Early Stages Of Fibrosis Research Articles

Exploration of a hypoxia-immune-related microenvironment gene signature and prediction model for hepatitis C-induced early-stage fibrosis

BackgroundLiver fibrosis contributes to significant morbidity and mortality in Western nations, primarily attributed to chronic hepatitis C virus (HCV) infection. Hypoxia and immune status have been reported to be significantly correlated with the progression of liver fibrosis. The current research aimed to investigate the gene signature related to the hypoxia-immune-related microenvironment and identify potential targets for liver fibrosis.MethodSequencing data obtained from GEO were employed to assess the hypoxia and immune status of the discovery set utilizing UMAP and ESTIMATE methods. The prognostic genes were screened utilizing the LASSO model. The infiltration level of 22 types of immune cells was quantified utilizing CIBERSORT, and a prognosis-predictive model was established based on the selected genes. The model was also verified using qRT-PCR with surgical resection samples and liver failure samples RNA-sequencing data.ResultsElevated hypoxia and immune status were linked to an unfavorable prognosis in HCV-induced early-stage liver fibrosis. Increased plasma and resting NK cell infiltration were identified as a risk factor for liver fibrosis progression. Additionally, CYP1A2, CBS, GSTZ1, FOXA1, WDR72 and UHMK1 were determined as hypoxia-immune-related protective genes. The combined model effectively predicted patient prognosis. Furthermore, the preliminary validation of clinical samples supported most of the conclusions drawn from this study.ConclusionThe prognosis-predictive model developed using six hypoxia-immune-related genes effectively predicts the prognosis and progression of liver fibrosis. The current study opens new avenues for the future prediction and treatment of liver fibrosis.

P376 A prospective multi-model study on diagnostic performance of 18F-FAPI PET/CT in comparison with 18F-FDG PET/CT or magnetization transfer MR imaging to detect intestinal fibrosis in Crohn’s disease

Abstract Background Accurate and early detection of Crohn's disease (CD) induced intestinal fibrosis is crucial for effective clinical management, yet it remains an unmet requirement. The magnetization transfer MR Imaging (MTI) was reported to offer high accurancy for detection of intestinal fibrosis. Recently, the utilization of fibroblast activating protein inhibitor (FAPI) PET/CT has emerged as a promising tool for assessing fibrosis. The diagnostic efficacy of 18F-FAPI PET/CT in detecting intestinal fibrosis was compared with that of 18F-FDG PET/CT and MTI. Methods Twenty-two rats were treated with TNBS for 2-4 weeks to simulate fibrosis development, and multi-model quantitative imaging was performed during a week. The mean and maximum standardized uptake values (SUVmean and SUVmax) were calculated on 18F-FAPI and 18F-FDG PET/CT, as well as the normalized magnetization transfer ratio on MTI (normalized MTR). Pathological evaluation of intestinal fibrosis was performed, and MTI was used as the imaging standard for fibrotic analysis. Imaging and pathological criteria were used to compare the diagnostic effects of fibrosis imaging parameters. Ten patients with 34 cases of intestinal stenosis were prospectively recruited to validate their diagnostic performance using the same imaging protocol. Results In patients, the accuracy of MTI (AUCs=0.90-0.91, P≤0.05) was comparable to FAPI uptake (AUCs=0.80-0.87, both P≤0.01) but higher than that of FDG uptake in distinguishing between non-to-mild and moderate-to-severe fibrosis (AUCs=0.53-0.82, P=0.01-0.36). In rats, FAPI uptake showed a rapid response to intestinal fibrosis at an early disease phase (week 0-2) and demonstrated a significant increasing trend with prolonged treatment duration (week 0-4;). In contrast, normalized MTR and FDG uptake presented only a minimal response at week 2 and did not correlate so clearly with treatment duration. Consistently, in early disease phase (rats treated till week 2), the correlations of FAPI uptake (SUVmean: R=0.69) with fibrotic scores were stronger than those of FDG uptake (SUVmean: R=0.17) and normalized MTR (R=0.52). In late disease phase (rats treated till week 3 or 4), normalized MTR (R=0.93) showed stronger correlations with fibrotic scores than FAPI uptake (SUVmean: R=0.55) or FDG uptake (SUVmean: R=0.19). Conclusion In accurately identifying early-stage intestinal fibrosis, 18F-FAPI PET/CT demonstrates vast potential and outperforms the performance of the other two imaging modalities, warranting further research. However, 18F-FAPI PET/CT offer an equal performance for characterization of intestinal fibrosis with MTI and has significant advantages compared to 18F-FDG PET/CT in the whole disease course from Crohn's disease.

Amide Proton Transfer-Weighted Magnetic Resonance Imaging for Application in Renal Fibrosis: A Radiological-Pathological-Based Analysis

Plain Language SummaryAmide proton transfer (APT) MRI, a novel molecular MRI technique, could indirectly measure the peptide or protein concentration within tissues based on frequent exchange between amide proton and the surrounding water protons. This new technology has been used in tumor diagnosis. Given pathological conditions linked to tissue fibrogenesis are similar to carcinogenesis, we proposed the hypothesis that APT MRI can be applied to assess collagen deposition in RF. To test this, we constructed animal models for RF and enrolled CKD patients, respectively. The results suggested APT values were more sensitive to the early stage of fibrosis than ADC values, and the cortical APT was highly positively correlated with the degree of fibrosis. We further identified that the combination of two MRI protocols, cortical apparent diffusion coefficient and cortical APT, was an excellent indicator for the diagnosis of fibrosis, with much higher accuracy than a single method. To the best of our knowledge, this is the first study to explore the value of APT imaging for assessing renal interstitial fibrosis. Our findings identify APT as a novel and precise imaging diagnostic tool for future noninvasive assessment of RF. It complements with the existing imaging biomarkers and adds value to conventional MRI.

Advances in Noninvasive Molecular Imaging Probes for Liver Fibrosis Diagnosis.

Liver fibrosis is a wound-healing response to chronic liver injury, which may lead to cirrhosis and cancer. Early-stage fibrosis is reversible, and it is difficult to precisely diagnose with conventional imaging modalities such as magnetic resonance imaging, positron emission tomography, single-photon emission computed tomography, and ultrasound imaging. In contrast, probe-assisted molecular imaging offers a promising noninvasive approach to visualize early fibrosis changes invivo, thus facilitating early diagnosis and staging liver fibrosis, and even monitoring of the treatment response. Here, the most recent progress in molecular imaging technologies for liver fibrosis is updated. We start by illustrating pathogenesis for liver fibrosis, which includes capillarization of liver sinusoidal endothelial cells, cellular and molecular processes involved in inflammation and fibrogenesis, as well as processes of collagen synthesis, oxidation, and cross-linking. Furthermore, the biological targets used in molecular imaging of liver fibrosis are summarized, which are composed of receptors on hepatic stellate cells, macrophages, and even liver collagen. Notably, the focus is on insights into the advances in imaging modalities developed for liver fibrosis diagnosis and the update in the corresponding contrast agents. In addition, challenges and opportunities for future research and clinical translation of the molecular imaging modalities and the contrast agents are pointed out. We hope that this review would serve as a guide for scientists and students who are interested in liver fibrosis imaging and treatment, and as well expedite the translation of molecular imaging technologies from bench to bedside.

Metabolic profiling of murine radiation-induced lung injury with Raman spectroscopy and comparative machine learning.

Radiation-induced lung injury (RILI) is a dose-limiting toxicity for cancer patients receiving thoracic radiotherapy. As such, it is important to characterize metabolic associations with the early and late stages of RILI, namely pneumonitis and pulmonary fibrosis. Recently, Raman spectroscopy has shown utility for the differentiation of pneumonitic and fibrotic tissue states in a mouse model; however, the specific metabolite-disease associations remain relatively unexplored from a Raman perspective. This work harnesses Raman spectroscopy and supervised machine learning to investigate metabolic associations with radiation pneumonitis and pulmonary fibrosis in a mouse model. To this end, Raman spectra were collected from lung tissues of irradiated/non-irradiated C3H/HeJ and C57BL/6J mice and labelled as normal, pneumonitis, or fibrosis, based on histological assessment. Spectra were decomposed into metabolic scores via group and basis restricted non-negative matrix factorization, classified with random forest (GBR-NMF-RF), and metabolites predictive of RILI were identified. To provide comparative context, spectra were decomposed and classified via principal component analysis with random forest (PCA-RF), and full spectra were classified with a convolutional neural network (CNN), as well as logistic regression (LR). Through leave-one-mouse-out cross-validation, we observed that GBR-NMF-RF was comparable to other methods by measure of accuracy and log-loss (p > 0.10 by Mann-Whitney U test), and no methodology was dominant across all classification tasks by measure of area under the receiver operating characteristic curve. Moreover, GBR-NMF-RF results were directly interpretable and identified collagen and specific collagen precursors as top fibrosis predictors, while metabolites with immune and inflammatory functions, such as serine and histidine, were top pneumonitis predictors. Further support for GBR-NMF-RF and the identified metabolite associations with RILI was found as CNN interpretation heatmaps revealed spectral regions consistent with these metabolites.

Association between Transfusion-Related Iron Overload and Liver Fibrosis in Survivors of Pediatric Leukemia: A Cross-Sectional Study.

Patients who receive frequent blood transfusions are at an elevated risk of developing hepatic fibrosis due to iron overload in the liver. In this study, we evaluated the effectiveness of transient elastography (TE) (FibroScan®) for assessing liver fibrosis in patients with pediatric cancer. We enrolled 106 consecutive cases of acute leukemia in individuals under 21 years of age. The participants were followed for 2 years. Based on their serum ferritin (SF) levels, the patients were divided into two groups: group 1 (SF≥300 ng/mL) and group 2 (SF<300 ng/mL). A liver FibroScan® was performed, and a p-value of less than 0.05 was considered statistically significant. Among the various parameters in the liver function test (LFT), alkaline phosphatase was significantly higher in a subgroup of patients aged 5-8 years in group 2 compared to those in group 1. The indices of liver fibrosis determined by TE, including the FibroScan score, controlled attenuation parameter score, steatosis percentage, and meta-analysis of histological data in viral hepatitis score, as well as indirect serum markers of liver fibrosis such as the aminotransferase (AST)/alanine aminotransferase (ALT) ratio, Fibrosis 4 score, and AST to platelet ratio index, did not differ significantly between the two groups. The association between the TE results and LFT parameters was only significant for ALT. Transfusion-associated iron overload does not have a significant correlation with severe liver fibrosis. FibroScan® is not a sensitive tool for detecting early stages of fibrosis in survivors of pediatric leukemia.

Quantitative proteomic analysis reveals regulatory networks of extracellular matrix receptor interaction pathways in endothelial cells after myocardial infarction

Cardiac fibrosis, a significant pathological alteration following myocardial infarction (MI), remains enigmatic with respect to the role of cardiac endothelial cells (ECs). To elucidate the proteomic shifts in cardiac ECs accompanying MI-induced cardiac fibrosis, a standard MI mice model was established through ligation of the left anterior descending branch. Following 14 days of effective modeling, we isolated primary ECs from the hearts of both sham and MI models utilizing the CD31 microbeads sorting technique. Quantitative proteomics and bioinformatics methodologies, including tandem mass spectrometry, were employed to discern proteomic alterations in the primary endothelial cells of the experimental groups. Comprehensive analyses, including Gene Ontology analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, functional enrichment analysis, and functional enrichment cluster analysis, revealed an up-regulation of proteins associated with extracellular matrix-receptor interaction pathway in cardiac fibrosis post-MI. Subsequent Western blot analysis confirmed the up-regulation of specific proteins involved in this pathway, namely collagen type VI alpha 2 (Col6&amp;alpha;2), vitronectin (Vtn), and integrin beta (Itg&amp;beta;). We conclude that the expression levels of Col6&amp;alpha;2, Vtn, and Itg&amp;beta; in primary ECs during the early stage of cardiac fibrosis, 14 days post-MI, were significantly elevated compared to the sham group (P &lt; 0.05). This observation suggests that ECM-receptor interaction could potentially influence the progression of cardiac fibrosis following MI.

Utilizing MRI, [18F]FDG-PET and [89Zr]Zr-DFO-28H1 FAP-PET tracer to assess inflammation and fibrogenesis in a reproducible lung injury rat model: a multimodal imaging study.

Accurate imaging biomarkers that indicate disease progression at an early stage are highly important to enable timely mitigation of symptoms in progressive lung disease. In this context, reproducible experimental models and readouts are key. Here, we aim to show reproducibility of a lung injury rat model by inducing disease and assessing disease progression by multi-modal non-invasive imaging techniques at two different research sites. Furthermore, we evaluated the potential of fibroblast activating protein (FAP) as an imaging biomarker in the early stage of lung fibrosis. An initial lung injury rat model was set up at one research site (Lund University, Lund, Sweden) and repeated at a second site (Radboudumc, Nijmegen, The Netherlands). To induce lung injury, Sprague-Dawley rats received intratracheal instillation of bleomycin as one single dose (1,000 iU in 200 µL) or saline as control. Thereafter, longitudinal images were acquired to track inflammation in the lungs, at 1 and 2 weeks after the bleomycin challenge by magnetic resonance imaging (MRI) and [18F]FDG-PET. After the final [18F]FDG-PET scan, rats received an intravenous tracer [89Zr]Zr-DFO-28H1 (anti-FAP antibody) and were imaged at day 15 to track fibrogenesis. Upon termination, bronchoalveolar lavage (BAL) was performed to assess cell and protein concentration. Subsequently, the biodistribution of [89Zr]Zr-DFO-28H1 was measured ex vivo and the spatial distribution in lung tissue was studied by autoradiography. Lung sections were stained and fibrosis assessed using the modified Ashcroft score. Bleomycin-challenged rats showed body weight loss and increased numbers of immune cells and protein concentrations after BAL compared with control animals. The initiation and progression of the disease were reproduced at both research sites. Lung lesions in bleomycin-exposed rats were visualized by MRI and confirmed by histology. [18F]FDG uptake was higher in the lungs of bleomycin-challenged rats compared with the controls, similar to that observed in the Lund study. [89Zr]Zr-DFO-28H1 tracer uptake in the lung was increased in bleomycin-challenged rats compared with control rats (p = 0.03). Here, we demonstrate a reproducible lung injury model and monitored disease progression using conventional imaging biomarkers MRI and [18F]FDG-PET. Furthermore, we showed the first proof-of-concept of FAP imaging. This reproducible and robust animal model and imaging experimental set-up allows for future research on new therapeutics or biomarkers in lung disease.

Ex vivo tissue perturbations coupled to single-cell RNA-seq reveal multilineage cell circuit dynamics in human lung fibrogenesis.

Pulmonary fibrosis develops as a consequence of failed regeneration after injury. Analyzing mechanisms of regeneration and fibrogenesis directly in human tissue has been hampered by the lack of organotypic models and analytical techniques. In this work, we coupled ex vivo cytokine and drug perturbations of human precision-cut lung slices (hPCLS) with single-cell RNA sequencing and induced a multilineage circuit of fibrogenic cell states in hPCLS. We showed that these cell states were highly similar to the in vivo cell circuit in a multicohort lung cell atlas from patients with pulmonary fibrosis. Using micro-CT-staged patient tissues, we characterized the appearance and interaction of myofibroblasts, an ectopic endothelial cell state, and basaloid epithelial cells in the thickened alveolar septum of early-stage lung fibrosis. Induction of these states in the hPCLS model provided evidence that the basaloid cell state was derived from alveolar type 2 cells, whereas the ectopic endothelial cell state emerged from capillary cell plasticity. Cell-cell communication routes in patients were largely conserved in hPCLS, and antifibrotic drug treatments showed highly cell type-specific effects. Our work provides an experimental framework for perturbational single-cell genomics directly in human lung tissue that enables analysis of tissue homeostasis, regeneration, and pathology. We further demonstrate that hPCLS offer an avenue for scalable, high-resolution drug testing to accelerate antifibrotic drug development and translation.

Sex Drives Functional Changes in the Progression and Regression of Liver Fibrosis.

Liver fibrosis is a common and reversible feature of liver damage associated with many chronic liver diseases, and its onset is influenced by sex. In this study, we investigated the mechanisms of liver fibrosis and regeneration, focusing on understanding the mechanistic gaps between females and males. We injected increasing doses of carbon tetrachloride into female and male mice and maintained them for a washout period of eight weeks to allow for liver regeneration. We found that male mice were more prone to developing severe liver fibrosis as a consequence of early chronic liver damage, supported by the recruitment of a large number of Ly6Chigh MoMφs and neutrophils. Although prolonged liver damage exacerbated the fibrosis in mice of both sexes, activated HSCs and Ly6Chigh MoMφs were more numerous and active in the livers of female mice than those of male mice. After eight weeks of washout, only fibrotic females reported no activated HSCs, and a phenotype switching of Ly6Chigh MoMφs to anti-fibrogenic Ly6Clow MoMφs. The early stages of liver fibrosis mostly affected males rather than females, while long-term chronic liver damage was not influenced by sex, at least for liver fibrosis. Liver repair and regeneration were more efficient in females than in males.

The urinary RNA atlas of patients with chronic kidney disease

Chronic kidney disease (CKD) represents a significant global health burden. Currently employed CKD biomarkers are influenced by various factors and lack accuracy in reflecting early-stage renal fibrosis severity. Consequently, there is an urgent need for the identification of early, noninvasive CKD biomarkers. Urine, easily collectible and kidney-derived, has demonstrated potential as a diagnostic source for various kidney diseases by leveraging its RNA content. To address this, we obtained RNA-seq data pertaining to urinary RNAs from both CKD patients and healthy controls via the Gene Expression Omnibus database (GEO). The DEseq2 software was utilized to identify differentially expressed RNAs (DE-RNAs). To evaluate the overall accuracy of these DE-RNAs in urine, we performed Receiver Operating Characteristic analysis (ROC). Selected urinary RNAs were subsequently validated using reverse-transcription quantitative real-time Polymerase Chain Reaction (qRT-PCR) in conjunction with ROC analysis. Computational and experimental analyses revealed significant increases in miR-542-5p, miR-33b-5p, miR-190a-3p, miR-507, and CSAG4 within the urine of CKD patients, exhibiting high AUC values. In conclusion, our findings suggest that urinary RNAs hold promise as diagnostic biomarkers for CKD.

HMGB2 is a potential diagnostic marker and therapeutic target for liver fibrosis and cirrhosis.

High mobility group proteins 1 and 2 (HMGB1 and HMGB2) are 80% conserved in amino acid sequence. The function of HMGB1 in inflammation and fibrosis has been extensively characterized. However, an unaddressed central question is the role of HMGB2 on liver fibrosis. In this study, we provided convincing evidence that the HMGB2 expression was significantly upregulated in human liver fibrosis and cirrhosis, as well as in several mouse liver fibrosis models. The carbon tetrachloride (CCl4) induced liver fibrosis mouse model was used. AAV8-Hmgb2 was utilized to overexpress Hmgb2 in the liver, while Hmgb2-/- mice were used for loss of function experiments. The HMGB2 inhibitor inflachromene and liposome-shHMGB2 (lipo-shHMGB2) were employed for therapeutic intervention. The serum HMGB2 levels were also markedly elevated in patients with liver fibrosis and cirrhosis. Deletion of Hmgb2 in Hmgb2-/- mice or inhibition of HMGB2 in mice using a small molecule ICM slowed the progression of CCl4-induced liver fibrosis despite constant HMGB1 expression. In contrast, AAV8-mediated overexpression of Hmgb2 enchanced CCl4-incuded liver fibrosis. Primary hepatic stellate cells (HSCs) isolated from Hmgb2-/- mice showed significantly impaired transdifferentiation and diminished activation of α-SMA, despite a modest induction of HMGB1 protein. RNA-seq analysis revealed the induction of top 45 CCl4-activated genes in multiple signaling pathways including integrin signaling and inflammation. The activation of these genes by CCl4 were abolished in Hmgb2-/- mice or in ICM-treated mice. These included C-X3-C motif chemokine receptor 1 (Cx3cr1) associated with inflammation, cyclin B (Ccnb) associated with cell cycle, DNA topoisomerase 2-alpha (Top2a) associated with intracellular component, and fibrillin (Fbn) and fibromodulin (Fmod) associated with extracellular matrix. We conclude that HMGB2 is indispensable for stellate cell activation. Therefore, HMGB2 may serve as a potential therapeutic target to prevent HSC activation during chronic liver injury. The blood HMGB2 level may also serve as a potential diagnostic marker to detect early stage of liver fibrosis and cirrhosis in humans.

Antifibrotic Efficacy of a New Phytocomposition of Essential Phospholipids with Glycyrrhizic Acid, Ecdysterone, Lycopene and Proanthacyanidin in Experimental Severe Chronic Hepatitis Compared with Phosphogliv

Since the prevalence of acute and especially severe chronic liver diseases of various etiologies increases from year to year, this pathology is recognized as the main burden on health worldwide. Currently, it has been established that the use of drugs based on essential phospholipids and plant origin with antioxidant and hepatoprotective activity is very effective in the prevention and treatment of liver diseases. In this regard, we studied under experimental conditions the effect of phytocomposition of soy lecithin, glycyrrhizic acid, lycopene and ecdysterone (conditional name hepalipin) and proanthacyanidin (conditional name yantacin) isolated from the plant Alhagi pseudalhagi on cytolytic-cholestatic liver damage, as well as on the development of fibrosis. In this study, we evaluated the hepatoprotective and antifibrotic effects of a new combination called Hepatocin obtained in a 1:1 ratio (100 mg/kg of Hepalipin and 100 mg/kg of Yantacin). All the studies conducted were conducted on adult nonlinear rats, while the experimental animals were divided into a control group infected with heliotrin, a substance with hepatotoxic action, a group receiving hepatocin, and an intact group that was not infected with heliotrin at the same time. In the conducted studies, hepatocin significantly inhibited the development of cytolytic-cholestatic liver damage, helped to maintain the functions of the liver synthesizing protein and glycogen, and when administered to experimental animals against the background of a chronic disease developing as a result of damage by heliotrin, it has an antioxidant effect. In addition, the use of hepatocin for more than two months in chronic liver damage or early stage fibrosis led to the restoration of serum enzymes of experimental animals, as well as regulators of cytochrome P450 and b5 fibrogenesis in liver tissues, PCNA, PDGF-BB to levels almost close to the initial (intact) values. Thus, in experimental conditions of chronic severe hepatitis or early stage fibrosis, hepatocin showed statistically significant advantages over Phosphogliv in terms of the intensity of hepatoprotective or antifibrotic action.

Optimization of early diagnosis of fibrosis in patients with non-alcoholic fatty liver disease with obesity and overweight

Objective — to optimize of early diagnosis of fibrosis in patients with NAFLD with obesity and overweight based on ROC analysis of non‑invasive laboratory and instrumental diagnostic markers. Materials and methods. The study included 120 patients with NAFLD (main group) with 1 — 3 degrees of obesity or overweight (BMI ≥25.0 — 39.9 kg/m2) and 20 practically healthy volunteers (control group) 23.50 [21.35; 24.78] kg/m2. Patients in the main group were divided into two subgroups depending on BMI. The subgroup I included 85 patients with NAFLD with concomitant obesity, BMI 36.50 [32.00; 40.60] kg/m2, and the subgroup II included 35 patients with NAFLD and overweight 28.00 [27.10; 29.35] kg/m2. Determination of the degree of fibrosis according to the METAVIR scale by measuring the average stiffness of the liver parenchyma, in the mode of shear wave elastography (ultrasound scanning system Soneus P7). The composition of the gut microbiota at the level of the main phylotypes was studied by identifying total bacterial DNA and DNA of Bacteroidetes, Firmicutes, as well as their ratio (F/B ratio), by quantitative real‑time polymerase chain reaction using universal primers for the 16S rRNA gene and taxon‑specific primers. Statistical processing was performed using Statistica 13.1. The data are presented in the form of Me [LQ; UQ], where Me is the median, LQ and UQ are the lower and upper quartiles, respectively. Results. Determination of the characteristic curves obtained as a result of the analysis allowed us to identify the most informationally significant additional diagnostic indicators: CRP, TNF‑α, microRNA‑122, microRNA‑34a (RU), the percentage of visceral fat, and F/B ratio. The analysis of ROC curves allowed us to determine the cut‑off value for each of the additional diagnostic criteria: cut‑off value for serum CRP concentration — 4.5 mg/l (AUC=0.99, sensitivity 1.00, specificity 0.030; p &lt;0.05); for the serum concentration of TNF‑α — 5.5 pg/mL (AUC=0.97, sensitivity 0.98, specificity 0.025; p &lt;0.05). The cut‑off value for the serum level of ­microRNA‑122 — 12.50 RU (AUC=0.99, sensitivity 0.98, specificity 0.048; p &lt;0.05); for the serum level of microRNA‑34a — 5.50 RU (AUC=0.98, sensitivity 0.98, specificity 0.106; p &lt;0.05). The cut‑off value for the proportion of VF — 8.5% (AUC=0.99, sensitivity 1.00, specificity 0.175; p &lt;0.05); for F/B ratio — 1.51 (AUC=0.95, sensitivity 0.83, specificity 0.021; p &lt;0.05). The analysis of the studied parameters allowed us to develop an algorithm for early comprehensive diagnosis of fibrosis in patients with NAFLD with obesity and overweight. The algorithm we propose consists of the following 4 stages. Stage I includes assessment of hepatopathy and interviewing the patient to determine complaints and anamnesis to exclude etiologic factors of secondary fatty liver. Stage II includes the determination of cytolysis markers, namely ALT and GGT. Stage III includes the determination of metabolic and proinflammatory parameters: HOMA index; TC and LDL cholesterol; CRP and TNF‑α. Stage IV includes the assessment of genetic and biological factors: determination of the main phylotypes of the gut microbiota and measurement of serum concentrations of microRNA‑122 and microRNA‑34a. The algorithm allows screening patients with NAFLD with obesity and overweight and identifying patients with early stages of fibrosis through a phased examination. Conclusions. The generalization of traditional diagnostic algorithms with the results of our search for additional diagnostic criteria made it possible to create a 4‑stage algorithm for early comprehensive diagnosis of fibrosis in patients with NAFLD with concomitant obesity or overweight. The use of the developed algorithm allows stratifying patients with NAFLD with obesity and overweight by the presence of fibrosis in the early stages, including by level of care, and timely prescribing preventive and therapeutic measures to reduce the risk of development, improve prognosis and prevent progression.

Prediction of Early-stage Liver Fibrosis Using FDG-PET/CT.

18F-fluorodeoxyglucose (FDG) is known to accumulate in the liver. We investigated whether accumulation of FDG was correlated with the degree of liver fibrosis and the grade of necro-inflammatory activity. This retrospective study included 35 patients who underwent FDG-positron emission tomography (PET)/computed tomography (CT) before liver surgery. On fusion images of CT and PET, by placing regions of interest on the lateral, anterior and posterior segments of the liver and the aorta, the standardized uptake value (SUV) mean, and SUV normalized by lean body mass (SUL) mean of the liver were measured, and the ratio SUVmean liver/SUVmean aorta was calculated. According to the New Inuyama Classification, subjects were classified into three groups based on the grade of liver-fibrosis degree, i.e., F0, F1+F2 and F3+F4, and into three groups based on the grade of necro-inflammatory activity, i.e., A0, A1 and A2. Each of the above parameters was then compared among the groups using a Tukey test. Average SULmean liver values of the F0, F1+F2 and F3+F4 groups were 1.573±0.211, 1.845±0.220 and 1.716±0.119, respectively. The SULmean liver of the F1+F2 group was significantly higher than that of the F0 group (p=0.0296). No significant difference was observed for the other two parameters. None of the parameters exhibited significant difference among the A0, A1, and A2 groups. FDG accumulation in the liver may be increased in the early stage of liver fibrosis. SULmean liver could be used to determine the necessity for therapeutic intervention in chronic liver disease.

Performance of Albumin-Bilirubin (ALBI) score in comparison to other non-invasive markers in the staging of liver fibrosis in chronic HCV patients

BackgroundThe prognostic ability of albumin-bilirubin score (ALBI) to assess the hepatic dysfunction in patients with hepatocellular carcinoma (HCC) was previously studied. Its role in the staging of liver fibrosis post chronic hepatitis C Virus (HCV) infection needs to be investigated.Aimto assess the diagnostic value of the ALBI score compared to other non-invasive fibrosis scores in chronic HCV patients.MethodsThis cross-sectional study included consecutive chronic HCV patients from January 2015 till December 2018. Liver stiffness measurement (LSM) by transient elastography (TE) is currently one of the most validated noninvasive methods for liver fibrosis staging and is used in daily practice as a reference for fibrosis assessment. ALBI grade as well as Fibrosis-4 (FIB-4), Aspartate aminotransferase to platelet ratio index (APRI), LOK index and Göteborg University Cirrhosis (GUCI) scores were calculated for all of the patients.ResultsA total of 781 chronic HCV patients were included. Around 54% of them had compensated cirrhosis. GUCI score was the most sensitive one to difference between early fibrosis stages, F0 vs. F1. LOK index and ALBI score did not differ significantly between F1 and F2 stages unlike the other study markers. ROC curves revealed good diagnostic capability of FIB-4 (AUROC: 0.85, 0.84), APRI (AUROC: 0.83, 0.83) and GUCI score (AUROC: 0.83, 0.83) for detecting advanced fibrosis and cirrhosis, respectively. ALBI score had a moderate diagnostic role for diagnosing advanced fibrosis and cirrhosis, AUROC of 0.73 and 0.74 respectively. At a cutoff value of -2.95, the sensitivity of ALBI score approached 79%, the specificity was 53% for advanced fibrosis.ConclusionALBI score has a moderate diagnostic power score in the diagnosis of HCV-associated advanced liver fibrosis and cirrhosis; however, FIB-4, APRI and GUCI scores outperformed the ALBI score.

Diets with Higher Anti-inflammatory Potential Associated with Lower Risk of Development of Nonalcoholic Fatty Liver Disease and Early-Stage of Fibrosis Among US Adults.

There is limited evidence of how dietary inflammatory potential influences nonalcoholic fatty liver disease (NAFLD) progression. Our study aims to evaluate the association of dietary inflammatory index (DII) with liver fibrosis, a hallmark feature of NAFLD, among US adults. Cross-sectional data consisting of 5,506 participants in the National Health and Nutrition Examination Survey from 2011 to 2018 were used. Energy adjusted-DII (E-DII) scores were calculated using 2days of 24-h dietary recall data. We used a partial proportional odds model to determine risk at each stage of fibrosis according to the E-DII score. The weighted prevalence of NAFLD (assessed by US fatty liver index) was 34.5%, with 23.2% (assessed by NAFLD Fibrosis Score) having mild fibrosis, 10.4% intermediate fibrosis, and 0.9% advanced fibrosis. When comparing the most pro-inflammatory diets to the most anti-inflammatory diets (AIDs) in the multivariable model, the marginal probability effect (MPE) of NAFLD, mild fibrosis and intermediate fibrosis increased by 11.7% (95% CI 6.6%, 16.9%), 7.0% (95% CI 3.5%, 10.4%) and 4.0% (95% CI 0.3%, 7.5%), respectively. The MPE of advanced fibrosis was not significant (MPE = 0.7%; 95% CI -1.1%, 2.8%). Similar associations were observed when applying Fibrosis-4 and transient elastography as fibrosis diagnostic measurements. An AID was associated with lower risk of development of NAFLD and early-stage of fibrosis among US adults. But the associations became attenuated and dissipated as the fibrogenesis became severe. Further studies are needed to re-confirm our observations.

Biased Quantification of Rat Liver Fibrosis-Meta-Analysis with Practical Recommendations and Clinical Implications.

For liver fibrosis assessment, the liver biopsy is usually stained with Masson's trichrome (MT) or picrosirius red (PSR) to quantify liver connective tissue (LCT) for fibrosis scoring. However, several concerns of such semiquantitative assessments have been raised, and when searching for data on the amount of LCT in healthy rats, the results vastly differ. Regarding the ongoing reproducibility crisis in science, it is necessary to inspect the results and methods, and to design an unbiased and reproducible method of LCT assessment. We searched the Medline database using search terms related to liver fibrosis, LCT and collagen, rat strains, and staining methods. Our search identified 74 eligible rat groups in 57 studies. We found up to 170-fold differences in the amount of LCT among healthy Wistar and Sprague-Dawley rats, with significant differences even within individual studies. Biased sampling and quantification probably caused the observed differences. In addition, we also found incorrect handling of liver fibrosis scoring. Assessment of LCT using stereological sampling methods (such as systematic uniform sampling) would provide us with unbiased data. Such data could eventually be used not only for the objective assessment of liver fibrosis but also for validation of noninvasive methods of the assessment of early stages of liver fibrosis.

Synergistic Modulation of a Tunable Microenvironment to Fabricate a Liver Fibrosis Chip for Drug Testing.

Liver fibrosis is a progressive physiological change that occurs after liver injury and seriously endangers human health. The lack of reliable and physiologically relevant pathological models of liver fibrosis leads to a longer drug development period and sizeable economic investment. The fabrication of a biomimetic liver-on-a-chip is significant for liver disease treatment and drug development. Here, a sandwich chip with a microwell array structure in its bottom layer was fabricated to simulate the Disse space structure of hepatic sinusoids in vitro. By synergistic modulation of the cross-linking degree of gelatin-methacryloyl (GelMA) hydrogels and the induction of transforming growth factor-beta (TGF-β), the early and late stages of liver fibrosis were designed in the chip. Owing to its three-dimensional-mixed-culture strategy, it was possible to construct a liver sinusoid model in vitro to allow for faithful physiological emulation. The model was further subjected to drug treatment, and it presented a significant difference in treatment response in early and late fibrosis progression. Our system provides a unique method for emulating liver function through a vitro liver fibrosis-on-a-chip, potentially paving the way for investigating human liver fibrosis and related drug development.

Fibroblast activation in response to TGFβ1 is modulated by co-culture with endothelial cells in a vascular organ-on-chip platform.

Background: Tissue fibrosis is a major healthcare burden that affects various organs in the body for which no effective treatments exist. An underlying, emerging theme across organs and tissue types at early stages of fibrosis is the activation of pericytes and/or fibroblasts in the perivascular space. In hepatic tissue, it is well known that liver sinusoidal endothelial cells (EC) help maintain the quiescence of stellate cells, but whether this phenomenon holds true for other endothelial and perivascular cell types is not well studied. Methods: The goal of this work was to develop an organ-on-chip microvascular model to study the effect of EC co-culture on the activation of perivascular cells perturbed by the pro-fibrotic factor TGFβ1. A high-throughput microfluidic platform, PREDICT96, that was capable of imparting physiologically relevant fluid shear stress on the cultured endothelium was utilized. Results: We first studied the activation response of several perivascular cell types and selected a cell source, human dermal fibroblasts, that exhibited medium-level activation in response to TGFβ1. We also demonstrated that the PREDICT96 high flow pump triggered changes in select shear-responsive factors in human EC. We then found that the activation response of fibroblasts was significantly blunted in co-culture with EC compared to fibroblast mono-cultures. Subsequent studies with conditioned media demonstrated that EC-secreted factors play at least a partial role in suppressing the activation response. A Luminex panel and single cell RNA-sequencing study provided additional insight into potential EC-derived factors that could influence fibroblast activation. Conclusion: Overall, our findings showed that EC can reduce myofibroblast activation of perivascular cells in response to TGFβ1. Further exploration of EC-derived factors as potential therapeutic targets in fibrosis is warranted.