Protein pathway activation mapping of colorectal metastatic progression reveals metastasis-specific network alterations

The mechanism by which tissue microecology influences invasion and metastasis is largely unknown. Recent studies have indicated differences in the molecular architecture of the metastatic lesion compared to the primary tumor, however, systemic analysis of the alterations within the activated protein signaling network has not been described. Using laser capture microdissection, protein microarray technology, and a unique specimen collection of 34 matched primary colorectal cancers (CRC) and synchronous hepatic metastasis, the quantitative measurement of the total and activated/phosphorylated levels of 86 key signaling proteins was performed. Activation of the EGFR-PDGFR-cKIT network, in addition to PI3K/AKT pathway, was found uniquely activated in the hepatic metastatic lesions compared to the matched primary tumors. If validated in larger study sets, these findings may have potential clinical relevance since many of these activated signaling proteins are current targets for molecularly targeted therapeutics. Thus, these findings could lead to liver metastasis specific molecular therapies for CRC. \uc2\ua9 2012 Springer Science+Business Media Dordrecht

Colorectal Cancer Stage at Diagnosis Before vs During the COVID-19 Pandemic in Italy

IMPORTANCE Delays in screening programs and the reluctance of patients to seek medical attention because of the outbreak of SARS-CoV-2 could be associated with the risk of more advanced colorectal cancers at diagnosis. OBJECTIVE To evaluate whether the SARS-CoV-2 pandemic was associated with more advanced oncologic stage and change in clinical presentation for patients with colorectal cancer. DESIGN, SETTING, AND PARTICIPANTS This retrospective, multicenter cohort study included all 17 938 adult patients who underwent surgery for colorectal cancer from March 1, 2020, to December 31, 2021 (pandemic period), and from January 1, 2018, to February 29, 2020 (prepandemic period), in 81 participating centers in Italy, including tertiary centers and community hospitals. Follow-up was 30 days from surgery. EXPOSURES Any type of surgical procedure for colorectal cancer, including explorative surgery, palliative procedures, and atypical or segmental resections. MAIN OUTCOMES AND MEASURES The primary outcome was advanced stage of colorectal cancer at diagnosis. Secondary outcomes were distant metastasis, T4 stage, aggressive biology (defined as cancer with at least 1 of the following characteristics: signet ring cells, mucinous tumor, budding, lymphovascular invasion, perineural invasion, and lymphangitis), stenotic lesion, emergency surgery, and palliative surgery. The independent association between the pandemic period and the outcomes was assessed using multivariate random-effects logistic regression, with hospital as the cluster variable. RESULTS A total of 17 938 patients (10 007 men [55.8%]; mean [SD] age, 70.6 [12.2] years) underwent surgery for colorectal cancer: 7796 (43.5%) during the pandemic period and 10 142 (56.5%) during the prepandemic period. Logistic regression indicated that the pandemic period was significantly associated with an increased rate of advanced-stage colorectal cancer (odds ratio [OR], 1.07; 95%CI, 1.01-1.13; P = .03), aggressive biology (OR, 1.32; 95%CI, 1.15-1.53; P < .001), and stenotic lesions (OR, 1.15; 95%CI, 1.01-1.31; P = .03). CONCLUSIONS AND RELEVANCE This cohort study suggests a significant association between the SARS-CoV-2 pandemic and the risk of a more advanced oncologic stage at diagnosis among patients undergoing surgery for colorectal cancer and might indicate a potential reduction of survival for these patients

Applicazione del Reverse Phase Protein Array per l'analisi del profilo fosfoprotemico nelle metastasi epatiche da carcinoma del colon-retto e identificazione di nuovi bersagli terapeutici

Background Hepatic metastases from colorectal cancer are still characterized by a poor prognosis as only 20% of patients can undergo surgical resection and in the other 80% chemotherapy achieves short results with a 5-years survival lower than 5%. Protein-kinases play a crucial role in determinating alterations of intracellular signal transmission that are basis for neoplastic and metastatic process. Reverse phase protein array is a technique that allow simultaneous analysis of the phosphorilation state from different specimens and consequently the activation of protein-kinases. Aim of the study Our purpose is to compare the phosphoproteomic profile expression of a kinases group involved in intracellular signal transmission of sybchronous liver metastases from colorectal cancer with the matched primary tumors and with metachronous lung metastases, to evaluate if there are a common phenotype for every type of metastases from colorectal cancer and a specific phenotype for liver metastases. Materials and methods Using reverse phase protein array we analyzed the activity of 87 protein-kinases involved in multiple intracellular signalling pathways of 34 specimens from patients with colorectal cancer and synchronous liver metastases and of 16 specimens from patients with metachronous colorectal cancer lung metastases. Results The levels of 38 out of 87 endpoints analyzed were significantly altered between the primary tumors and the liver metastases. The majority of these are components of only two major signaling pathways: these include the PI3 Kinase/AKT/mTOR prosurvival pathway and the growth factor receptor signaling pathways (which also feed into PI3K signaling). We also observed an increase in phosphorylation of PDGFRb (Y751) (p value = 0.0181), and cKit Y703 (p value = 0.005) in the liver metastases as compared to the primary tumors: both these growth factor receptors also signal through the PI3K/AKT pathway. To confirm this finding, Western blot analysis of AKT (S473) and c-Kit (Y703) was performed on 5 of the cases using a separately obtained cell population obtained by LCM: the results show that the elevations observed by RPPA are accurate. Comparing the liver metastases to the matched primary colorectal cancver tumor and the unmatched pulmonary metastasis phosphorylation of Pyk2 (Y402) and VEGFR (Y996) were found to be higher and phosphorylation of PKCzeta/lambda (T403/410) and BAD (S136) lower in the liver metastases (p <0.05). Conclusions Our results highlight that: 1- There is a common metastatic phenotype in neoplastic cells which is not related to target organ, the PI3 Kinase/AKT/mTOR prosurvival pathway. 2- There is also a phenotype specific for the liver metastases that include the overexpression of VEGFR2 (Y996) e Pyk2 (Y402) in the liver metastases. Moreover these results suggest some new targets for the therapeutic strategy of colorectal metastases: an inhibitor of PI3K-AKT, in association with drugs as Gleevec (Imatinib mesilate) that inhibit PDGFR receptors and c-KIT, might be used as for colorectal metastases. More selectively, for the treatment of liver metastases, an association of Bevacizumab (VEGFR inhibitor) with a Pyk2 inhibitor could be useful and effective.Introduzione La presenza di metastasi epatiche da carcinoma del colon-retto è contraddistinta da una prognosi infausta, poiché solo il 20% dei pazienti può essere sottoposto a resezione chirurgica e nella restante parte dei casi i risultati della chemioterapia sono scarsi, con sopravvivenza a 5 anni inferiore al 5%. Le protein-chinasi hanno un ruolo decisivo nel determinare le alterazioni della trasmissione del segnale intracellulare che caratterizzano sia il processo neoplastico che quello metastatico. Il reverse phase protein array è una metodica che permette di analizzare contemporaneamente in più campioni lo stato di fosforilazione e quindi di attivazione di numerose protein-chinasi. Scopo dello studio Scopo del progetto di ricerca è il confronto del profilo d’espressione fosfoproteomico di un gruppo di protein-chinasi implicate nella trasmissione del segnale intracellulare nel tumore del colon-retto primitivo e nelle rispettive metastasi epatiche e polmonari, per verificare se esiste un profilo d’espressione fosfoproteomico comune per le metastasi, inattivo nel tumore primitivo e parallelamente se esiste un profilo d’espressione specifico per le metastasi epatiche. Materiali e metodi Usando la tecnica reverse phase protein array, abbiamo analizzato l’attività di 87 protein-chinasi coinvolte in diverse vie di trasduzione del segnale intracellulare di 34 campioni provenienti da pazienti con adenocarcinoma del colon-retto con metastasi epatiche sincrone e di 16 campioni di metastasi polmonari metacrone da adenocarcinoma del colon-retto. Risultati I livelli di 38 protein chinasi sono risutati significativamente alterati nelle metastasi epatiche risptto ai rispettivi tumori primitivi. La maggior parte di queste protein-chinasi appartengono a due pathways di trasmissione del segnale intracellulare: il pathway PI3 kinase/AKT/mTOR e il pathway che regola l’apoptosi legata ai recettori per i growth factors, che prevede comunque l'attivazione di PI3 kinase/AKT/mTOR. Un’ulteriore conferma dell’importanza di tale via di segnale è data dall’iperattivazione nelle metastasi epatiche di 2 recettori tirosin-chinasici connessi con il pathway PI3 kinase/AKT/mTOR: PDGFRb Y751 (p = 0.0181) e cKit Y703 (p = 0.005), recettori invece ipofosforilati nel tumore primitivo. Questi risultati sono stati confermati mediante analisi con Western Blot dell’espressione di pAKT e di p-cKit su 5 casi. Confrontando le metastasi epatiche con le metastasi polmonari il livello di fosforilazione di 4 protein-chinasi è risultato alterato solo nelle metastasi epatiche: più precisamente la fosforilazione di Pyk2 (Y402) e VEGFR (Y996) è aumentata, mentre quella di PKCzeta/lambda (T403/410) e BAD (S136) è diminuita nelle metastasi epatiche rispetto alle metastasi polmonari ed al tumore primitivo (p < 0.05). Conclusioni: Questi risultati ci hanno portato alle seguenti considerazioni: 1- Esiste un pathway intracellulare iperattivato sia nelle metastasi epatiche che polmonari, rispetto al tumore primitivo, rappresentato dalla via mediata da PI3K-AKT 2- La sovraespressione di VEGFR2 (Y996) e Pyk2 (Y402) nelle metastasi epatiche ha permesso di delineare un secondo profilo fosfoproteomico specifico delle metastasi epatiche. Questi risultati hanno inoltre evidenziato nuovi possibili bersagli terapeutici per una terapia personalizzata con inibitori delle protein chinasi: un inibitore di PI3K-AKT assieme ad un inibitore di PDGFRb e c-Kit (Imatinib mesilato) per inibire il profilo fosfoproteomico comune alle metastasi epatiche e polmonari; un inibitore della via Pyk2-FAK (PF-562,271-01 Pfizer, Groton, Conn) per inibire il profilo fosfoproteomico tipico delle metastasi epatiche

Celiomesenteric Trunk Aneurysm: A Case Report

Abstract: A celiomesenteric trunk (CMT) is an extremely rare anatomic variant that consists of celiac and superior mesenteric arteries having a common origin from the aorta. CMT accounts for less than 1% of all splanchnic artery anomalies. Aneurysm involving a CMT is an even rarer vascular abnormality, and, to our knowledge, only eight cases of CMT aneurysm have been reported in literature. We describe a case of the incidental finding of CMT aneurysm in an asymptomatic patient. It was found after dorsolumbar column radiography and successive computed tomography and arteriography confirmed the diagnosis. Even if asymptomatic, we decided to repair it surgically with aneurysmectomy and suture of the neck due to risk of rupture

Primary great saphenous vein leiomyosarcoma: Report of a case

Abstract: Leiomyosarcomas rarely arise in primary veins, especially the great saphenous vein. We have found only 20 case reports of leiomyosarcoma arising in the great saphenous vein, most of which manifested as nonspecific symptoms of advanced disease, such as a palpable mass, swelling, and back or abdominal pain. We report the case of greater saphenous vein leiomyosarcoma diagnosed in a 48-year-old man with a 4-month history of an inguinal mass. Ultrasonography and computed tomography showed a 6-cm mass attached to the right superficial femoral vein. Fine-needle aspiration biopsy confirmed that it was a vascular sarcoma. At the time of surgery there was no evidence of distant metastasis; therefore, we removed the tumor en bloc along with the sartorius muscle, inguinal lymph nodes, and 10 cm of the common femoral vein, and replaced the femoral vein with a polytetrafluoroethylene graft. A pathological examination revealed poorly differentiated leiomyosarcoma of the great saphenous vein, involving the deep femoral vein, without lymph node involvement. During follow-up, a thrombosis of the prosthesis developed, followed by proximal stenosis, which was treated successfully with percutaneous transluminal angioplasty. The patient was found to have lung metastases 25 months after surgery and he died about 5 months later

Gene expression profile of primary gastric cancer: Towards the prediction of lymph node status

Background: The identification of gastric tumors associated with a higher risk of lymph node metastasis could help surgeons select patients who may benefit from extended lymph node dissection. The aim of this study was to screen the genome in the search of primary gastric cancer gene expression profiles that might predict lymph node status. Methods: The gene expression profile was evaluated in frozen tumor samples obtained from 32 patients with primary gastric adenocarcinomas. The array consisted of a duplicated spot panel of 5,541 human genes. To classify node-positive (N+) and node-negative (N-) cases, a logistic regression model was fitted optimizing the Akaike Information Criteria after a stepwise gene selection. The accuracy was evaluated by means of leave-one-out cross validation. Results: All patients underwent radical gastrectomy and extended lymphadenectomy. Of all the cases, 21 were N+ and 11 demonstrated no lymph node involvement (N-). After quality filtering, the analysis of variance selected a set of 136 genes potentially correlated with nodal involvement (P value <.05). Of these 136 genes, 5 were differentially expressed (adjusted P value <.05). After a stepwise gene selection, only three genes (Bik, aurora kinase B, eIF5A2) were retained in the logistic model, which could correctly predict lymph node status in 30 of 32 cases. Conclusions: If our findings were confirmed, the identified gene pattern might be used to tailor the extent of lymph node dissection on a single patient basis

Outcomes of Laparoscopic Surgery in Very Elderly Patients with Colorectal Cancer: A Survival Analysis and Comparative Study

(1) Background: Colorectal cancer (CRC) is a global health concern, particularly among the elderly population. This study aimed to assess the impact of laparoscopic surgery on CRC patients aged ≥80 years. (2) Methods: We conducted a retrospective analysis of prospectively collected data from consecutive CRC patients who underwent surgery at our institution between July 2018 and July 2023. The patients were categorized into three groups: those aged over 80 who underwent laparoscopic surgery (Group A), those aged over 80 who underwent open surgery (Group B), and those under 80 who underwent laparoscopic surgery (Group C). We examined various clinical and surgical parameters, including demographic data, medical history, surgical outcomes, and survival. (3) Results: Group A (N = 113) had shorter hospital stays than Group B (N = 23; p = 0.042), with no significant differences in complications or 30-day outcomes. Compared to Group C (N = 269), Group A had higher comorbidity indices (p p p p = 0.003). Laparotomic conversion was associated with obstructive neoplasms (p p p = 0.007 and p p p p = 0.136). (4) Conclusions: Laparoscopic surgery in very elderly CRC patients shows comparable oncological outcomes and surgical complications to younger populations. Survival benefits are influenced by age, comorbidities, and medical complications. Further prospective multicenter studies are needed in order to validate these findings

A Pilot Characterization of Human Lung NSCLC by Protein Pathway Activation Mapping

Abstract: Background: An understanding of the activated protein signaling architecture in non-small-cell lung cancer (NSCLC) is of critical importance to the development of new therapeutic approaches and identification of predictive and prognostic biomarkers for patient stratification. Methods: We used reverse-phase protein microarrays to map the activated protein signaling networks of 47 NSCLC tumors, 28 of which were node negative, which were subjected to tumor cellular enrichment using laser capture microdissection. The phosphorylation/cleavage levels of 111 key signaling proteins and total levels of 17 proteins were measured for broadscale signaling analysis. Results: Pathway activation mapping of NSCLC revealed distinct subgroups composed of epidermal growth factor receptor (ERBB1), v-erb-b2 erythroblastic leukemia viral oncogene homolog 2 (ERBB2), v-erb-b2 erythroblastic leukemia viral oncogene homolog 3 (ERBB3), v-erb-a erythroblastic leukemia viral oncogene homolog 4 (ERBB4), v-akt murine thymoma viral oncogene homolog 1-mammalian target of rapamycin (AKT-mTOR), protein kinase, AMP-activated, alpha 2 catalytic subunit (AMPK), and autophagy-related signaling, along with transforming growth factor-beta-signaling protein 1 (SMAD), insulin-line growth factor receptor (IGFR), rearranged during transfection proto-oncogene (RET), and activated CDC42-associated kinase (ACK) activation. Investigation of epidermal growth factor receptor (EGFR)-driven signaling identified a unique cohort of tumors with low EGFR protein expression yet high relative levels of phosphorylated EGFR and high EGFR total protein with low relative levels of phosphorylation. Last, mapping analysis of patients with NSCLC with N0 disease revealed a pilot pathway activation signature composed of linked epidermal growth factor receptor family (HER)-AMPK-AKT-mTOR signaling network along with focal adhesion kinase-LIM domain kinase-1 (FAK-LIMK) and janus kinase (JAK)-signal transducers and activators of transcription (STAT) pathways that correlated with short-term survival and aggressive disease. Conclusions: Functional protein pathway activation mapping of NSCLC reveals distinct activation subgroups that are underpinned by important therapeutic targets and that patients with early-stage node negative disease and poor prognosis may be identified by activation of defined, biochemically linked protein signaling events. Such findings, if confirmed in larger study sets, could help select and stratify patients for personalized targeted therapies

Circulating miR-182 is a biomarker of colorectal adenocarcinoma progression

MiR-182 expression was evaluated by qRT-PCR and in situ hybridization in 20 tubular adenomas, 50 colorectal carcinoma (CRC), and 40 CRC liver metastases. Control samples obtained from patients with irritable bowel syndrome, or tumor-matched normal colon mucosa were analyzed (n=50). MiR-182 expression increased progressively and significantly along with the colorectal carcinogenesis cascade, and in CRC liver metastases. The inverse relation between miR-182 and the expression of its target gene ENTPD5 was investigated by immunohistochemical analysis. We observed that normal colocytes featured a strong ENTPD5 cytoplasmic expression whereas a significantly and progressively lower expression was present along with dedifferentiation of the histologic phenotype. Plasma samples from 51 CRC patients and controls were tested for miR-182 expression. Plasma miR-182 concentrations were significantly higher in CRC patients than in healthy controls or patients with colon polyps at endoscopy. Moreover, miR-182 plasma levels were significantly reduced in post-operative samples after radical hepatic metastasectomy compared to preoperative samples. Our results strengthen the hypothesis of a central role of miR-182 dysregulation in colon mucosa transformation, demonstrate the concomitant progressive down-regulation of ENTPD5 levels during colon carcinogenesis, and indicate the potential of circulating miR-182 as blood based biomarker for screening and monitoring CRC during the follow-up