High-grade Fibrosis Research Articles

Comparing imaging modalities in the assessment of fibrosis in metabolic dysfunction-associated steatotic liver disease.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most common liver disease in children. Liver biopsy is considered the gold standard for diagnosis. Magnetic resonance elastography (MRE) and transient elastography (TE) are imaging modalities that can monitor fibrosis and steatosis noninvasively. More studies are needed to identify whether imaging modalities can provide accurate and reproducible data. We hypothesize that MRE provides reliable data similar to that of TE when compared to liver biopsy in children with MASLD/metabolic dysfunction-associated steatohepatitis. We conducted a retrospective chart review of children with liver biopsy-proven MASLD at Children's Hospital Los Angeles between September 2017 and January 2023, investigating and comparing the predictive accuracy of MRE and TE in the detection of high-grade fibrosis on liver biopsy. Seventy-seven patients were reviewed, all of whom had undergone liver biopsy, MRE and TE for evaluation of MASLD. Fibrosis was identified in 90% of liver biopsies. The area under the receiver operating characteristic curves (AUROC) of MRE and TE for detection of high-grade fibrosis were 0.817 and 0.750, respectively, and not significantly different (p = 0.4785). We demonstrate that MRE and TE did not accurately predict high-grade fibrosis on liver biopsy. Between the two noninvasive imaging modalities, the correlation of identifying high-grade fibrosis was not statistically different; however, the AUROC for MRE was slightly superior to that of TE. Studies with larger cohorts will be required to validate these findings.

A long-acting LEAP2 analog reduces hepatic steatosis and inflammation and causes marked weight loss in mice

ObjectiveThe number of individuals affected by metabolic dysfunction associated fatty liver disease [1] is on the rise, yet hormonal contributors to the condition remain incompletely described and only a single FDA-approved treatment is available. Some studies suggest that the hormones ghrelin and LEAP2, which act as agonist and antagonist/inverse agonist, respectively, for the G protein coupled receptor GHSR, may influence the development of MAFLD. For instance, ghrelin increases hepatic fat whereas synthetic GHSR antagonists do the opposite. Also, hepatic steatosis is less prominent in standard chow-fed ghrelin-KO mice but more prominent in 42% high-fat diet-fed female LEAP2-KO mice. MethodsHere, we sought to determine the therapeutic potential of a long-acting LEAP2 analog (LA-LEAP2) to treat MAFLD in mice. LEAP2-KO and wild-type littermate mice were fed a Gubra-Amylin-NASH (GAN) diet for 10 or 40 wks, with some randomized to an additional 28 or 10 days of GAN diet, respectively, while treated with LA-LEAP2 vs Vehicle. Various metabolic parameters were followed and biochemical and histological assessments of MAFLD were made. ResultsAmong the most notable metabolic effects, daily LA-LEAP2 administration to both LEAP2-KO and wild-type littermates during the final 4 wks of a 14 wk-long GAN diet challenge markedly reduced liver weight, hepatic triglycerides, plasma ALT, hepatic microvesicular steatosis, hepatic lobular inflammation, NASH activity scores, and prevalence of higher-grade fibrosis. These changes were accompanied by prominent reductions in body weight, without effects on food intake, and reduced plasma total cholesterol. Daily LA-LEAP2 administration during the final 10 d of a 41.5 wk-long GAN diet challenge also reduced body weight, plasma ALT, and plasma total cholesterol in LEAP2-KO and wild-type littermates and prevalence of higher grade fibrosis in LEAP2-KO mice. ConclusionsAdministration of LA-LEAP2 to mice fed a MAFLD-prone diet markedly improves several facets of MAFLD, including hepatic steatosis, hepatic lobular inflammation, higher-grade hepatic fibrosis, and transaminitis. These changes are accompanied by prominent reductions in body weight and lowered plasma total cholesterol. Taken together, these data suggest that LEAP2 analogs such as LA-LEAP2 hold promise for the treatment of MAFLD and obesity.

Benefit of Early Ruxolitinib Initiation Regardless of Fibrosis Grade in Patients with Primary Myelofibrosis: A Post Hoc Analysis of the Single-Arm Phase 3b JUMP Study.

Bone marrow fibrosis (BMF) is an adverse prognostic factor for myelofibrosis (MF). The single-arm, open-label, phase 3b JUMP trial (NCT01493414) assessed the safety and efficacy of the JAK1/JAK2 inhibitor ruxolitinib in patients with symptomatic MF. This post hoc analysis investigated the impact of BMF grade on response and outcomes in patients with primary MF (PMF) from the JUMP study. BMF was assessed by biopsy and graded from 0 to 3; grades 0-1 were considered low-grade fibrosis (LGF) and grades 2-3 were considered high-grade fibrosis (HGF). Patients with LGF (n = 268) had lower rates of cytopenias at baseline but showed comparable disease burden vs. patients with HGF (n = 852). The proportion of patients achieving a spleen response was greater in the LGF group vs. the HGF group at Week 24 and at any time during the study, while overall survival estimates were improved in patients with LGF vs. patients with HGF. Early initiation of ruxolitinib therapy (within 2 years of diagnosis) was associated with increased response rates in all patients. These results highlight the efficacy of ruxolitinib in symptomatic patients with PMF, with the greatest clinical improvements observed in patients with LGF and in patients who received early treatment.

Native liver survival in bile salt export pump deficiency: results of a retrospective cohort study.

Bile salt export pump (ABCB11) deficiency [Progressive familial intrahepatic cholestasis (PFIC2)] is the most common genetic cause of PFIC and is associated with pruritus and progressive liver disease. Surgical biliary diversion or pharmacological [ileal bile acid transporter inhibitor (IBATi)] approaches can be used to block the recirculation of bile acids to the liver. There is a paucity of detailed data on the natural history and, in particular, the longitudinal evolution of bile acid levels to predict treatment response. Cross-sectional data from large international consortia suggested a maximum cutoff value of bile acids after the intervention to predict a successful outcome. This retrospective, single-center, cohort study included all patients with confirmed biallelic pathogenic ABCB11 genotype PFIC2 treated at our institution with ≥2 years follow-up. The outcomes of interventions and predictors of long-term health were analyzed. Forty-eight cases were identified with PFIC2. Eighteen received partial external biliary diversion (PEBD) surgery, and 22 patients underwent liver transplantation. Two patients developed HCC and 2 died. Improved survival with native liver was closely associated with genotype, complete normalization of serum bile acids following PEBD, and alleviation of pruritus. Persistence of mild-to-moderate elevation of bile acids or a secondary rise following normalization was associated with liver disease progression and led to transplantation, suggesting that any prolonged elevation of bile acids worsens the chance of native liver survival. Higher-grade fibrosis at the time of PEBD was not associated with reduced long-term native liver survival. Patients with PFIC2 benefit from PEBD even at a stage of advanced fibrosis. Serum bile acid levels are an early predictor of treatment response and might serve as the gold standard in the evaluation of novel therapies including IBATi.

Hepatocellular carcinomas, exhibiting intratumor fibrosis, express cancer-specific extracellular matrix remodeling and WNT/TGFB signatures, associated with poor outcome.

HCC, the third leading cause of cancer-related death, arises in the context of liver fibrosis. Although HCC is generally poorly fibrogenic, some tumors harbor focal intratumor extracellular matrix (ECM) deposits called "fibrous nests." To date, the molecular composition and clinical relevance of these ECM deposits have not been fully defined. We performed quantitative matrisome analysis by tandem mass tags mass spectrometry in 20 human cancer specific matrisome (HCCs) with high or low-grade intratumor fibrosis and matched nontumor tissues, as well as in 12 livers from mice treated with vehicle, carbon tetrachloride, or diethylnitrosamine. We found 94 ECM proteins differentially abundant between high and low-grade fibrous nests, including interstitial and basement membrane components, such as several collagens, glycoproteins, proteoglycans, enzymes involved in ECM stabilization and degradation, and growth factors. Pathway analysis revealed a metabolic switch in high-grade fibrosis, with enhanced glycolysis and decreased oxidative phosphorylation. Integrating the quantitative proteomics with transcriptomics from HCCs and nontumor livers (n = 2,285 samples), we identified a subgroup of fibrous nest HCCs, characterized by cancer-specific ECM remodeling, expression of the WNT/TGFB (S1) subclass signature, and poor patient outcome. Fibrous nest HCCs abundantly expressed an 11-fibrous-nest - protein signature, associated with poor patient outcome, by multivariate Cox analysis, and validated by multiplex immunohistochemistry. Matrisome analysis highlighted cancer-specific ECM deposits, typical of the WNT/TGFB HCC subclass, associated with poor patient outcomes. Hence, histologic reporting of intratumor fibrosis in HCC is of clinical relevance.

Human Neutrophil α-Defensins 1-3 Are Upregulated in the Microenvironment of Fibrotic Liver.

Background and Objectives: Neutrophil infiltration is an established signature of Non-Alcoholic Fatty Liver Disease (NAFLD) and Steatohepatitis (NASH). The most abundant neutrophilic peptide, alpha-defensin, is considered a new evolving risk factor in the inflammatory milieu, intimately involved in lipid mobilization. Our objective is to assess for potential association between alpha-defensin immunostains and NAFLD severity. Materials and Methods: We retrospectively investigated the liver biopsies of NAFLD/NASH patients, obtained at Hillel Yaffe Medical center between the years 2012 and 2016. Patients' characteristics were recorded, including relevant blood tests at the time of biopsy. Each biopsy was semi-quantitatively scored using NAFLD Activity Score (NAS) and NASH fibrosis stage. The biopsies were immunostained for alpha-defensin. The precipitation of alpha-defensin was correlated to NAS and fibrosis. Results: A total of 80 biopsies were evaluated: male ratio 53.2%, mean age 44.9 ± 13.2 years, 54 had fibrosis grades 0-2, and 26 were grade 3-4. Conventional metabolic risk factors were more frequent in the high-grade fibrosis group. Immunostaining for alpha-defensin disclosed higher intensity (a.u.) in grade 3-4 fibrosis relative to grades 0-2, 25% vs. 6.5%, p < 0.05, respectively. Moreover, alpha-defensin staining was nicely co-localized with fibrosis. Conclusions: In our group of NASH/NAFLD patients, higher metabolic risk profile was associated with higher fibrosis grade. Immunostaining for alpha-defensin showed patchy intense staining concordant with high fibrosis, nicely co-localized with histological fibrosis. Whether alpha-defensin is a profibrotic risk factor or merely risk marker for fibrosis must be clarified in future studies.

A Unique Immune-Related Gene Signature Represents Advanced Liver Fibrosis and Reveals Potential Therapeutic Targets.

Innate and adaptive immune responses are critically associated with the progression of fibrosis in chronic liver diseases. In this study, we aim to identify a unique immune-related gene signature representing advanced liver fibrosis and to reveal potential therapeutic targets. Seventy-seven snap-frozen liver tissues with various chronic liver diseases at different fibrosis stages (1: n = 12, 2: n = 12, 3: n = 25, 4: n = 28) were subjected to expression analyses. Gene expression analysis was performed using the nCounter PanCancer Immune Profiling Panel (NanoString Technologies, Seattle, WA, USA). Biological meta-analysis was performed using the CBS Probe PINGSTM (CbsBioscience, Daejeon, Korea). Using non-tumor tissues from surgically resected specimens, we identified the immune-related, five-gene signature (CHIT1_FCER1G_OSM_VEGFA_ZAP70) that reliably differentiated patients with low- (F1 and F2) and high-grade fibrosis (F3 and F4; accuracy = 94.8%, specificity = 91.7%, sensitivity = 96.23%). The signature was independent of all pathological and clinical features and was independently associated with high-grade fibrosis using multivariate analysis. Among these genes, the expression of inflammation-associated FCER1G, OSM, VEGFA, and ZAP70 was lower in high-grade fibrosis than in low-grade fibrosis, whereas CHIT1 expression, which is associated with fibrogenic activity of macrophages, was higher in high-grade fibrosis. Meta-analysis revealed that STAT3, a potential druggable target, highly interacts with the five-gene signature. Overall, we identified an immune gene signature that reliably predicts advanced fibrosis in chronic liver disease. This signature revealed potential immune therapeutic targets to ameliorate liver fibrosis.

Correlation between adiponectin level and the degree of fibrosis in patients with NAFLD

Abstract Background Non-alcoholic Fatty Liver Disease (NAFLD) is one of the most prevalent chronic liver diseases particularly in Egypt. It is defined as accumulation of lipids inside the hepatocytes, in absence of other etiologies of hepatic damage. It is frequently associated with obesity, diabetes mellitus and metabolic syndrome. Objective To find out the correlation between the degree of liver fibrosis in Non-alcoholic Fatty Liver Disease patients and their serum Adiponectin level as a future non-invasive method for assessment of liver fibrosis to substitute liver biopsy to avoid its hazardous complication. Also to study the correlation between diabetes mellitus as well as obesity and serum Adiponetctin level. Patients and Methods 50 patients were selected to participate in our study based on our inclusion criteria. They were recruited from the Internal Medicine department, Gastro-intestinal clinic in AlDemerdash Hospital using a convenient sampling method. Diagnoses of NAFLD (Non-alcoholic fatty liver disease) was confirmed by laboratory markers (AST, ALT, Lipid profile), ultrasound as well as fibroscan examination. Results Analyzing adiponectin levels showed that -besides its significant correlation with BMI, hypertension, diabetes mellitus and dyslipidemia- it was significantly lower in high grade fibrosis group compared to low grade fibrosis group with P-value of (0.000) and a cutoff value for stage 3/4 fibrosis of about 2.31μg/ml which marked a promising hope of adeponictin being of protective value against liver fibrosis. However, more studies performed on populations of different sizes and characteristics are recommended to allow more accurate generalization of the results and hopefully exploring a new horizon for the follow up and treatment of patients with chronic liver disease especially NAFLD. Conclusion Adiponectin is an abundant adipocyte-derived protein with well-established antiatherogenic, insulin-sensitizing and anti-inflammatory properties. The liver is a major target organ for adiponectin especially in fatty liver diseases and this adipocytokine has the ability to control many liver functions including metabolism, inflammation and fibrosis. Both serum levels and hepatic adiponectin receptor expression are decreased in NAFLD. Therefore, either adiponectin itself or adiponectin-inducing agents might be of key therapeutic interest in the near future in the treatment of NAFLD.

Correlation between adiponectin level and the degree of fibrosis in patients with non-alcoholic fatty liver disease

BackgroundNon-alcoholic fatty liver disease (NAFLD) is one of the most prevalent chronic liver diseases, particularly in Egypt. It is defined as the accumulation of lipids inside the hepatocytes, in the absence of other etiologies of hepatic damage. It is frequently associated with obesity, diabetes mellitus, and metabolic syndrome. Adiponectin is an abundant adipocyte-derived protein with well-established anti-atherogenic, insulin-sensitizing, and anti-inflammatory properties. The liver is a major target organ for adiponectin especially in fatty liver diseases, and this adipocytokine has the ability to control many liver functions including metabolism, inflammation, and fibrosis. In this study, we aimed to find out the correlation between the degree of liver fibrosis in NAFLD patients and their serum adiponectin level as a future non-invasive method for the assessment of liver fibrosis to substitute liver biopsy to avoid its hazardous complication and also to study the correlation between diabetes mellitus as well as obesity and serum adiponectin level.ResultsFifty patients were selected to participate in our study based on our inclusion criteria. They were recruited from the Internal Medicine Department, Gastroenterology Clinic in Al-Demerdash Hospital using a convenient sampling method. Diagnosis of NAFLD was confirmed by laboratory markers: aspartate aminotransferase (AST), alanine aminotransferase (ALT), lipid profile, ultrasound, and FibroScan examination. Analyzing the adiponectin levels showed that besides its significant correlation with body mass index (BMI), hypertension, diabetes mellitus, and dyslipidemia, it was significantly lower in the high-grade fibrosis group compared to the low-grade fibrosis group with a P-value of (0.000) and a cutoff value for stage 3/4 fibrosis of about 2.31 μg/ml which marked a promising hope of adiponectin being of protective value against liver fibrosis.ConclusionBoth serum levels and hepatic adiponectin receptor expression are decreased in NAFLD. Therefore, either adiponectin itself or adiponectin-inducing agents might be of key therapeutic interest in the near future in the treatment of NAFLD.

Outcomes of Patients with Baseline Marrow Fibrosis at Allogeneic Hematopoietic Cell Transplantation

Introduction:Allogeneic hematopoietic cell transplantation (allo-HCT) is a curative option for several relapsed and refractory hematologic malignancies, including acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). Success of allo-HCT lies in complete engraftment of donor hematopoietic cells and reconstitution of donor hematopoiesis. Among the well-defined factors of poor engraftment include HLA disparity between recipient and donor, T-cell-depleted grafts, reduced-intensity conditioning, and low stem cell dose. However, bone marrow reticulin fibrosis (MF) was not clearly defined as a risk factor.Marrow fibrosis has not been observed to be associated with impaired engraftment in primary myelofibrosis patients undergoing allo-HCT. However, in patients with MDS a pre-existent MF showed a poor engraftment and secondary graft failure; Particularly, in patients with higher WHO grade fibrosis tend to have poor donor chimerism and transfusion dependency. But, this was not associated with any adverse effect on survival. Further studies have showed the time to neutrophil engraftment is prolonged in those with higher degree of MF but not in the lower grades. Studies evaluating these findings in AML with marrow fibrosis are lacking.The current study is aimed at estimating the incidence of poor engraftment in AML/MDS patients with MF who underwent allo-HCT at our institution and to compare the incidence between those with and without MF.Methods:We retrospectively reviewed all AML/MDS patients that underwent allo-HCT between 2006 and 2015 at Washington University, St Louis. We included patients aged >18, and that underwent first allo-HCT. Patients that underwent second or later allo-HCT were excluded. Pre-transplantation bone marrows biopsies were evaluated for evidence of reticulin fibrosis and WHO grade (low grade/0-1, high grade/2-3) prior to allo-HCT. Primary end point was complete donor engraftment status measured using PCR based single-tandem repeats at day 30 post-allo-HCT. Secondary end point was to evaluate neutrophil engraftment, absolute platelet engraftment, day 100 engraftment status and survival outcomes. Neutrophil and platelet engraftment were defined per standard CIBMTR criteria.The association between primary outcome and clinical risk factors was examine via univariate and multivariate logistic regression. The associations between neutrophil/platelet engraftment and clinical risk factors were examined via univariate and multivariate general linear modeling. The relationship between marrow fibrosis grade and overall survival, as well as competing risk survival of relapse and non-relapse mortality, was investigated by Cox proportional hazard regression.Results:Seven hundred and twenty-nine patients were included in the final analysis (grade 0/1 n=662; grade2/3 n=67) (Table 1). Patients in higher-grade fibrosis are relatively older (61 vs 56 years) are more likely to have MDS, likely to have active disease at transplantation (55% vs. 31%), and have had reduced intensity conditioning. Compared to patients with completed engraftment, patients in no-engraftment group are older (61.3 vs. 55.5; p= <0.0001) and a greater number of patients in that group have a higher grade of fibrosis (16% vs. 8%; p= 0.0064). Univariate analysis shows higher grade of MF is associated with engraftment failure at day 30 (odds ratio 2.247 95%CI 1.24-4.07; p= 0.008). However, in multivariate analysis, this significance was lost (odds ratio 1.77; CI 0.90-3.50; p= 0.10). MF grade was not associated with neutrophil and platelet engraftment, engraftment status at 100 days, overall survival, relapse free survival and non-relapse mortality. Factors associated with completed engraftment at day 30 post-allo-HCT are summarized in Table 2.Conclusions:Marrow fibrosis preceding allo-HCT was not associated with engraftment failure at day 30 or after transplantation with the adjustment for other risk factors.MF was not associated with other engraftment parameters or survival. Further analyses evaluating the effect of resolution of MF on survival outcomes is underway. It is imperative that several disease and transplantation characteristics play significant role than MF itself in engraftment failure supporting the theory that engraftment is an immunologic function. [Display omitted] DisclosuresVij:Amgen: Honoraria, Research Funding; Abbvie: Honoraria; Takeda: Honoraria, Research Funding; Konypharma: Honoraria; Janssen: Honoraria; Bristol-Meyers-Squibb: Honoraria; Jazz: Honoraria; Celgene: Honoraria.

Transient elastography correlated to four different histological fibrosis scores in children with liver disease

Currently, liver histology is the gold standard for the detection of liver fibrosis. In recent years, new methods such as transient elastography (TE) have been introduced into clinical practice, which allow a non-invasive assessment of liver fibrosis. The aim of the present study was to investigate the predictive value of TE for higher grade fibrosis and whether there is any relevance which histologic score is used for matching. For this purpose, we compared TE with 4 different histologic scores in pediatric patients with hepatopathies. Furthermore, we also determined the aspartate aminotransferase-to-platelet ratio (APRI) score, another non-invasive method, to investigate whether it is equally informative. Therefore, liver fibrosis in 75 children was evaluated by liver biopsy, TE and laboratory values. Liver biopsies were evaluated using four common histological scoring systems (Desmet, Metavir, Ishak and Chevalier’s semi-quantitative scoring system). The median age of the patients was 12.3 years. TE showed a good correlation to the degree of fibrosis severity independent of the histological scoring system used. The accuracy of the TE to distinguish between no/minimal fibrosis and severe fibrosis/cirrhosis was good (p = 0.001, AUC-ROCs > 0.81). The optimal cut-off value for the prediction of severe fibrosis was 10.6 kPa. In contrast, the APRI score in our collective showed no correlation to fibrosis.Conclusion: TE shows a good correlation to the histological findings in children with hepatopathy, independent of the used histological scoring system.What is Known:• The current gold standard for detecting liver fibrosis is liver biopsy. Novel non-invasive ultrasound-based methods are introduced to clinical diagnostics.• Most histological scores have been developed and evaluated in adult populations and for only one specific liver disease.What is New:• Transient elastography (TE) in children showed a good correlation to fibrosis severity irrespective of the utilized histological scoring system.• The aspartate aminotransferase-to-platelet ratio (APRI) showed no correlation with different stages of liver fibrosis in children.

Does liver biopsy accurately measure fibrosis in Fontan-associated liver disease? A comparison of liver biopsy pre-combined heart and liver transplant and liver explant post-transplant.

The accuracy of liver biopsy to stage fibrosis due to Fontan-associated liver disease (FALD) remains unclear. We compared the results of biopsy pre-combined heart and liver transplantation (CHLT) to the results of whole liver explant. Liver biopsy and explants from 15 Fontan patients (ages 16-49, median 28years) were retrospectively reviewed. Staging was as follows: stage 0: no fibrosis, stage 1: pericellular fibrosis, stage 2: bridging fibrosis, and stage 3: regenerative nodules. There is no stage 4. Clinical characteristics including Model of End-stage Liver Disease eXcluding INR and Varices, Ascites, Splenomegaly, and Thrombocytopenia (VAST) scores were collected, and descriptive statistics and Mann-Whitney U tests were used to analyze the data. All patients had biopsies with at least bridging fibrosis, and all had nodularity on explant; transjugular biopsy never overestimated fibrosis. Explant showed higher-grade fibrosis (stage 3) than pre-CHLT biopsy (stage 2) in 6 of 15 patients and equal grade of fibrosis (stage 3) in 9 of 15 patients. Though clinical characteristics varied significantly, VAST score was ≥2 in all but two patients. Transjugular liver biopsy does not overestimate and can underestimate fibrosis in Fontan patients undergoing CHLT, likely due to the patchy nature of fibrosis in FALD.

Anti-fibrotic effect of intravenous umbilical cord-derived mesenchymal stem cells (UC-MSCs) injection in experimental rats induced liver fibrosis.

Aim To investigate the effect of umbilical cord-derived mesenchymal stem cells (UC-MSCs) administration among liver fibrosis experimental rat model via the regulation of angiotensin II type 1 receptor (AT1R) and platelet-derived growth factor-β (PDGF-β) due to their therapeutic potential to replace liver transplantation for advanced liver fibrosis. Yet the mechanism of action has been questionably associated with UC-MSCs fibrosis regression properties. Methods Sprague-Dawley (SD) rats (n=18) were separated into three groups (control, untreated liver fibrosis, and UC-MSCs treated group). Serum PDGF-β level was determined by enzymelinked immunosorbent assay (ELISA) following 14 days of UCMSCs injection. Meanwhile, AT1R expression was interpreted based on immunoreactive score (IRS) stained using polyclonal antibody and liver fibrosis stained with hematoxylin & eosin was graded using the METAVIR score. Results UC-MSCs were isolated successfully from rat umbilical cord. Liver fibrosis was observed following 14 weeks of CCl4 injection concurrent with higher serum level of PDGF-β, but the UC-MSCs-treated group had lower level (980.08 ±289.41 and 606.42±109.85 for untreated liver fibrosis and UC-MSCs treated group, respectively; p=0.004). There was also a high expression of AT1R among untreated liver fibrosis group, as well as highgrade liver fibrosis versus localized fibrosis and low level of AT1R expression among UC-MSCs treated-group (p=0.001). Conclusion UC-MSCs administration could ameliorate liver fibrosis by reducing the AT1R expression and PDGF-β serum levels, and intervention through this signaling pathway could be alternative evidence for the causative of positive outcome.

Three-dimensional CT texture analysis of anatomic liver segments can differentiate between low-grade and high-grade fibrosis

BackgroundCT texture analysis (CTTA) has been successfully used to assess tissue heterogeneity in multiple diseases. The purpose of this work is to demonstrate the value of three-dimensional CTTA in the evaluation of diffuse liver disease. We aimed to develop CTTA based prediction models, which can be used for staging of fibrosis in different anatomic liver segments irrespective of variations in scanning parameters.MethodsWe retrospectively collected CT scans of thirty-two chronic hepatitis patients with liver fibrosis. The CT examinations were performed on either a 16- or a 64-slice scanner. Altogether 354 anatomic liver segments were manually highlighted on portal venous phase images, and 1117 three-dimensional texture parameters were calculated from each segment. The segments were divided between groups of low-grade and high-grade fibrosis using shear-wave elastography. The highly-correlated features (Pearson r > 0.95) were filtered out, and the remaining 453 features were normalized and used in a classification with k-means and hierarchical cluster analysis. The segments were split between the train and test sets in equal proportion (analysis I) or based on the scanner type (analysis II) into 64-slice train 16-slice validation cohorts for machine learning classification, and a subset of highly prognostic features was selected with recursive feature elimination.ResultsA classification with k-means and hierarchical cluster analysis divided segments into four main clusters. The average CT density was significantly higher in cluster-4 (110 HU ± SD = 10.1HU) compared to the other clusters (c1: 96.1 HU ± SD = 11.3HU; p < 0.0001; c2: 90.8 HU ± SD = 16.8HU; p < 0.0001; c3: 93.1 HU ± SD = 17.5HU; p < 0.0001); but there was no difference in liver stiffness or scanner type among the clusters. The optimized random forest classifier was able to distinguish between low-grade and high-grade fibrosis with excellent cross-validated accuracy in both the first and second analysis (AUC = 0.90, CI = 0.85–0.95 vs. AUC = 0.88, CI = 0.84–0.91). The final support vector machine model achieved an excellent prediction rate in the second analysis (AUC = 0.91, CI = 0.88–0.94) and an acceptable prediction rate in the first analysis (AUC = 0.76, CI = 0.67–0.84).ConclusionsIn conclusion, CTTA-based models can be successfully applied to differentiate high-grade from low-grade fibrosis irrespective of the imaging platform. Thus, CTTA may be useful in the non-invasive prognostication of patients with chronic liver disease.

MPN-402: Delayed Resolution of Bone Marrow Morphological Changes of Myelofibrosis Following Successful Stem Cell Transplant and Molecular Clearance of Disease: A Cautionary Tale

Context: Allogeneic stem cell transplant (allo-SCT) is a curative option for patients with myelofibrosis (MF). Reversal of the bone marrow (BM) morphologic features of MF often lags following allo-SCT and may create diagnostic challenges in disease surveillance. Objective: To determine which morphologic parameters could determine the disease persistence in patients with BM fibrosis following allo-SCT. Design: We compared the morphologic features typically associated with myelofibrosis in pre-transplant and post-transplant BM samples in a cohort of patients with BM fibrosis who achieved complete clearance of JAK2, MPL, and/or CALR mutations following allo-SCT. Setting: MDACC. Patients or other participants: Patients with primary or secondary MF and a driver mutation in a pre-transplant BM sample who achieved a molecular clearance after allo-SCT. Interventions: N/A. Main outcome measures: N/A. Results: Patients (n=27) included 12 men and 15 women with a median age of 63 years (range 42-76 years) diagnosed with primary myelofibrosis (n=17), post-essential thrombocythemia (post-ET) MF (n=5), and post-polycythemia vera (post-PV) MF (n=5). Driver mutations included JAK2 p.V617F (n=18), MPL (n=4), and CALR (n=7). Two patients with post-PV-MF had concurrent JAK2 and MPL mutations. Compared to pre-transplant samples, post-transplant samples demonstrated significantly fewer megakaryocytes (p Conclusion: Reversal of BM morphologic features following allo-SCT for MF lags behind molecular clearance. Re-appearance of erythroid islands, normalization of BM cellularity and disappearance of high-grade BM fibrosis and endosteal localization of megakaryocytes represent reassuring findings in evaluation of a response or recurrence.

Hybrid precutting EMR-EFTR resection for a tricky rectal adenoma recurrence with high-grade fibrosis

Hybrid precutting EMR-EFTR resection for a tricky rectal adenoma recurrence with high-grade fibrosis

P1.43: Use of transient elastography to determine liver fibrosis in pediatric intestinal failure

Introduction: Intestinal failure associated liver disease (IFALD) is a significant complication in pediatric intestinal failure. Liver fibrosis with or without cholestasis is a frequent occurrence in IFALD. However, the diagnosis of fibrosis in IFALD is challenging and currently limited to liver biopsy. Our objective was to evaluate the use of transient elastography (TE) in pediatric IF patients to determine its efficacy for monitoring liver fibrosis as an alternative to liver biopsy. Methods: A retrospective cohort study of IF patients between January 1, 2015 to December 31, 2017. The study cohort included a sample of patients who had a routine liver biopsy during an operative procedure and completed transient elastography at the same time. Liver biopsies were evaluated for stage of fibrosis using the modified Scheuer score. Statistical analysis included univariate analysis and Chi-square methods. High (grade 3–4) versus low (grade 0–2) fibrosis and TE scores were compared to determine sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). Receiver operating characteristic (ROC) curves were also used to determine the ability of TE to discriminate various stages of fibrosis. Data is presented as medians with interquartile ranges and frequencies. Results: 30 patients (21 male [70%]) with a median age at biopsy of 320 days (154–1776) were evaluated. Majority of patients had an etiology of abdominal wall defects (12[40%]) and necrotizing enterocolitis (9[30%]). Median length of PN therapy was 190 days (120–789). All TE measurements were completed within a median of 8.5 days (5.0–13.3) of the liver biopsy. Individuals with histologic fibrosis scores of 0–1 (n=18) had a median TE score of 4.9 (3.9–7.1), fibrosis scores of 2 (n=5) had a median TE of 6.7 (4.4–11.4) and those with a fibrosis score of 3 (n=7) had a median TE of 12.1 (6.3–14.6). No patients had grade 4 fibrosis. Dichotomous evaluation of the fibrosis scores (low vs high grade) compared to the chronic cholestatic scale for Fibroscan demonstrated 95.7% sensitivity, 57.1% specificity, 88.0% PPV and 80.0% NPV for low grade fibrosis. The c-statistic for the ROC curve was 0.764. Conclusion: Fibrosis in patients with IF is often difficult to assess without liver biopsy. TE has only been evaluated in small cohorts but shows potential as a non-invasive method to monitor low vs high grade fibrosis in children with IF. Further studies in a larger cohort are required.

Hippo Kinase Inactivation Contributes to Del(20q) Malignancies and Cooperates with JAK2-V617F to Promote Myelofibrosis in Mice

Recurring chromosome abnormalities are frequent events in cancer and are especially prevalent in hematologic neoplasms. Somatic heterozygous deletions on chromosome 20q are detected in a variety of hematopoietic malignancies including myelodysplastic syndrome (MDS), classical myeloproliferative neoplasm (MPN), MDS/MPN overlap disorders such as chronic myelomonocytic leukemia (CMML), and acute leukemias. Del(20q) is especially prevalent in MPN patients (~10-15%), where it is the most commonly detected cytogenetic abnormality associated with primary myelofibrosis (PMF) and post-polycythemia vera myelofibrosis (MF). This suggests that heterozygous loss of genes in the del(20q) common deleted region (CDR) may contribute to adverse MPN progression. Despite these observations, relatively few genes located within the CDR have been unambiguously implicated, highlighting a significant need for further investigation. To identify genes that may play an important role in the biology of del(20q)-associated malignancies we utilized a published gene expression dataset of bone-marrow derived CD34+ cells from MDS patients and healthy controls (Gerstung et al, 2015). Comparison of the patients harboring del(20q) to healthy controls revealed STK4 (encoding Hippo kinase MST1) to be the most significantly downregulated gene (mean: 3.5-fold) among those located within the chromosome 20q CDR. We therefore set out to assess the role of Hippo kinase inactivation in hematologic malignancy using conditional gene inactivation in mice.We found that complete inactivation of both Hippo kinases (Stk4 and Stk3) within the hematopoietic system using Vav1-Cre (Stk4-/-Stk3-/-) resulted in a lethal bone marrow failure (median survival: 7 weeks) associated with myelodysplastic features and frequent extramedullary hematopoiesis in the spleen. A single copy of Stk4 rescued the lethality due to bone marrow failure, however sub-haploinsufficient mice displayed thrombocytopenia with a trend towards mild anemia; phenotypes that closely resemble those observed in MDS patients with isolated del(20q). Both a reduced number of mature megakaryocytes and the presence of dysplastic megakaryocytes were apparent in bone marrow sections. Inducible Hippo kinase inactivation in adult mice using the Mx1-Cre system similarly recapitulated several phenotypic features of both MDS and MPN. In competitive bone marrow transplant assays we found that Stk4-/-Stk3-/- hematopoietic stem cells (HSC) completely lacked engraftment potential and failed to reconstitute normal hematopoiesis, revealing a potential role for Hippo kinase function in HSC homing and retention in the bone marrow. Heterozygous HSCs maintained relatively normal steady-state hematopoiesis in peripheral blood and bone marrow for up to 48 weeks in primary and secondary transplantations, although upon aging these mice were prone to development of thrombocytopenia with increased mean platelet volume.Given the high frequency of del(20q) in MPN, especially PMF, we asked whether heterozygous Hippo kinase inactivation may cooperate with the common driver mutation JAK2-V617F to accelerate disease progression. Using an HSC-enriched retroviral transduction/transplantation model in C57BL/6 recipient mice, we monitored MPN progression for 36 weeks in heterozygous Stk4+/-Stk3+/-, or control (Vav1-Cre-), cells with or without expression of JAK2-V617F. While both JAK2-V617F groups initially displayed a similar degree of polycythemia relative to controls, we found heterozygous Hippo kinase inactivation to promote accelerated disease progression towards lethal bone marrow fibrosis during the course of observation. Recipients in this group showed significantly reduced overall survival, which was associated with higher grade fibrosis in bone marrow, elevated peripheral granulocyte counts, enhanced splenomegaly, and increased frequencies of hematopoietic stem and progenitor populations in the spleen. Together, these findings implicate aberrant Hippo kinase loss-of-function in the pathogenesis of del(20q)-associated hematologic malignancies, and shed new light on the molecular events that contribute to adverse MPN progression. DisclosuresBejar:Genoptix: Consultancy; Modus Outcomes: Consultancy; Celgene: Consultancy, Honoraria; Takeda: Research Funding; Astex/Otsuka: Consultancy, Honoraria; AbbVie/Genentech: Consultancy, Honoraria; Foundation Medicine: Consultancy. Guan:Vivace: Equity Ownership.

Reduced Intensity Allogeneic Stem Cell Transplantation for Younger Patients with Myelofibrosis

INTRODUCTION Allogeneic stem cell transplantation (alloSCT) is the only curative procedure for primary and secondary myelofibrosis (PMF, SMF). Elderly people are mainly affected, limiting the feasibility of intensive myeloablative chemotherapy regimens. The introduction of reduced-intensity conditioning (RIC) made alloSCT feasible and effective for old patients. Nevertheless, the incidence of PMF and SMF is not negligible in young patients, theoretically able to tolerate also high-intensity therapy. Very few data are available about the efficacy of RIC-alloSCT in the particular setting of young-aged MF patients.PATIENTS AND METHODS This study includes 56consecutive Myelofibrosis young patients (age <55y) who received alloSCT allogeneic stem cell procedure between 2002 and 2016 at the University Hospital Hamburg/Germany treated in UK-Eppendorf, Hamburg. Only 4 patients were previously splenectomized. Patients mostly fall into intermediate risk groups according to DIPSS model (int-1: 22; int-2: 24; high: 3). Four patients belonged to the high-risk triple-negative category (JAK2/CALR/MPL-). ASXL1 additional mutation was tested in 50 patients (, and positive: in 17) cases. In 96% graft source was peripheral blood stem cells, 2 patients received bone marrow stem cell. Only 30% of patients had a 10/10 HLA-matched sibling donor, the others were transplanted from fully-matched (36%) or partially-matched (34%) unrelated donor. All transplants were conditioned according the EBMT protocol with Busulfan (10 mg/Kg PO or 8 mg/Kg IV), Fludarabine (150 mg/m2), ATLG (Grafalon® Neovii, Germany) administered in 3 days at a dose of 20 mg/Kg die for MUD, 10 mg/Kg die for MRD transplants, followed by Cylosporin A, and Mycophenolate in the first 28 days was added only in MUD/MMUD transplants.RESULTS Engraftment rate was 98%, with a median neutrophil engraftment time of 15 days. Platelet engraftment was reached by 51 patients (91%) in a median time of 19 days. Four patients (7%) developed poor graft function, successfully treated with CD34+ selected PBSC boost. After a median follow up of 8.6 y estimated 5y PFS and OS was 68% and 82% respectively. DIPSS risk and donor HLA-matching resulted the only significant impacting factors on OS. Neither cytogenetic nor molecular abnormalities (included ASXL1) were significantly related to OS. Twenty-five patients (44%) experienced acute GVHD grade >1. Chronic GVHD was observed in 34 patients (65%), mostly (82%) beginning in the first 300 days after transplantation. Cumulative incidence of TRM was 7% at 1 year, with a plateau after the first year (5y TRM = 12%). TRM was observed only in patients with maximal grade (3) of marrow fibrosis (p=0.00). Furthermore, TRM never occurred in previously splenectomized patients (p=0.00), but no significant impact from splenectomy on OS was observed (p=0.32). After transplant, 11 patients (20%) relapsed: 1 died without any treatment because of infection, 9 received DLI with durable recovery of complete remission in 3 cases, 7 patients (6 after DLI) underwent a second alloSCT, with long-term survival in 5 cases.CONCLUSION Reduced intensity conditioning followed by allogeneic SCT is a curative treatment approach for younger patients with myelofibrosis with a low NRM. The most important outcome-determining factor is donor HLA-matching. Interestingly, marrow high grade fibrosis showed to significantly impact TRM. Biological markers such as ASXL1 mutation and cytogenetics, largely known as highly predictive for poor prognosis in the disease natural course, did not show any impact on survival, suggesting that patients harboring these abnormalities could get the greatest benefit from alloSCT. According to these data, RIC-alloSCT ensures optimal engraftment and low relapse rate, being able to lead also younger MF patients to cure. Further data collection, and a prospective randomized trial are needed to confirm our conclusion. DisclosuresNo relevant conflicts of interest to declare.

Rheological determinants for simultaneous staging of hepatic fibrosis and inflammation in patients with chronic liver disease.

The purpose of this study is to investigate the use of fundamental rheological parameters as quantified by MR elastography (MRE) to measure liver fibrosis and inflammation simultaneously in humans. MRE was performed on 45 patients at 3T using a vibration frequency of 56Hz. Fibrosis and inflammation scores were obtained from liver biopsies. Biomechanical properties were quantified in terms of complex shear modulus G* as well as shear wave phase velocity c and shear wave attenuation α. A rheological fractional derivative order model was used to investigate the linear dependence of the free model parameters (dispersion slope y, intrinsic speed c0 , and intrinsic relaxation time τ) on histopathology. Leave-one-out cross-validation was then utilized to demonstrate the effectiveness of the model. The intrinsic speed c0 increases with hepatic fibrosis, while an increased relaxation time τ is reflective of more inflammation of the liver parenchyma. The dispersion slope y does not depend either on fibrosis or on inflammation. The proposed rheological model, given this specific parameterization, establishes the functional dependences of biomechanical parameters on histological fibrosis and inflammation. The leave-one-out cross-validation demonstrates that the model allows identification, from the MRE measurements, of the histology scores when grouped into low-/high-grade fibrosis and low-/high-grade inflammation with significance levels of P=0.0004 (fibrosis) and P=0.035 (inflammation). The functional dependences of intrinsic speed and relaxation time on fibrosis and inflammation, respectively, shed new light onto the impact hepatic pathological changes on liver tissue biomechanics in humans. The dispersion slope y appears to represent a structural parameter of liver parenchyma not impacted by the severity of fibrosis/inflammation present in this patient cohort. This specific parametrization of the well-established rheological fractional order model is valuable for the clinical assessment of both fibrosis and inflammation scores, going beyond the capability of the plain shear modulus measurement commonly used for MRE.