Bile duct cancer (BDC) is a malignancy thought to be derived from cholangiocytes, the epithelial cells lining the biliary tree. BDC is a highly aggressive tumour whose incidence has been increasing worldwide over the past two decades, now accounting for 10–15% of all hepato-biliary malignancies. Advanced BDC has a devastating prognosis, with a median survival of less than 24 months. The mechanisms underlying cholangiocyte malignant transformation and BDC progression is still not completely understood. Genomic profiling can offer a clearer understanding of their carcinogenesis, classification and treatment strategy. We performed large-scale genome sequencing analyses on BTCs to identify novel key-genes driving BDC and drug resistance. The long-term goals of this program are to understand the pathogenesis of BDC and develop new therapies for its treatment. We analyzed 100 BTC (Age: 50% >65y; Sex: 40% M, 60% F; Type: 30% iCCA, 30% dCCA; TNM: 11% I, 40% II, 43% III, 7% IV) samples from Modena Cohort, 40 by whole-exome sequencing (WES), 80 by RNA sequencing (RNAseq), and a further 30 samples by SmallRNA sequencing. We identified somatic alterations, transcriptomic and epigenetic profiles of tumours and stromal area of each sample, and searched for driver genes in BTCs. By using a bio-informatic pipeline, we integrated somatic mutation patterns and epigenetic features defined at the spatial level to identify novel target genes in the tumour microenvironment. Functional studies in 2D and 3D culturing models were conducted to investigate candidate genes linked to BDC progression. A total of 3392 and 6315 DEGs (Differentially expressed genes) were respectively observed in BDC comparing tumour (T), normal (N) and stromal (ST) areas with the criterion of false discovery rate < 0.05. In top-ranked differentially regulated gene sets, we identified primary cilium-associated genes (PC). OFD1, CNGB1, AURKA, CENPF, STIL, STK39, RAB23 and OSR1 were found based on the criteria of fold change > 2.5 and P < 0.01. We started also to clarify at molecular level the role of PC in BDC pathogenesis and progression. A therapeutic approach targeting OFD1 in BDC cells was also investigated. We performed large-scale genome sequencing analysis of 100 BTC samples from Italian populations. Among top-ranked dis-regulated gene sets we identified ciliary-associated genes. Loss of PC is frequently observed in BDC, suggesting that the absence of this organelle may promote tumorigenesis through aberrant signal transduction and the inability to exit the cell cycle. We investigated the molecular mechanisms underlying the cilia loss and test whether may be potential therapeutic target. These findings could be useful to establish treatment and diagnostic strategies for BTCs based on genetic information. The proposed research is relevant to public health because understanding and manipulating asymmetric ciliary signaling in cancer cells and cells of the tumour microenvironment can potentially improve understanding of cancer pathogenesis, and application of anti-cancer therapies.

A complete tumor response to neoadjuvant chemo-radiation (CRT) for locally advanced rectal cancer (LARC) is associated with better outcomes, but it is achieved in only 20-30% of patients (pts). The potential role of concomitant medications in addition to CRT has been evaluated in many studies to enhance response rate, but data are not definitive. In this multicentric, retrospective study, we investigated the influence of various concomitant medications on outcomes in this setting of pts. 246 pts treated at Azienda Ospedaliero-Universitaria of Modena, University of Parma, and Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori from 2003 to 2018 were retrospectively identified. Demographical and clinicopathological data of potential interest were collected. Odds Ratio (OR) was used to assess the association between the use of concomitant drugs and outcomes (Dworak tumor regression grade [TRG] and disease-free survival [DFS]). All pts received neoadjuvant CRT. Of them, 15.8% were taking antihypertensive drugs, 3.7% statins, 1.2% antiplatelet drugs, and 1.6% antidiabetic drugs. Moreover, 30.5% of pts were taking ≥2 of these drugs simultaneously. Roughly 40% of pts have experienced CRT-related toxicity, leading to a dose reduction or treatment discontinuation in 27% of cases. Of the 221 surgical specimens available, a Dworak TRG score 3-4 was achieved in 30.7% of cases. At both univariate and multivariate analysis, we found an association between statins and a Dworak 3-4 (OR 8.78, p=0.01). Furthermore, statins were also significantly associated with more frequently CRT-related toxicity (OR 2.39, p=0.0098) and a more frequent chemotherapy dose reduction or discontinuation (OR 2.26, p=0.03). No correlations were found between the use of the evaluated drugs and the DFS. Despite a higher frequency of chemotherapy interruption or dose reduction and radiotherapy interruption, in this series of pts with LARC treated with neoadjuvant CRT, the concomitant use of statins proved to be associated with better tumor regression, according to the Dworak system. Larger prospective trials are awaited to confirm this hypothesis.

The efficacy of Ramucirumab in the treatment of advanced gastric cancer was demonstrated in the two pivotal study REGARD and RAINBOW, which both demonstrated significant improvements in Overall Survival (OS) and Progression-Free Survival (PFS). However, data on validated clinical and biochemical predictive factors in a real-life setting are still lacking. Retrospective analysis of medical records of patients with advanced gastric cancers treated with ramucirumab or ramucirumab/paclitaxel from April 2015 to April 2020 at Modena Cancer Center was conducted. Clinicopathological and biochemical parameters deemed of interest were collected. The cut-off value for continuous variables was assessed at 75° percentile. Treatment effects were evaluated by univariate and multivariate Cox proportional hazards regression models for OS and logistic regression analysis. During the study period, 49 patients with advanced gastric cancers were treated with ramucirumab alone (6.1%) or in combination with paclitaxel (93.9%) as second line therapy. At the multivariate, only CEA showed to impact on the second-line PFS with statistical significance (p=0.013, 95% Confidence Interval [CI] 1.08-6.4). By performing a logistic regression analysis, Absolute Neutrophil Count (ANC), Neutrophil Lymphocyte Ratio (NLR), Systemic Inflammatory Index (SII), CEA, Body Mass Index (BMI), ECOG Performance Status and Clinical Benefit (defined as complete response, partial response and stable disease versus disease progression) showed to be correlate with a PFS of more than 6 months. ECOG 0 and BMI>25 were the main predictive factors associated with Clinical Benefit (p=0.018 and p= 0.0006, respectively). Favorable inflammatory indexes, CEA, BMI and ECOG confer a better second-line PFS. ECOG and BMI seem to be strictly related to Clinical Benefit, thus making these two parameters predictive of response to Ramucirumab. In the future, it will be mandatory a validation of our results on a larger population by the use of more specific parameters for the sarcopenia assessment.

Background ABC portend a dismal prognosis despite standard chemotherapy treatment. A proper risk-stratification is key to optimize the benefit-to-risk ratio of palliative treatment. The PNI is an immune-inflammatory and nutritional indicator showing to be prognostic across a number of malignancies. We aimed at investigating the impact on survival of the PNI in ABC patients (pts) treated with 1L. Methods Electronic medical records of pts diagnosed with ABC and treated with 1L between 2002 and 2018 at the Modena Cancer Centre were retrospectively reviewed. Clinical, pathological and biochemical variables of potential interest were collected. The PNI was calculated as follows: 10 × serum albumin concentration (g/dL) + 0.005 × peripheral lymphocyte count (number/mm2) and dichotomized using the ROC analysis with 36.7 as the cut-off value. Univariate and multivariate analysis were performed to assess the impact of covariates on overall survival (OS). Kaplan-Meier survival curves were generated and log-rank testing was used to make comparisons. Results Overall, 114 pts fulfilled the inclusion criteria and were included in the analysis. 51% (n = 58) were female and 49% (n = 56) had an ECOG PS of 0. 35% (n = 40) of pts received a platinum/gemcitabine doublet, while 65% (n = 74) were treated with other regimens. The median OS in the cohort was 8.1 months. At the univariate analysis the following covariates were associated with OS: PNI (P 36.7 and Conclusions We demonstrated an independent prognostic role for the PNI in a cohort of ABC treated with 1L. Since it is based on easy-to-collect and inexpensive parameters it should be implemented in the clinical practice to improve the accuracy of current available tools. Legal entity responsible for the study Francesco Caputo. Funding Has not received any funding. Disclosure All authors have declared no conflicts of interest.

Background: Recent studies suggest that primary tumour location (PTL) has a prognostic value in patients (pts) with metastatic colorectal cancer (CRC), while its role in early stage disease remains unclear. The aim of this analysis is to investigate the relationship between PTL and the development of metachronous metastasis. Methods: We performed a population-based study from Modena Cancer Registry collecting data of patients (pts) with early stage disease (stage I, II, III) who underwent surgery from 1995 to 2010. We hypothesized a potential impact of PTL on the postoperative recurrence rate. Fisher's exact test, univariate and multivariate Cox regression analysis were performed. Results: During the study period, 1570 pts with left-sided colon cancer (LCC) and 841 pts with right-sided colon cancer (RCC) were registered. In the entire cohort, 268 of 1576 pts (17%) with LCC and 100 of 841 pts (11.2 %) with RCC developed metachronous metastasis, for a total of 368 of 2411 pts (15%). Comparing LCC and RCC clinical and pathological status we found no statistically difference in lymph-node status (p = 0.737) but an increasing rate of G3 cancers in RCC vs LCC (p = 0.010). Median overall survival (OS) from early stage disease diagnosis for LCC patients was 45 months versus 35 months for RCC patients, with no significant difference in relapse free survival between the two groups (23.8 Vs 23.0 months). When relapsed, time to death resulted to be significantly longer in LCC group than in RCC group (14.7 vs 6.3 months; HR 1.46, 95% C.I. 1,16-1,86; p 0,001). In the multivariate Cox regression analysis adjusted for grading and stage at diagnosis, we confirmed a statistically significant impact of the primary tumour sidedness on OS in the relapsed setting (HR 1.48, 95% C.I: 1.15-1.89, p = 0.001). Conclusions: In accordance to literature, our registry data confirm the prognostic role of PTL in advanced colorectal cancers: in particular, right-sided tumours have low recurrence rate but poor prognosis once relapsed. Other investigations are necessary to better understand the substantial heterogeneity within the molecular biology of RCC and LCC in order to provide a better post-operative surveillance and to select the most effective treatment strategies after relapse. Legal entity responsible for the study: Modena University Hospital. Funding: Has not received any funding. Disclosure: All authors have declared no conflicts of interest.