Automated Prediction of the Response to Neoadjuvant Chemoradiotherapy in Patients Affected by Rectal Cancer

Simple Summary Colorectal cancer is the second most malignant tumor per number of deaths after lung cancer and the third per number of new cases after breast and lung cancer. The correct and rapid identification (i.e., segmentation of the cancer regions) is a fundamental task for correct patient diagnosis. In this study, we propose a novel automated pipeline for the segmentation of MRI scans of patients with LARC in order to predict the response to nCRT using radiomic features. This study involved the retrospective analysis of T-2-weighted MRI scans of 43 patients affected by LARC. The segmentation of tumor areas was on par or better than the state-of-the-art results, but required smaller sample sizes. The analysis of radiomic features allowed us to predict the TRG score, which agreed with the state-of-the-art results. Background: Rectal cancer is a malignant neoplasm of the large intestine resulting from the uncontrolled proliferation of the rectal tract. Predicting the pathologic response of neoadjuvant chemoradiotherapy at an MRI primary staging scan in patients affected by locally advanced rectal cancer (LARC) could lead to significant improvement in the survival and quality of life of the patients. In this study, the possibility of automatizing this estimation from a primary staging MRI scan, using a fully automated artificial intelligence-based model for the segmentation and consequent characterization of the tumor areas using radiomic features was evaluated. The TRG score was used to evaluate the clinical outcome. Methods: Forty-three patients under treatment in the IRCCS Sant'Orsola-Malpighi Polyclinic were retrospectively selected for the study; a U-Net model was trained for the automated segmentation of the tumor areas; the radiomic features were collected and used to predict the tumor regression grade (TRG) score. Results: The segmentation of tumor areas outperformed the state-of-the-art results in terms of the Dice score coefficient or was comparable to them but with the advantage of considering mucinous cases. Analysis of the radiomic features extracted from the lesion areas allowed us to predict the TRG score, with the results agreeing with the state-of-the-art results. Conclusions: The results obtained regarding TRG prediction using the proposed fully automated pipeline prove its possible usage as a viable decision support system for radiologists in clinical practice

The Heterogeneity of Skewness in T2W-Based Radiomics Predicts the Response to Neoadjuvant Chemoradiotherapy in Locally Advanced Rectal Cancer

Our study aimed to investigate whether radiomics on MRI sequences can differentiate responder (R) and non-responder (NR) patients based on the tumour regression grade (TRG) assigned after surgical resection in locally advanced rectal cancer (LARC) treated with neoadjuvant chemoradiotherapy (nCRT). Eighty-five patients undergoing primary staging with MRI were retrospectively evaluated, and 40 patients were finally selected. The ROIs were manually outlined in the tumour site on T2w sequences in the oblique-axial plane. Based on the TRG, patients were grouped as having either a complete or a partial response (TRG = (0,1), n = 15). NR patients had a minimal or poor nCRT response (TRG = (2,3), n = 25). Eighty-four local first-order radiomic features (RFs) were extracted from tumour ROIs. Only single RFs were investigated. Each feature was selected using univariate analysis guided by a one-tailed Wilcoxon rank-sum. ROC curve analysis was performed, using AUC computation and the Youden index (YI) for sensitivity and specificity. The RF measuring the heterogeneity of local skewness of T2w values from tumour ROIs differentiated Rs and NRs with a p-value ≈ 10−5; AUC = 0.90 (95%CI, 0.73–0.96); and YI = 0.68, corresponding to 80% sensitivity and 88% specificity. In conclusion, higher heterogeneity in skewness maps of the baseline tumour correlated with a greater benefit from nCR

Suppression of the HBP Function Increases Pancreatic Cancer Cell Sensitivity to a Pan-RAS Inhibitor

Pancreatic ductal adenocarcinoma (PDAC) is a leading cause of cancer-related death and the search for a resolutive therapy is still a challenge. Since KRAS is commonly mutated in PDAC and is one of the main drivers of PDAC progression, its inhibition should be a key strategy for treatment, especially considering the recent development of specific KRAS inhibitors. Nevertheless, the effects of KRAS inhibition can be increased through the co-inhibition of other nodes important for cancer development. One of them could be the hexosamine biosynthetic pathway (HBP), whose enhancement is considered fundamental for PDAC. Here, we demonstrate that PDAC cells expressing oncogenic KRAS, owing to an increase in the HBP flux, become strongly reliant on HBP for both proliferation and survival. In particular, upon treatment with two different compounds, 2-deoxyglucose and FR054, inhibiting both HBP and protein N-glycosylation, these cells undergo apoptosis significantly more than PDAC cells expressing wild-type KRAS. Importantly, we also show that the combined treatment between FR054 and the pan-RAS inhibitor BI-2852 has an additive negative effect on cell proliferation and survival by means of the suppression of both Akt activity and cyclin D1 expression. Thus, co-inhibition of HBP and oncogenic RAS may represent a novel therapy for PDAC patients

The Nutrient-Sensing Hexosamine Biosynthetic Pathway as the Hub of Cancer Metabolic Rewiring

Alterations in glucose and glutamine utilizing pathways and in fatty acid metabolism are currently considered the most significant and prevalent metabolic changes observed in almost all types of tumors. Glucose, glutamine and fatty acids are the substrates for the hexosamine biosynthetic pathway (HBP). This metabolic pathway generates the “sensing molecule” UDP-N-Acetylglucosamine (UDP-GlcNAc). UDP-GlcNAc is the substrate for the enzymes involved in protein N- and O-glycosylation, two important post-translational modifications (PTMs) identified in several proteins localized in the extracellular space, on the cell membrane and in the cytoplasm, nucleus and mitochondria. Since protein glycosylation controls several key aspects of cell physiology, aberrant protein glycosylation has been associated with different human diseases, including cancer. Here we review recent evidence indicating the tight association between the HBP flux and cell metabolism, with particular emphasis on the post-transcriptional and transcriptional mechanisms regulated by the HBP that may cause the metabolic rewiring observed in cancer. We describe the implications of both protein O- and N-glycosylation in cancer cell metabolism and bioenergetics; focusing our attention on the effect of these PTMs on nutrient transport and on the transcriptional regulation and function of cancer-specific metabolic pathways

Transcriptional profiling of immortalized and K-ras-transformed mouse fibroblasts upon PKA stimulation by forskolin in low glucose availability

Forskolin (FSK) induces activation of protein kinase A (PKA). This activation protects specifically some cancer cells from death induced by glucose starvation. Cell effects upon FSK treatment prompted us to investigate in detail the physiological role of PKA in the activation of pro-survival mechanisms in glucose starvation. In this regard we performed a microarray analysis of normal NIH3T3 and transformed NIH3T3-K-ras mouse fibroblasts cultured at 1 mM glucose and daily treated or not with 10 μM FSK until 72 h of growth, when the samples were collected. The microarray is deposited into Gene Expression Omnibus under Series GSE68266. The microarray data revealed that the activation of PKA regulates the expression of genes involved in metabolic, stress-response and pro-survival processes, like glutamine metabolism, autophagy and unfolded protein response, preventing cancer cell death in glucose starvation. Altogether these findings suggest that PKA activation, by inducing a complex transcriptional program, leads to cancer survival in nutrient stress, a typical feature of developing tumor. These transcriptional data, identifying this important role of PKA, will be useful to identify novel target in cancer therapy

New Insight and Knowledge on anti-inflammatory Effectiveness of dietary phenolics: the NIKE Project

The NIKE project is targeted to evaluate the impact of dietary phenolics\u2019 intake on the maintenance of remission in patients with inflammatory bowel disease (IBD). The project is particularly focused on a (poly)phenolic subclass, namely ellagitannins (ETs), mainly found in some fruits and nuts, like berries, pomegranate, and walnuts. Differently from common approach, the dietary intervention will be conducted at the beginning of the project. As first step, the in vivo metabolites and their concentration in the bloodstream - after consumption of the ET-rich food - will be assessed. A commercially available pomegranate juice will be used because its great richness in ETs. Volunteers suffering of asymptomatic IBD with high risk of clinical flare will be recruited at the St. Orsola-Malpighi Hospital (Bologna, Italy). The level of faecal calprotectin will be used as surrogate marker of the intestinal inflammation [1]. As second step, ET metabolites found in blood will be used for supplementation in two cell model systems (intestinal and immune cells), at concentration comparable to the in vivo ones, to unravel the mechanism/s of anti-inflammatory action. Signalling pathways potentially affected by ET metabolites in inflammatory conditions will be studied. In the third step, data obtained in cell cultures will be verified ex-vivo in tissue samples (intestinal biopsies and plasma) obtained during the intervention study. To our knowledge, NIKE will be the first project investigating the putative protective effects of these compounds toward inflammation and consequent IBD relapse in humans. At present only one study has investigated the anti-inflammatory effects of ETs related to IBD in rats [2]. The integral analysis of all results obtained in the project will elucidate the role of ET-rich foods in the secondary prevention of IBD, deepening the existing knowledge on their mechanism/s of action at the molecular, metabolic, and genomic levels. Acknowledgments. This study was funded by the Italian Ministry of Education, University and Research MIUR - SIR Programme no. RBSI14LHMB funded to F.D. References. 1) Konikoff & Denson, Inflamm Bowel Dis 2006; 12:524-34. 2) Rosillo et al, Pharmacol Res 2012; 66:235-42

Pomegranate: from the Promised Land to calm the fire in Inflammatory Bowel Disease

Inflammatory bowel disease (IBD) has a multifactorial aetiology and it is thought to be related to a combination of individual genetic susceptibility and environmental triggers that stimulate an inflammatory response. Although evidence indicates that dietary intake plays an important role, few studies have focused on the effect of (poly)phenol-rich food consumption on early markers of mucosal inflammation. At present only one study has investigated the anti-inflammatory effects of ellagitannins (ETs), a (poly)phenolic subclass mainly found in some fruits and nuts, related to IBD in rats [1]. Therefore, we hypothesised that treatment with a fruit juice containing ETs can significantly modulate biomarkers of mucosal inflammation compared with a control group receiving placebo. The double-blind, randomised controlled trial will include patients with IBD, ulcerative colitis and Crohn's disease, in stable clinical remission (≥3 months). Participants will be randomly allocated to one of two groups: active treatment (125 mL pomegranate juice, naturally rich in ETs) or placebo taken twice daily for 12 weeks. The primary outcome is to measure changes to the faecal neutrophil-derived protein calprotectin, surrogate marker of mucosal improvement, between the two groups from baseline to 12 weeks later. The secondary outcomes include systemic and mucosal changes of biochemical and molecular inflammatory response markers. The compliance of trial participants will be tested by uHPLC system coupled to mass spectrometer to quantify ET-metabolites in plasma and urine. In addition to the primary and secondary objectives, this trial will include plasma level of trimethylamine-N-oxide (TMAO) that may have potential as a biomarker to assess disease activity in IBD [2]. References. 1. Rosillo et al. 2012 Pharmacol Res. 2012; 66:235-42. 2. Wilson et al. 2015 Dig Dis Sci. 60:3620-30. Acknowledgments. This study was funded by the Italian Ministry of Education, University and Research MIUR - SIR Programme no. RBSI14LHMB funded to F.D. All beverages will be provided by Conserve Italia (Bologna, Italy)

Polifenoli ed efficacia antinfiammatoria: lo strano caso del melograno e le malattie croniche intestinali

Premesse. Il Progetto NIKE \u201cNew Insight and Knowledge on anti-inflammatory Effectiveness of dietary phenolics\u201d \ue8 un progetto finanziato Ministero dell\u2019Istruzione, dell\u2019Universit\ue0 e della Ricerca nell\u2019ambito del Programma SIR (Scientific Independence of young Researchers). Obiettivi. Focus generale del progetto \ue8 studiare l\u2019efficacia protettiva anti-infiammatoria di un alimento naturalmente ricco in ellagitannini (ET) - una sottoclasse di polifenoli - verso l\u2019infiammazione, al fine garantire una remissione clinica pi\uf9 prolungata nelle malattie infiammatorie croniche intestinali (inflammatory bowel disease, IBD), quali morbo di Crohn e rettocolite ulcerosa. Inoltre, NIKE si propone di chiarire il meccanismo di azione che sottende tali effetti anti-infiammatori a livello cellulare e dell\u2019intero organismo. Metodi. Uno studio randomizzato controllato con placebo sar\ue0 condotto all\u2019inizio del progetto presso l\u2019Ospedale Sant\u2019Orsola-Malpighi di Bologna al fine di valutare gli effetti della somministrazione di un succo di melagrana, naturalmente ricco in ET, su di un gruppo di soggetti con IBD in remissione ad alto rischio di recidiva. Il livello di calprotectina fecale sar\ue0 utilizzato come indice di attivit\ue0 dei processi flogistici intestinali. I metaboliti degli ET saranno identificati e misurati in plasma, urine e feci come marker dell\u2019assunzione dietetica. Successivamente, i metaboliti identificati nel sangue saranno utilizzati per la supplementazione in due tipi di colture cellulari (cellule intestinali e immunitarie), a concentrazioni paragonabili a quelle misurate in vivo, per indagarne il/i meccanismo/i di azione dell\u2019ipotizzato effetto anti-infiammatorio. I dati ottenuti in vitro saranno infine verificati ex-vivo in campioni di tessuti (biopsie intestinali e plasma) ottenuti durante lo studio di intervento. Risultati attesi. Fino ad oggi, un solo studio ha esaminato gli effetti anti-infiammatori degli ET relativi alle IBD nell\u2019animale da esperimento [1]. Quindi NIKE sar\ue0 il primo progetto a indagare gli effetti protettivi putativi di questi composti verso l\u2019infiammazione e conseguente ricaduta di IBD nell\u2019uomo. Inoltre, l\u2019analisi integrata di tutti i risultati ottenuti nell\u2019ambito del progetto consentir\ue0 ad aumentare le conoscenze su biodisponibilit\ue0, bioattivit\ue0 e meccanismo di azione degli ET. Questo studio \ue8 finanziato dal Ministero dell\u2019Istruzione, dell\u2019Universit\ue0 e della Ricerca (MIUR) nell\u2019ambito del Programma SIR 2014 (Progetto RBSI14LHMB, responsabile F.D.). [1] Rosillo et al, Pharmacol Res 2012; 66:235-42