Metformin Increases Natural Killer Cell Functions in Head and Neck Squamous Cell Carcinoma Through CXCL1 Inhibition

BACKGROUND: Metformin slows tumor growth and progression in vitro, and in combination with chemoradiotherapy, resulted in high overall survival in patients with head and neck cancer squamous cell carcinoma (HNSCC) in our phase 1 clinical trial (NCT02325401). Metformin is also postulated to activate an antitumor immune response. Here, we investigate immunologic effects of metformin on natural killer (NK) and natural killer T cells, including results from two phase I open-label studies in patients with HNSCC treated with metformin (NCT02325401, NCT02083692). METHODS: Peripheral blood was collected before and after metformin treatment or from newly diagnosed patients with HNSCC. Peripheral immune cell phenotypes were evaluated using flow cytometry, cytokine expression by ELISA and/or IsoLight, and NK cell-mediated cytotoxicity was determined with a flow-based NK cell cytotoxicity assay (NKCA). Patient tumor immune infiltration before and after metformin treatment was analyzed with immunofluorescence. NK cells were treated with either vehicle or metformin and analyzed by RNA sequencing (RNA-seq). NK cells were then treated with inhibitors of significant pathways determined by RNA-seq and analyzed by NKCA, ELISA, and western blot analyses. RESULTS: Increased peripheral NK cell activated populations were observed in patients treated with metformin. NK cell tumor infiltration was enhanced in patients with HNSCC treated with metformin preoperatively. Metformin increased antitumorigenic cytokines ex vivo, including significant increases in perforin. Metformin increased HNSCC NK cell cytotoxicity and inhibited the CXCL1 pathway while stimulating the STAT1 pathway within HNSCC NK cells. Exogenous CXCL1 prevented metformin-enhanced NK cell-mediated cytotoxicity. Metformin-mediated NK cell cytotoxicity was found to be AMP-activated protein kinase independent, but dependent on both mechanistic target of rapamycin and pSTAT1. CONCLUSIONS: Our data identifies a new role for metformin-mediated immune antitumorigenic function through NK cell-mediated cytotoxicity and downregulation of CXCL1 in HNSCC. These findings will inform future immunomodulating therapies in HNSCC

Psychological treatments and psychotherapies in the neurorehabilitation of pain. Evidences and recommendations from the italian consensus conference on pain in neurorehabilitation

BACKGROUND: It is increasingly recognized that treating pain is crucial for effective care within neurological rehabilitation in the setting of the neurological rehabilitation. The Italian Consensus Conference on Pain in Neurorehabilitation was constituted with the purpose identifying best practices for us in this context. Along with drug therapies and physical interventions, psychological treatments have been proven to be some of the most valuable tools that can be used within a multidisciplinary approach for fostering a reduction in pain intensity. However, there is a need to elucidate what forms of psychotherapy could be effectively matched with the specific pathologies that are typically addressed by neurorehabilitation teams. OBJECTIVES: To extensively assess the available evidence which supports the use of psychological therapies for pain reduction in neurological diseases. METHODS: A systematic review of the studies evaluating the effect of psychotherapies on pain intensity in neurological disorders was performed through an electronic search using PUBMED, EMBASE, and the Cochrane Database of Systematic Reviews. Based on the level of evidence of the included studies, recommendations were outlined separately for the different conditions. RESULTS: The literature search yielded 2352 results and the final database included 400 articles. The overall strength of the recommendations was medium/low. The different forms of psychological interventions, including Cognitive-Behavioral Therapy, cognitive or behavioral techniques, Mindfulness, hypnosis, Acceptance and Commitment Therapy (ACT), Brief Interpersonal Therapy, virtual reality interventions, various forms of biofeedback and mirror therapy were found to be effective for pain reduction in pathologies such as musculoskeletal pain, fibromyalgia, Complex Regional Pain Syndrome, Central Post-Stroke pain, Phantom Limb Pain, pain secondary to Spinal Cord Injury, multiple sclerosis and other debilitating syndromes, diabetic neuropathy, Medically Unexplained Symptoms, migraine and headache. CONCLUSIONS: Psychological interventions and psychotherapies are safe and effective treatments that can be used within an integrated approach for patients undergoing neurological rehabilitation for pain. The different interventions can be specifically selected depending on the disease being treated. A table of evidence and recommendations from the Italian Consensus Conference on Pain in Neurorehabilitation is also provided in the final part of the pape

Metropole video shop management system

A routinized function in a video shop at Makati was automated by developing an easy to use software written in DBASE III on an IBM PC XT. It assists in the management of a video shop along with custome

Metformin increases natural killer cell functions in head and neck squamous cell carcinoma through CXCL1 inhibition

BackgroundMetformin slows tumor growth and progression in vitro, and in combination with chemoradiotherapy, resulted in high overall survival in patients with head and neck cancer squamous cell carcinoma (HNSCC) in our phase 1 clinical trial (NCT02325401). Metformin is also postulated to activate an antitumor immune response. Here, we investigate immunologic effects of metformin on natural killer (NK) and natural killer T cells, including results from two phase I open-label studies in patients with HNSCC treated with metformin (NCT02325401, NCT02083692).MethodsPeripheral blood was collected before and after metformin treatment or from newly diagnosed patients with HNSCC. Peripheral immune cell phenotypes were evaluated using flow cytometry, cytokine expression by ELISA and/or IsoLight, and NK cell-mediated cytotoxicity was determined with a flow-based NK cell cytotoxicity assay (NKCA). Patient tumor immune infiltration before and after metformin treatment was analyzed with immunofluorescence. NK cells were treated with either vehicle or metformin and analyzed by RNA sequencing (RNA-seq). NK cells were then treated with inhibitors of significant pathways determined by RNA-seq and analyzed by NKCA, ELISA, and western blot analyses.ResultsIncreased peripheral NK cell activated populations were observed in patients treated with metformin. NK cell tumor infiltration was enhanced in patients with HNSCC treated with metformin preoperatively. Metformin increased antitumorigenic cytokines ex vivo, including significant increases in perforin. Metformin increased HNSCC NK cell cytotoxicity and inhibited the CXCL1 pathway while stimulating the STAT1 pathway within HNSCC NK cells. Exogenous CXCL1 prevented metformin-enhanced NK cell-mediated cytotoxicity. Metformin-mediated NK cell cytotoxicity was found to be AMP-activated protein kinase independent, but dependent on both mechanistic target of rapamycin and pSTAT1.ConclusionsOur data identifies a new role for metformin-mediated immune antitumorigenic function through NK cell-mediated cytotoxicity and downregulation of CXCL1 in HNSCC. These findings will inform future immunomodulating therapies in HNSCC

Roman Liver Tx Allocation: Results at 48 months of an innovative regional allocationmodel based on MELDNa and Donor-to-Recipient Match

Roman Liver Tx Allocation: Results at 48 months of an innovative regional allocation model based on MELDNa and Donor-to-Recipient Match Federica Ferracci1, Alfonso W. Avolio1, Francesca R. Ponziani1, Antonio Grieco1, Salvatore Agnes1, Stefano Ginanni Corradini2, Fabio Melandro2, Flaminia Ferri2, Massimo Rossi2, Tommaso M. Manzia3, Ilaria Lenci3, Matteo Manuelli3, Mario Angelico3, Giuseppe Tisone3, Giovanni Vennarecci4, Marco Colasanti4, Roberto L. Meniconi4, Ubaldo Visco Comandini4, Giuseppe M. Ettorre4, Raffaella Zaccaria5, Nicola Torlone5, Maurizio Valeri5; 1Transplant center - University Hospital “A. Gemelli”, Rome, Italy; 2Transplant center - University Hospital “Umberto I”, Rome, Italy; 3Transplant center - University Hospital “Tor Vergata”, Rome, Italy; 4Transplant center - San Camillo Hospital, Rome, Italy; 5Transplant center - Regione Lazio, Rome, Italy BACKGROUND Liver allocation policies are different between Countries and regions. An innovative allocation model based on MELDNa and donor age was developed in Italy in a donor-to-recipient match perspective. 244 Liver transplants (LT) performed (2013-2015) in a 4 Center regional Consortium were compared to 235 controls (2010- 2012). P&M In the 2013-2015 period, Standard Organs (SO, N=151, 61,8%) were allocated to patients (pts) on a common regional waiting list, based on MELDNa (5 classes: >30, N=32, 13.2%; 29-24, N=41, 16.8%; 23-20, N=19, 7.8%; 19-15, N=24, 9.8%; <15, N=35, 14.3%). Non-SO (donor age >65, N=76, 31.2%;other reasons, N=17, 6.9%) were allocated locally, according to priority defined in each Center. and to individual patient’s characteristics (mainly HCC pts with progressive disease). In the 2010-2012 period each Center used a MELD based allocation algorithm and HCC pts were equipoised to non-HCC pts using the MELD 22 rule. Univariate Kaplan Meier (KM) and multivariate Cox analyses (Cox) were used. RESULTS LT in 2013-2015 period were older, with higher MELDNa, and received organs by older donors. Regarding non-HCC patients: Recipient’s age: 52±11 vs 50±11; Donor’s age: 51±18 vs 52±18; MELD at LT: 23±7 vs 21±7, p=0.002; MELDNa at LT: 25±7 vs 22±7, p=0.003. Regarding HCC patients: Recipient’s age: 58±6 vs 57±8; Donor’s age: 56±17 vs 52±18; MELD at LT: 15±6 vs 14±6; MELDNa at LT: 16±7 vs 15±5. Waiting time was significantly lower for non-HCC group 2.0±4.4 vs 6.7±8.3 mo, p=0.0001, and unchanged for the HCC patients 5.7±5.2 vs 5.3±5. 48 mo patient survival was similar (KM: overall 76% continuos l vs 79% dotted l; non-HCC: 78% vs 80%; HCC: 73% vs 77%). MELDNa and recipient’s age resulted significant prognostic predictors at Cox. CONCLUSIONS Although overall survival was slightly reduced in the non-SO HCC subgroup, the innovative allocation model provides positive results about overall LT performed, using organs from older donors, and faster transplantation for pts with progressive liver disease

Lack of reduction in serum alpha-fetoprotein during treatment with direct antiviral agents predicts hepatocellular carcinoma development in a large cohort of patients with hepatitis C virus-related cirrhosis

Risk of hepatocellular carcinoma (HCC) in hepatitis C virus cirrhotic patients treated with direct-acting antiviral agents (DAA) is still debating. We investigated it in a large cohort. The cohort comprised 1045 cirrhotic patients who completed treatment with DAA, with a median follow-up of 17.3\ua0months after end of treatment (EOT), including 943 patients without history of HCC and 102 previously treated for HCC. The majority were men (59.9%), with compensated cirrhosis (88.8%), genotype 1b (44.7%). Univariate, multivariate analysis and Kaplan-Meier curves were performed to detect predictors of HCC in patients with and without reduction in alpha-fetoprotein (AFP) during treatment. SVR12 was 95.6%. HCC developed in 95 (9.9%), including 54 of 943 (5.7%) occurrent and 41 of 102 (39%) recurrent tumours. De novo were more often unifocal (P\ua0=\ua00.01) and curable (P\ua0=\ua00.03). AFP decreased from 16.1\ua0\ub1\ua036.2\ua0mg/dL (baseline) to 11.4\ua0\ub1\ua055\ua0mg/dL (EOT). At univariate analysis, predictors were a previous HCC, older age, higher model for end-stage liver disease, prolonged INR, lower platelets, baseline and EOT AFP, virological failure and no reduction in AFP during treatment. Kaplan-Meier curves showed lower incidence of HCC in patients showing any reduction in AFP (P\ua0=\ua00.001). Those with AFP <6\ua0ng/mL had the lowest risk (P\ua0=\ua00.0002). At logistic regression, platelets (P\ua0=\ua00.009, OR 0.99 CI: 0.99-1.00), previous HCC (P\ua0<\ua00.000\ua001, OR: 10.76, 95% CI: 5.89-19.34) and no reduction in AFP during treatment (P\ua0=\ua00.0005, OR: 2.98, CI: 1.60-5.54) were independent predictors of HCC. In conclusion, risk of HCC after DAA treatment remains substantial. It is higher among patients with previous HCC, low platelets and without reduction in AFP during treatment

The Italian compassionate use of sofosbuvir in HCV patients waitlisted for liver transplantation: A national real-life experience

This study aimed to assess the real-life clinical and virological outcomes of HCV waitlisted patients for liver transplantation (LT) who received sofosbuvir/ribavirin (SOF/R) within the Italian compassionate use program. METHODS: Clinical and virological data were collected in 224 patients with decompensated cirrhosis and/or hepatocellular carcinoma (HCC) receiving daily SOF/R until LT or up a maximum of 48 weeks. RESULTS: Of 100 transplanted patients, 51 were HCV-RNA negative for >4 weeks before LT (SVR12: 88%) and 49 negative for <4 weeks or still viraemic at transplant: 34 patients continued treatment after LT (bridging therapy) (SVR12: 88%), while 15 stopped treatment (SVR12: 53%). 98 patients completed SOF/R without LT (SVR12: 73%). In patients with advanced decompensated cirrhosis (basal MELD 6515 and/or C-P 65B8), a marked improvement of the scores occurred in about 50% of cases and almost 20% of decompensated patients without HCC reached a condition suitable for inactivation and delisting. CONCLUSIONS: These real-life data indicate that in waitlisted patients: (i) bridging antiviral therapy can be an option for patients still viraemic or negative <4 weeks at LT; and (ii) clinical improvement to a condition suitable for delisting can occur even in patients with advanced decompensated cirrhosis