Non-mitogenic FGF19 mRNA-based therapy for the treatment of experimental metabolic dysfunction-associated steatotic liver disease (MASLD).

Metabolic dysfunction-associated steatohepatitis (MASH) represents a global health threat. MASH pathophysiology involves hepatic lipid accumulation and progression to severe conditions like cirrhosis and, eventually, hepatocellular carcinoma. Fibroblast growth factor (FGF)-19 has emerged as a key regulator of metabolism, offering potential therapeutic avenues for MASH and associated disorders. We evaluated the therapeutic potential of non-mitogenic (NM)-FGF19 mRNA formulated in liver-targeted lipid nanoparticles (NM-FGF19-mRNAs-LNPs) in C57BL/6NTac male mice with diet-induced obesity and MASH (DIO-MASH: 40% kcal fat, 20% kcal fructose, 2% cholesterol). After feeding this diet for 21 weeks, NM-FGF19-mRNAs-LNPs or control (C-mRNA-LNPs) were administered (0.5 mg/kg, i.v.) weekly for another six weeks, in which diet feeding continued. NM-FGF19-mRNAs-LNPs treatment in DIO-MASH mice resulted in reduced body weight, adipose tissue depots, and serum transaminases, along with improved insulin sensitivity. Histological analyses confirmed the reversal of MASH features, including steatosis reduction without worsening fibrosis. NM-FGF19-mRNAs-LNPs reduced total hepatic bile acids (BAs) and changed liver BA composition, markedly influencing cholesterol homeostasis and metabolic pathways as observed in transcriptomic analyses. Extrahepatic effects included the down-regulation of metabolic dysfunction-associated genes in adipose tissue. This study highlights the potential of NM-FGF19-mRNA-LNPs therapy for MASH, addressing both hepatic and systemic metabolic dysregulation. NM-FGF19-mRNA demonstrates efficacy in reducing liver steatosis, improving metabolic parameters, and modulating BA levels and composition. Given the central role played by BA in dietary fat absorption, this effect of NM-FGF19-mRNA may be mechanistically relevant. Our study underscores the high translational potential of mRNA-based therapies in addressing the multifaceted landscape of MASH and associated metabolic perturbations.

Identification of PRMT5 as a therapeutic target in cholangiocarcinoma.

BACKGROUND: Cholangiocarcinoma (CCA) is a very difficult-to-treat cancer. Chemotherapies are little effective and response to immune checkpoint inhibitors is limited. Therefore, new therapeutic strategies need to be identified. OBJECTIVE: We characterised the enzyme protein arginine-methyltransferase 5 (PRMT5) as a novel therapeutic target in CCA. DESIGN: We evaluated the expression of PRMT5, its functional partner MEP50 and methylthioadenosine phosphorylase (MTAP)-an enzyme that modulates the sensitivity of PRMT5 to pharmacological inhibitors-in human CCA tissues. PRMT5-targeting drugs, currently tested in clinical trials for other malignancies, were assessed in human CCA cell lines and organoids, as well as in two immunocompetent CCA mouse models. Transcriptomic, proteomic and functional analyses were performed to explore the underlying antitumoural mechanisms. RESULTS: PRMT5 and MEP50 proteins were correlatively overexpressed in most CCA tissues. MTAP was absent in 25% of intrahepatic CCA. PRMT5-targeting drugs markedly inhibited CCA cell proliferation, synergising with cisplatin and gemcitabine and hindered the growth of cholangiocarcinoma organoids. PRMT5 inhibition blunted the expression of oncogenic genes involved in chromatin remodelling and DNA repair, consistently inducing the formation of RNA loops and promoting DNA damage. Treatment with PRMT5-targeting drugs significantly restrained the growth of experimental CCA without adverse effects and concomitantly induced the recruitment of CD4 and CD8 T cells to shrinking tumourous lesions. CONCLUSION: PRMT5 and MEP50 are frequently upregulated in human CCA, and PRMT5-targeting drugs have significant antitumoural efficacy in clinically relevant CCA models. Our findings support the evaluation of PRMT5 inhibitors in clinical trials, including their combination with cytotoxic and immune therapies.

Comprehensive analysis of epigenetic and epitranscriptomic genes' expression in human NAFLD.

Non-alcoholic fatty liver disease (NAFLD) is a multifactorial condition with a complex etiology. Its incidence is increasing globally in parallel with the obesity epidemic, and it is now considered the most common liver disease in Western countries. The precise mechanisms underlying the development and progression of NAFLD are complex and still poorly understood. The dysregulation of epigenetic and epitranscriptomic mechanisms is increasingly recognized to play pathogenic roles in multiple conditions, including chronic liver diseases. Here, we have performed a comprehensive analysis of the expression of epigenetic and epitranscriptomic genes in a total of 903 liver tissue samples corresponding to patients with normal liver, obese patients, and patients with non-alcoholic fatty liver (NAFL) and non-alcoholic steatohepatitis (NASH), advancing stages in NAFLD progression. We integrated ten transcriptomic datasets in an unbiased manner, enabling their robust analysis and comparison. We describe the complete landscape of epigenetic and epitranscriptomic genes' expression along the course of the disease. We identify signatures of genes significantly dysregulated in association with disease progression, particularly with liver fibrosis development. Most of these epigenetic and epitranscriptomic effectors have not been previously described in human NAFLD, and their altered expression may have pathogenic implications. We also performed a comprehensive analysis of the expression of enzymes involved in the metabolism of the substrates and cofactors of epigenetic and epitranscriptomic effectors. This study provides novel information on NAFLD pathogenesis and may also guide the identification of drug targets to treat this condition and its progression towards hepatocellular carcinoma.

Identification and experimental validation of druggable epigenetic targets in hepatoblastoma.

BACKGROUND & AIMS: Hepatoblastoma (HB) is the most frequent childhood liver cancer. Patients with aggressive tumors have limited therapeutic options; therefore, a better understanding of HB pathogenesis is needed to improve treatment. HBs have a very low mutational burden; however, epigenetic alterations are increasingly recognized. We aimed to identify epigenetic regulators consistently dysregulated in HB and to evaluate the therapeutic efficacy of their targeting in clinically relevant models. METHODS: We performed a comprehensive transcriptomic analysis of 180 epigenetic genes. Data from fetal, pediatric, adult, peritumoral (n = 72) and tumoral (n = 91) tissues were integrated. Selected epigenetic drugs were tested in HB cells. The most relevant epigenetic target identified was validated in primary HB cells, HB organoids, a patient-derived xenograft model, and a genetic mouse model. Transcriptomic, proteomic and metabolomic mechanistic analyses were performed. RESULTS: Altered expression of genes regulating DNA methylation and histone modifications was consistently observed in association with molecular and clinical features of poor prognosis. The histone methyltransferase G9a was markedly upregulated in tumors with epigenetic and transcriptomic traits of increased malignancy. Pharmacological targeting of G9a significantly inhibited growth of HB cells, organoids and patient-derived xenografts. Development of HB induced by oncogenic forms of ß-catenin and YAP1 was ablated in mice with hepatocyte-specific deletion of G9a. We observed that HBs undergo significant transcriptional rewiring in genes involved in amino acid metabolism and ribosomal biogenesis. G9a inhibition counteracted these pro-tumorigenic adaptations. Mechanistically, G9a targeting potently repressed the expression of c-MYC and ATF4, master regulators of HB metabolic reprogramming. CONCLUSIONS: HBs display a profound dysregulation of the epigenetic machinery. Pharmacological targeting of key epigenetic effectors exposes metabolic vulnerabilities that can be leveraged to improve the treatment of these patients. IMPACT AND IMPLICATIONS: In spite of recent advances in the management of hepatoblastoma (HB), treatment resistance and drug toxicity are still major concerns. This systematic study reveals the remarkable dysregulation in the expression of epigenetic genes in HB tissues. Through pharmacological and genetic experimental approaches, we demonstrate that the histone-lysine-methyltransferase G9a is an excellent drug target in HB, which can also be harnessed to enhance the efficacy of chemotherapy. Furthermore, our study highlights the profound pro-tumorigenic metabolic rewiring of HB cells orchestrated by G9a in coordination with the c-MYC oncogene. From a broader perspective, our findings suggest that anti-G9a therapies may also be effective in other c-MYC-dependent tumors.

Mapping early serum proteome signatures of liver regeneration in living donor liver transplant cases.

The liver is the only solid organ capable of regenerating itself to regain 100% of its mass and function after liver injury and/or partial hepatectomy (PH). This exceptional property represents a therapeutic opportunity for severe liver disease patients. However, liver regeneration (LR) might fail due to poorly understood causes. Here, we have investigated the regulation of liver proteome and phosphoproteome at a short time after PH (9 h), to depict a detailed mechanistic background of the early LR phase. Furthermore, we analyzed the dynamic changes of the serum proteome and metabolome of healthy living donor liver transplant (LDLT) donors at different time points after surgery. The molecular profiles from both analyses were then correlated. Insulin and FXR-FGF15/19 signaling were stimulated in mouse liver after PH, leading to the activation of the main intermediary kinases (AKT and ERK). Besides, inhibition of the hippo pathway led to an increased expression of its target genes and of one of its intermediary proteins (14-3-3 protein), contributing to cell proliferation. In association with these processes, metabolic reprogramming coupled to enhanced mitochondrial activity cope for the energy and biosynthetic requirements of LR. In human serum of LDLT donors, we identified 56 proteins and 13 metabolites statistically differential which recapitulate some of the main cellular processes orchestrating LR in its early phase. These results provide mechanisms and protein mediators of LR that might prove useful for the follow-up of the regenerative process in the liver after PH as well as preventing the occurrence of complications associated with liver resection.

New molecular mechanisms in cholangiocarcinoma: signals triggering interleukin-6 production in tumor cells and KRAS co-opted epigenetic mediators driving metabolic reprogramming.

BACKGROUND: Cholangiocarcinoma (CCA) is still a deadly tumour. Histological and molecular aspects of thioacetamide (TAA)-induced intrahepatic CCA (iCCA) in rats mimic those of human iCCA. Carcinogenic changes and therapeutic vulnerabilities in CCA may be captured by molecular investigations in bile, where we performed bile proteomic and metabolomic analyses that help discovery yet unknown pathways relevant to human iCCA. METHODS: Cholangiocarcinogenesis was induced in rats (TAA) and mice (JnkΔhepa + CCl4 + DEN model). We performed proteomic and metabolomic analyses in bile from control and CCA-bearing rats. Differential expression was validated in rat and human CCAs. Mechanisms were addressed in human CCA cells, including Huh28-KRASG12D cells. Cell signaling, growth, gene regulation and [U-13C]-D-glucose-serine fluxomics analyses were performed. In vivo studies were performed in the clinically-relevant iCCA mouse model. RESULTS: Pathways related to inflammation, oxidative stress and glucose metabolism were identified by proteomic analysis. Oxidative stress and high amounts of the oncogenesis-supporting amino acids serine and glycine were discovered by metabolomic studies. Most relevant hits were confirmed in rat and human CCAs (TCGA). Activation of interleukin-6 (IL6) and epidermal growth factor receptor (EGFR) pathways, and key genes in cancer-related glucose metabolic reprogramming, were validated in TAA-CCAs. In TAA-CCAs, G9a, an epigenetic pro-tumorigenic writer, was also increased. We show that EGFR signaling and mutant KRASG12D can both activate IL6 production in CCA cells. Furthermore, phosphoglycerate dehydrogenase (PHGDH), the rate-limiting enzyme in serine-glycine pathway, was upregulated in human iCCA correlating with G9a expression. In a G9a activity-dependent manner, KRASG12D promoted PHGDH expression, glucose flow towards serine synthesis, and increased CCA cell viability. KRASG12D CAA cells were more sensitive to PHGDH and G9a inhibition than controls. In mouse iCCA, G9a pharmacological targeting reduced PHGDH expression. CONCLUSIONS: In CCA, we identified new pro-tumorigenic mechanisms: Activation of EGFR signaling or KRAS mutation drives IL6 expression in tumour cells; Glucose metabolism reprogramming in iCCA includes activation of the serine-glycine pathway; Mutant KRAS drives PHGDH expression in a G9a-dependent manner; PHGDH and G9a emerge as therapeutic targets in iCCA.

Gene supplementation of CYP27A1 in the liver restores bile acid metabolism in a mouse model of cerebrotendinous xanthomatosis.

Cerebrotendinous xanthomatosis (CTX) is an autosomal recessive disease caused by mutations in the CYP27A1 gene, encoding the sterol 27-hydroxylase. Disruption of the bile acid biosynthesis pathway and accumulation of toxic precursors such as cholestanol cause chronic diarrhea, bilateral juvenile cataracts, tissue deposition of cholestanol and cholesterol (xanthomas), and progressive motor/neuropsychiatric alterations. We have evaluated the therapeutic potential of adeno-associated virus (AAV) vectors expressing CYP27A1 in a CTX mouse model. We found that a vector equipped with a strong liver-specific promoter (albumin enhancer fused with the α1 anti-trypsin promoter) is well tolerated and shows therapeutic effect at relatively low doses (1.5 × 1012 viral genomes [vg]/kg), when less than 20% of hepatocytes overexpress the transgene. This vector restored bile acid metabolism and normalized the concentration of most bile acids in plasma. By contrast, standard treatment (oral chenodeoxycholic acid [CDCA]), while reducing cholestanol, did not normalize bile acid composition in plasma and resulted in supra-physiological levels of CDCA and its derivatives. At the transcriptional level, only the vector was able to avoid the induction of xenobiotic-induced pathways in mouse liver. In conclusion, the overexpression of CYP27A1 in a fraction of hepatocytes using AAV vectors is well tolerated and provides full metabolic restoration in Cyp27a1 -/- mice. These features make gene therapy a feasible option for the etiological treatment of CTX patients.

Splicing Factor SLU7 Prevents Oxidative Stress-Mediated Hepatocyte Nuclear Factor 4α Degradation, Preserving Hepatic Differentiation and Protecting From Liver Damage.

BACKGROUND AND AIMS: Hepatocellular dedifferentiation is emerging as an important determinant in liver disease progression. Preservation of mature hepatocyte identity relies on a set of key genes, predominantly the transcription factor hepatocyte nuclear factor 4α (HNF4α) but also splicing factors like SLU7. How these factors interact and become dysregulated and the impact of their impairment in driving liver disease are not fully understood. APPROACH AND RESULTS: Expression of SLU7 and that of the adult and oncofetal isoforms of HNF4α, driven by its promoter 1 (P1) and P2, respectively, was studied in diseased human and mouse livers. Hepatic function and damage response were analyzed in wild-type and Slu7-haploinsufficient/heterozygous (Slu7+/- ) mice undergoing chronic (CCl4 ) and acute (acetaminophen) injury. SLU7 expression was restored in CCl4 -injured mice using SLU7-expressing adeno-associated viruses (AAV-SLU7). The hepatocellular SLU7 interactome was characterized by mass spectrometry. Reduced SLU7 expression in human and mouse diseased livers correlated with a switch in HNF4α P1 to P2 usage. This response was reproduced in Slu7+/- mice, which displayed increased sensitivity to chronic and acute liver injury, enhanced oxidative stress, and marked impairment of hepatic functions. AAV-SLU7 infection prevented liver injury and hepatocellular dedifferentiation. Mechanistically we demonstrate a unique role for SLU7 in the preservation of HNF4α1 protein stability through its capacity to protect the liver against oxidative stress. SLU7 is herein identified as a key component of the stress granule proteome, an essential part of the cell's antioxidant machinery. CONCLUSIONS: Our results place SLU7 at the highest level of hepatocellular identity control, identifying SLU7 as a link between stress-protective mechanisms and liver differentiation. These findings emphasize the importance of the preservation of hepatic functions in the protection from liver injury.

ARMCX3 Mediates Susceptibility to Hepatic Tumorigenesis Promoted by Dietary Lipotoxicity.

ARMCX3 is encoded by a member of the Armcx gene family and is known to be involved in nervous system development and function. We found that ARMCX3 is markedly upregulated in mouse liver in response to high lipid availability, and that hepatic ARMCX3 is upregulated in patients with NAFLD and hepatocellular carcinoma (HCC). Mice were subjected to ARMCX3 invalidation (inducible ARMCX3 knockout) and then exposed to a high-fat diet and diethylnitrosamine-induced hepatocarcinogenesis. The effects of experimental ARMCX3 knockdown or overexpression in HCC cell lines were also analyzed. ARMCX3 invalidation protected mice against high-fat-diet-induced NAFLD and chemically induced hepatocarcinogenesis. ARMCX3 invalidation promoted apoptotic cell death and macrophage infiltration in livers of diethylnitrosamine-treated mice maintained on a high-fat diet. ARMCX3 downregulation reduced the viability, clonality and migration of HCC cell lines, whereas ARMCX3 overexpression caused the reciprocal effects. SOX9 was found to mediate the effects of ARMCX3 in hepatic cells, with the SOX9 interaction required for the effects of ARMCX3 on hepatic cell proliferation. In conclusion, ARMCX3 is identified as a novel molecular actor in liver physiopathology and carcinogenesis. ARMCX3 downregulation appears to protect against hepatocarcinogenesis, especially under conditions of high dietary lipid-mediated hepatic insult.

High mortality rate following periprosthetic femoral fractures after total hip arthroplasty. A multicenter retrospective study.

BACKGROUND: The main objective of this study was to evaluate the morbidity and mortality following periprosthetic femoral fractures (PFFs) after total hip arthroplasty. The secondary objectives were to explore risk factors for mortality and compare outcomes by method of treatment. METHODS: A multicenter retrospective study was conducted (2016-2017) of all PFFs after total hip arthroplasty. We collected data on: ASA score, Charlson comorbidity index, type of fracture, method of treatment, timing of surgery, length of stay, systemic and local complications and mortality. Functional outcome was assessed in terms of preoperative and postoperative ambulatory status. Univariate and multivariate analysis were performed in the sample to identify risk factors for mortality. RESULTS: A total of 107 patients were evaluated and their mean age was 81 years old. The most common type of fracture according to the Vancouver classification was B1 (52.4% of patients), followed by B2 fractures (31.8%). The mortality rate during the first month was 9.3% and was associated with patients with ASA >3. Mortality rate in the first year was 22.3% and was associated with poorer walking ability before surgery and Charlson index ≥3. In the multivariable analysis, Charlson index ≥3 (odds ratio = 6.85) and age ≥80 years old (odds ratio=7.446) were associated with 1-year mortality. Neither complications nor mortality rate were associated with either time to surgery or method of treatment. More than half of the patients (57.9%) did not regain their prefracture walking status. Major systemic complications developed in 23.4% of the patients and major local complications in 12.1%. CONCLUSION: Despite modern surgical techniques and multidisciplinary management, this study highlights the ambulatory status impairment and high rate of complications and mortality after PFF. Although the mortality rate during the first year was similar to that observed in other studies on PFFs, we found a higher mortality rate within the first month.

Dual-mobility cups in total hip arthroplasty after femoral neck fractures: A retrospective study comparing outcomes between cemented and cementless fixation.

INTRODUCTION: Total hip arthroplasty (THA) after femoral neck fracture (FNF) is associated with an increased risk of dislocation. The goals of our study were (1) to determine dislocation and revision rates when dual-mobility cups (DMCs) are used in these patients, (2) to analyze clinical and radiographic outcomes, survivorship, complications and mortality rate, and (3) to compare results between cemented and cementless cups. PATIENTS AND METHODS: We retrospectively reviewed patients with FNF treated using DMC-THA between 2011 and 2018. A minimum 2-year follow-up was required for clinical and radiographic assessment. The clinical outcome was assessed using the Harris Hip Score (HHS) and Merlé D´Aubigné-Postel score (MDP). Radiolucent lines, osteolysis and cup loosening were analyzed. RESULTS: We included 105 patients (105 hips) with a mean age of 75.5 years. There were no dislocations. One patient (1.0%) underwent cup revision at 39 months for aseptic cup loosening. The mean HHS and MDP were 80.5 and 14.2 respectively at a mean follow-up of 4.1 years. A higher MDP was found in patients with cementless rather than cemented cups (15.0 vs. 13.1; p = 0.006). Four patients had radiolucent lines > 1 mm, around cemented cups. At 6.8 years, estimated cup survival was 98.2% for revision for aseptic loosening and 97.3% for revision for any reason. The mortality rates were 6.7% at 1 year and 23.8% at last follow-up. CONCLUSION: Our findings suggest that using DMC in THA for FNF may prevent dislocation with a low revision rate. Cementless cups had a higher MDP than cemented cups.

Cementation of a dual-mobility cup into a well-fixed cementless shell in patients with high risk of dislocation undergoing revision total hip arthroplasty.

BACKGROUND: Cementation of polyethylene liners into well-fixed cementless metal shells has become an option during revision total hip arthroplasty (THA). We report the results of cementing a dual-mobility (DM) component into a stable acetabular shell in high-risk patients undergoing revision THA. METHODS: A single-centre series of 10 patients undergoing revision THA with a DM cup cemented into an existing well-fixed shell between 2012 and 2016 were retrospectively reviewed. Failure due to aseptic loosening or instability and implant survival at last follow-up were analysed. The average age was 79.2 years and mean follow-up was 3.5 years. Indications were recurrent hip dislocation in 8 cases and intraoperative instability with moderate abductor insufficiency in 2 cases. In cases with recurrent dislocation, the aetiology of instability was classified by Wera type. RESULTS: At the latest follow-up, Harris Hip Scores had improved from 49.3 preoperatively to 71.3 postoperatively (p = 0.098). In the 8 patients with recurrent dislocation, 4 cases (50%) had an unclear aetiology (Wera type 6), 2 (25%) abductor deficiency (Wera type 3) and 2 (25%) late polyethylene wear (type 5).Postoperative recurrent dislocation occurred in 1 hip (10%). No cases of intraprosthetic dislocation, aseptic loosening of the previous shell or dissociation at the cement-cup interface were identified. CONCLUSION: Although the follow-up of this series is short, cementation of a DM cup into a previous well-fixed socket seems to be a viable option to treat and prevent instability after revision THA, without providing constraint at the cement-cup interface.

Dual Targeting of G9a and DNA Methyltransferase-1 for the Treatment of Experimental Cholangiocarcinoma.

BACKGROUND AND AIMS: Cholangiocarcinoma (CCA) is a devastating disease often detected at advanced stages when surgery cannot be performed. Conventional and targeted systemic therapies perform poorly, and therefore effective drugs are urgently needed. Different epigenetic modifications occur in CCA and contribute to malignancy. Targeting epigenetic mechanisms may thus open therapeutic opportunities. However, modifications such as DNA and histone methylation often coexist and cooperate in carcinogenesis. We tested the therapeutic efficacy and mechanism of action of a class of dual G9a histone-methyltransferase and DNA-methyltransferase 1 (DNMT1) inhibitors. APPROACH AND RESULTS: Expression of G9a, DNMT1, and their molecular adaptor, ubiquitin-like with PHD and RING finger domains-1 (UHRF1), was determined in human CCA. We evaluated the effect of individual and combined pharmacological inhibition of G9a and DNMT1 on CCA cell growth. Our lead G9a/DNMT1 inhibitor, CM272, was tested in human CCA cells, patient-derived tumoroids and xenograft, and a mouse model of cholangiocarcinogenesis with hepatocellular deletion of c-Jun-N-terminal-kinase (Jnk)-1/2 and diethyl-nitrosamine (DEN) plus CCl4 treatment (JnkΔhepa + DEN + CCl4 mice). We found an increased and correlative expression of G9a, DNMT1, and UHRF1 in CCAs. Cotreatment with independent pharmacological inhibitors G9a and DNMT1 synergistically inhibited CCA cell growth. CM272 markedly reduced CCA cell proliferation and synergized with Cisplatin and the ERBB-targeted inhibitor, Lapatinib. CM272 inhibited CCA tumoroids and xenograft growth and significantly antagonized CCA progression in JnkΔhepa + DEN + CCl4 mice without apparent toxicity. Mechanistically, CM272 reprogrammed the tumoral metabolic transcriptome and phenotype toward a differentiated and quiescent status. CONCLUSIONS: Dual targeting of G9a and DNMT1 with epigenetic small molecule inhibitors such as CM272 is a potential strategy to treat CCA and/or enhance the efficacy of other systemic therapies.

FGF15/19 is required for adipose tissue plasticity in response to thermogenic adaptations.

OBJECTIVE: To determine the role of enterokine FGF15/19 in adipose tissue thermogenic adaptations. METHODS: Circulating FGF19 and gene expression (qRT-PCR) levels were assessed in subcutaneous adipose tissue from obese human patients. Effects of experimentally increased FGF15 and FGF19 levels in vivo were determined in mice using adenoviral and adeno-associated vectors. Adipose tissues were characterized in FGF15-null mice under distinct cold-related thermogenic challenges. The analyses spanned metabolic profiling, tissue characterization, histology, gene expression, and immunoblot assays. RESULTS: In humans, FGF19 levels are directly associated with UCP1 gene expression in subcutaneous adipose tissue. Experimental increases in FGF15 or FGF19 induced white fat browning in mice as demonstrated by the appearance of multilocular beige cells and markers indicative of a beige phenotype, including increased UCP1 protein levels. Mice lacking FGF15 showed markedly impaired white adipose tissue browning and a mild reduction in parameters indicative of BAT activity in response to cold-induced environmental thermogenic challenges. This was concomitant with signs of altered systemic metabolism, such as reduced glucose tolerance and impaired cold-induced insulin sensitization. CONCLUSIONS: Enterokine FGF15/19 is a key factor required for adipose tissue plasticity in response to thermogenic adaptations.

Dual Pharmacological Targeting of HDACs and PDE5 Inhibits Liver Disease Progression in a Mouse Model of Biliary Inflammation and Fibrosis.

Liver fibrosis, a common hallmark of chronic liver disease (CLD), is characterized by the accumulation of extracellular matrix secreted by activated hepatic fibroblasts and stellate cells (HSC). Fibrogenesis involves multiple cellular and molecular processes and is intimately linked to chronic hepatic inflammation. Importantly, it has been shown to promote the loss of liver function and liver carcinogenesis. No effective therapies for liver fibrosis are currently available. We examined the anti-fibrogenic potential of a new drug (CM414) that simultaneously inhibits histone deacetylases (HDACs), more precisely HDAC1, 2, and 3 (Class I) and HDAC6 (Class II) and stimulates the cyclic guanosine monophosphate (cGMP)-protein kinase G (PKG) pathway activity through phosphodiesterase 5 (PDE5) inhibition, two mechanisms independently involved in liver fibrosis. To this end, we treated Mdr2-KO mice, a clinically relevant model of liver inflammation and fibrosis, with our dual HDAC/PDE5 inhibitor CM414. We observed a decrease in the expression of fibrogenic markers and collagen deposition, together with a marked reduction in inflammation. No signs of hepatic or systemic toxicity were recorded. Mechanistic studies in cultured human HSC and cholangiocytes (LX2 and H69 cell lines, respectively) demonstrated that CM414 inhibited pro-fibrogenic and inflammatory responses, including those triggered by transforming growth factor ß (TGFß). Our study supports the notion that simultaneous targeting of pro-inflammatory and fibrogenic mechanisms controlled by HDACs and PDE5 with a single molecule, such as CM414, can be a new disease-modifying strategy.

Pilot Multi-Omic Analysis of Human Bile from Benign and Malignant Biliary Strictures: A Machine-Learning Approach.

Cholangiocarcinoma (CCA) and pancreatic adenocarcinoma (PDAC) may lead to the development of extrahepatic obstructive cholestasis. However, biliary stenoses can also be caused by benign conditions, and the identification of their etiology still remains a clinical challenge. We performed metabolomic and proteomic analyses of bile from patients with benign (n = 36) and malignant conditions, CCA (n = 36) or PDAC (n = 57), undergoing endoscopic retrograde cholangiopancreatography with the aim of characterizing bile composition in biliopancreatic disease and identifying biomarkers for the differential diagnosis of biliary strictures. Comprehensive analyses of lipids, bile acids and small molecules were carried out using mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (1H-NMR) in all patients. MS analysis of bile proteome was performed in five patients per group. We implemented artificial intelligence tools for the selection of biomarkers and algorithms with predictive capacity. Our machine-learning pipeline included the generation of synthetic data with properties of real data, the selection of potential biomarkers (metabolites or proteins) and their analysis with neural networks (NN). Selected biomarkers were then validated with real data. We identified panels of lipids (n = 10) and proteins (n = 5) that when analyzed with NN algorithms discriminated between patients with and without cancer with an unprecedented accuracy.

Revision hip arthroplasty with a rectangular tapered cementless stem: a retrospective study of the SLR-Plus stem at a mean follow-up of 4.1 years.

PURPOSE: Demand for revision total hip arthroplasty (THA) is growing, and this type of surgery remains challenging for orthopedic surgeons. Our objectives were to assess clinical and radiographic outcomes, survivorship and complications with the SLR-Plus stem in revision THA. METHODS: We retrospectively reviewed 65 patients (66 hips) who had undergone revision THA with the SLR-Plus stem between 2008 and 2015 at two medical institutions with a minimum 2-year follow-up. The clinical outcome was assessed using the Harris hip score and the Merlé D'Aubigné score preoperatively and at final follow-up. A postoperative visual analogue scale for pain and satisfaction was also used. Radiographic subsidence and fixation, Kaplan-Meier survivorship and complications were analyzed. RESULTS: The mean follow-up was 4.1 years (SD 2.1). Aseptic loosening (57.6%) was the main indication for surgery. The mean Harris Hip Score improved from 50.4 (SD 16.5) to 83 (SD 12.7) (p < 0.001) and mean Merlé D'Aubigné score improved from 9.5 (SD 2.7) to 14.3 (SD 2.2) (p < 0.001). A total of 98.4% of stems showed radiographically stable fixation. No aseptic loosening of the stem was seen. Radiolucent lines > 1 mm were observed in 33.3% of stems. Three stems were re-revised: two due to infection and one due to instability. At 7 years, estimated stem survival was 95.5% for revision for any reason and 100% for revision for aseptic loosening. Dislocation occurred in 7.6% of hips. CONCLUSION: We have shown significant clinical improvement, 98.4% of stable fixation and 100% stem survivorship for aseptic loosening in revision THA with the SLR-Plus stem.

Debridement, Antibiotics and Implant Retention for Early Periprosthetic Infections of the Hip: Outcomes and Influencing Factors.

PURPOSE: Debridement, antibiotics and implant retention (DAIR) is commonly performed and widely accepted for the treatment of acute infections following hip arthroplasty. The aims of this study were to: i) determine the DAIR success rate in treating acute postoperative and hematogenous periprosthetic infections of the hip at a tertiary hospital, ii) identify possible outcome predictors, and iii) analyze clinical and radiological outcomes. MATERIALS AND METHODS: We retrospectively reviewed cases of acute postoperative (≤3 months from index procedure) and hematogenous periprosthetic infections following total hip arthroplasty treated with DAIR at our hospital between 2004 and 2015. Overall, 26 hips (25 patients) were included in the study, with a mean age of 72.5 years (standard deviation [SD], 9.4). The mean follow-up was 48.5 months (SD, 43.7). Several variables (e.g., patient characteristics, infection type, surgery parameters) were examined to evaluate their influence on outcomes; functional and radiographic outcomes were assessed. RESULTS: The overall success rate of DAIR was 26.9%. The male sex was associated with treatment failure (P=0.005) and debridement performed by a surgeon in hip unit with success (P=0.028). DAIR failure increased in patients with chronic pulmonary disease (P=0.059) and steroid therapy (P=0.062). Symptom duration of <11 days until DAIR yielded a better infection eradication rate (P=0.068). The mean postoperative Harris Hip Score was 74.2 (SD, 16.6). CONCLUSION: DAIR, despite being used frequently, had a high failure rate in our series. Outcomes improved if an experienced hip arthroplasty surgeon performed the surgery. Patient comorbidities and symptom duration should be considered for decision-making.

DAIR (Debridement, Antibiotics and Implant Retention) less effective in hematogenous total knee arthroplasty infections.

BACKGROUND: Debridement and irrigation with prosthetic retention followed by antibiotic therapy (DAIR) is one of the treatments of choice in acute infections after a total knee arthroplasty. However, the success rate varies widely in the literature, depending on several factors such as comorbidities of the patient, duration of infection, and microorganisms involved. The goal of this study was to assess the outcomes of this therapeutic option and to identify possible predictors of the result. METHODS: We retrospectively reviewed cases of acute postoperative (≤ 3 months from index procedure) and acute hematogenous periprosthetic knee infections treated with DAIR at our hospital between 2004 and 2016. Overall, 26 knees were included, with a mean age of 73.4 years. Several variables related to patient characteristics, infection type, and surgery were examined to evaluate their influence on outcome, and functional and radiographic outcome were assessed. The mean follow-up was 41 months. A descriptive analysis was carried out on the collected data, and a univariate analysis was performed with the objective of searching for influential factors in the resolution of the infection using the chi-square nonparametric test in the case of the categorical variables and the Wilcoxon test for the continuous ones. Moreover, univariate cox regression analysis was performed. RESULTS: The overall success rate was 77% at the last follow-up, recording a significantly greater cure in acute infections (93% acute vs 58% acute hematogenous, p = 0.03). The infections in which the Staphylococcus aureus was isolated had a significantly lower cure rate, with only 33% of success, compared to 82% of the non-aureus microorganisms (p <  0.05). CONCLUSIONS: The present study shows a considerable cure rate in the treatment of acute knee infections through DAIR, although patient comorbidities, type of infection, and causative microorganism should be considered for decision-making.

A 65-Year Follow-up of a Judet Prosthesis.

The Judet polymethylmethacrylate hip hemiarthroplasty can be considered a major step in the development of the current total hip replacement. It was designed by the Judet brothers and first implanted in 1946. Although initial results were encouraging, the procedure was abandoned years later because of breakages and wear of the acrylic material. The authors present the case of an 85-year-old man in whom a Judet prosthesis was implanted 65 years ago after a traumatic injury of the left hip. He continued to work for 35 years after the arthroplasty and remained free of pain for an additional 5 years. The pain gradually intensified along with shortening of the left leg. The stem was removed in 2010 because it was protruding from the lateral aspect of the thigh, but no revision of the arthroplasty was performed because of the complexity of the surgery and the patient's age. At 65 years after the arthroplasty, the patient walked with a severe limp aided by 2 crutches and had a Harris Hip score of 30 points. Despite the eventual poor clinical outcome and the marked synovitis and bone loss present in the left hip, the authors believe this case deserves to be highlighted because of its longevity. This is the longest follow-up of a Judet prosthesis in the literature. To the authors' knowledge, just one other case of hip arthroplasty with such long follow-up has been reported. [Orthopedics. 2019; 42(3):e336-e338.].