 2.5-15.9), extension of disease; >1 organ (HR 1.89; 95 % CI 1.03-3.4) and genotype analysis (HR 0.57, 95 % CI 0.37-0.97) but not immunophenotype analysis (HR 1.53; 95 % CI 0.76-3.06) were the strongest prognostic factors for PFS in the multivariate analysis. CONCLUSIONS: Our results do not support p-IGF-1R and MMP3 evaluation in non-selected GIST patients but evaluation of this immunophenotype in WT and mutant PDGFR mutation in larger group of GIST patients, deserve merits.

RAC1b overexpression correlates with poor prognosis in KRAS/BRAF WT metastatic colorectal cancer patients treated with first-line FOLFOX/XELOX chemotherapy.

INTRODUCTION: Chemotherapy is the principal treatment in metastatic colorectal cancer (mCRC) patients. RAC1b, a RAC1 spliced variant, is over-expressed in colorectal cancer (CRC), and impairs apoptosis by activation of nuclear-factor-KB. Since RAC1b has been associated with the BRAF(V600E) mutation, associated with poor prognosis in CRC, we evaluated the role of RAC1b expression as a predictor of chemotherapy efficacy in mCRC. METHODS: We analysed KRAS and BRAF mutation, microsatellite instability and RAC1b expression in 157 mCRC patients treated with FOLFOX/XELOX in first-line therapy. RESULTS: KRAS mutations were detected in 46 patients (34%), 10 patients were BRAF mutant (7%) and 79 were WT for both, KRAS and BRAF (59%). RAC1b overexpression was found in 30 patients (19%). In the multivariate analysis, BRAF mutational status was a poor prognostic factor for overall survival (OS); hazard ratio (HR), 2.78 (95% confidence interval (CI), 1.35-5.72; p=0.0057). RAC1b overexpression was a poor survival factor for OS (HR, 2.35; 95% CI, 1.2-4.59; p=0.01) and progression-free survival (PFS) (HR, 2.4; 95% CI, 1.2-4.78; p=0.01) in KRAS/BRAF WT mCRC patients. CONCLUSIONS: RAC1b overexpression constitutes a marker of poor prognosis in KRAS/BRAF WT mCRC patients treated with first-line FOLFOX/XELOX therapy.

Co-expression of matrix metalloproteinase-7 (MMP-7) and phosphorylated insulin growth factor receptor I (pIGF-1R) correlates with poor prognosis in patients with wild-type KRAS treated with cetuximab or panitumumab: a GEMCAD study.

BACKGROUND: By transactivacion, phosphorylated insulin growth factor receptor I (IGF-1R) can activate epidermal growth factor receptor (EGFR). MMP-7, produced by colorectal cancer cells, also can activate IGF-1R by degrading IGFBP-3 and releasing IGF-I. METHODS: A cohort of patients (pts) with advanced colorectal cancer (CRC), under second- or third-line treatment with cetuximab or panitumumab, was tested using immunohistochemistry for expression of the activated form of IGF-1R (p-IGF-1R) and MMP-7. KRAS and BRAF mutation status was determined by sequencing and allelic discrimination analysis, respectively. Analyses were performed in primary CRC tumor samples or metastases, and the association of immunohistochemistry findings, mutational results, and treatment outcomes was investigated in both univariate and multivariate analyses. RESULTS: Expression of activated IGF-1R and MMP-7 was observed in 51 and 49% of pts, respectively. Co-expression of MMP-7 and pIGF-1R (double positivity, DP) was observed in 28 pts (25%). There was no association between KRAS or BRAF mutational status and DP (p=0.52). Pts with DP responded more poorly to first-line chemotherapy (p=0.005) and to anti-EGFR treatment (p=0.01) than non-DP pts. In wild type (WT) KRAS pts, those with DP have poorer PFS (2.7 months vs. 3.5m, p=0.036; HR 1.98, 95% CI 1.05-3.75) and OS (6.4 months vs. 8.6 m, p=0.010; HR 2.33, 95%CI 1.23-4.43) in the adjusted multivariate analysis. CONCLUSIONS: Our study suggests that concomitant expression of MMP-7 and activation of p-IGF-1R (DP) correlates with poor prognosis in WT KRAS pts treated with anti-EGFR.

Sarcoma histiocítico en intestino delgado: estudio y discusión de un caso con reordenamiento clonal linfoide B / Lymphoid B clonal rearrangement in histiocytic sarcoma of the small intestine. A case report

El sarcoma histiocítico es un tumor maligno infrecuente, de patogenia incierta, caracterizado por una proliferación de células neoplásicas con morfología e inmunofenotipo de los histiocitos tisulares maduros. Aunque existen pocas referencias bibliográficas, afecta a pacientes de ambos sexos, de amplio rango de edad y afectación nodal y/o extranodal. Se ha señalado la existencia en muchos casos de una estrecha relación entre esta entidad y varias neoplasias hematolinfoides en un mismo paciente, lo que ha dado lugar a múltiples estudios para esclarecer su etiología y su patogenia. Su curso suele ser rápidamente progresivo y no se conoce una pauta terapéutica eficaz. Actualmente su diagnóstico sigue siendo por exclusión. Presentamos el caso de un paciente de 82años con perforación de un asa yeyunal con una tumoración maligna que infiltra transmuralmente la pared y solo expresa CD68, CD45RO, CD163, lisozima y vimentina, junto con un reordenamiento clonal linfoideB con escasa amplificación (AU)

Histiocytic sarcoma is a rare malignant tumour of unknown pathogenesis characterized by proliferation of neoplastic cells morphologically and immunophenotypically similar to mature tissue histiocytes. There are only a few reported cases, but they have been described in both males and females of all ages and with nodal and extranodal involvement. A close relationship has often been observed with other haematolymphoid neoplasms which might provide clues to its etiology and pathogenesis. To date, diagnosis is by exclusion of other entities. The clinical course usually progresses rapidly and an effective therapeutic regime has not yet been established. We report a case in an 82year old male who had suffered a perforation of the jejunal loop and was found to have a malignant tumour infiltrating the wall of the small bowel. The tumour was positive only for CD68, CD45RO, CD163, lysozyme and vimentin and showed a Blymphoid clonal rearrangement with little amplification (AU)

Recomendaciones para el estudio patológico de las metástasis hepáticas de cáncer colorrectal tras neoadyuvancia. Consenso Nacional de la Sociedad Española de Anatomía Patológica / Recommendations for the study of liver metastases of colorectal cancer after neoadjuvant therapy. National Consensus of the Spanish Society of Pathology (SEAP)

Las resecciones hepáticas por metástasis de cáncer colorrectal (CCR) son una pieza quirúrgica frecuente en muchos servicios de anatomía patológica. Si al aumento de la incidencia de CCR añadimos otros factores como la frecuencia de metástasis hepáticas sincrónicas o metacrónicas, la ampliación en los criterios quirúrgicos de resecabilidad y el tratamiento neoadyuvante que facilita la resecabilidad, nos encontramos con una patología en aumento. El estudio anatomopatológico de estas piezas quirúrgicas se ha modificado y ha aumentado su complejidad debido a que se deben valorar nuevos datos histológicos como son los cambios potenciales producidos por el tratamiento neoadyuvante quimioterápico en el hígado no tumoral, y en el tumor el grado de regresión tumoral patológico, por su valor pronóstico. Teniendo en cuenta estos antecedentes, un grupo de patólogos se propuso revisar su papel en el diagnóstico y pronóstico de los pacientes con metástasis hepáticas de CCR con el objetivo de elaborar unas recomendaciones prácticas de procedimiento. De esta revisión se han obtenido unas directrices que podrían ser adaptadas por los distintos departamentos de patología con el fin de unificar procedimientos y obtener diagnósticos comparables. En este trabajo se exponen los resultados de este consenso (AU)

Liver resections for colorectal cancer (CRC) metastasis are common in most pathology departments. In addition, the frequency of liver resections for CRC specimens has increased due to an increased incidence of CRC the frequency of synchronous or metachronous liver metastases, the use of neoadjuvant therapy, and the increased surgical criteria of resectability. The pathological study of the specimens should include new histological data, i.e.: changes caused by therapy, both in the tumour and in the liver parenchyma, such as the pathological tumour regression grade, and the histologic degree of liver damage by the therapy, because of its prognostic value. On this setting, a group of pathologists has elaborated a guideline proposal, in order to obtain a more uniform procedure and diagnosis of CRC liver metastasis specimens. The aim was to give useful recommendations in order to obtain homogeneous and comparable pathologic reports among different pathology departments. The results of this consensus are presented in this paper (AU)

Evaluación de dispositivos automatizados para diagnóstico citológico en la prevención del cáncer de cérvix / Assessment of cytological automated devices in cervical cancer prevention

Antecedentes. La eficacia de las campañas de cribado para prevenir el cáncer de cérvix han demostrado su eficacia, especialmente en los paises desarrollados, donde su incidencia ha bajado notablemente. Planificar dichas campañas exige una infraestructura organizada en varios niveles, donde los servicios de Anatomía Patológica soportan la responsabilidad del diagnóstico. La comple-jidad del proceso mental que lleva a un diagnóstico citológico ha impedido hasta ahora el desarrollo de dispositivos automáticos que alivien la presión laboral que ejerce la enorme casuística generada y la rutinización. La moderna tecnología informática y de análisis de imágenes permiten hoy ya abor-dar este problema, tanto desde el punto de vista diagnóstico como para una estrategia de control de calidad en evitación de falsos negativos. Sobre estas bases nos proponemos evaluarla contribución que los dispositivos automáticos disponibles aportarían a la detección y control de calidad de las campañas de prevención del cáncer de cervix. Métodos. Revisión de la literatura. Resultados. En el momento de finalizar esta evaluación hemos detectado tres dispositivos cuya tecnología y grado de desarrollo difiere: AutoPap Primary Screening System (TriPath Imaging), ThinPrep Imaging System (Cytyc) e InPath (Molecular Diagnostics). Conclusiones. Los dispositivos automáticos de que se dispone actualmente se encuentran insuficientemente desarrollados (sólo AutoPap Imaging System cuenta con la aprobación de la FDA). Los recursos económicos y humanos deberían, al menos en el momento presente, dirigirse a fomentar institucionalmente el seguimiento de los programas de cri-bado y a apoyar organizativamente dichas campañas, más que a financiar equipos automáticos de diagnóstico. No obstante no debería dejarse de apoyar el desarrollo de estos dispositivos ya que, a medio plazo, con el previsible incremento de la población controlada y alcanzando un razonable equi-librio coste-eficacia, es muy probable que sea necesario apoyarse en esta tecnología. (AU)