Reversal Of Fibrosis Research Articles

Hydroxycamptothecin and Substratum Stiffness Synergistically Regulate Fibrosis of Human Corneal Fibroblasts.

Corneal fibrosis is a common outcome of inappropriate repair associated with trauma or ocular infection. Altered biomechanical properties with increased corneal stiffness is a feature of fibrosis that cause corneal opacities, resulting in severe visual impairment and even blindness. The present study aims to determine the effect of hydroxycamptothecin (HCPT) and matrix stiffness on transforming growth factor-β1 (TGF-β1)-induced fibrotic processes in human corneal fibroblasts (HTK cells). HTK cells were cultured on substrates with different stiffnesses ("soft", ∼261 kPa; "stiff", ∼2.5 × 103 kPa) and on tissue culture plastic (TCP, ∼106 kPa) and simultaneously treated with or without 1 μg/mL HCPT and 10 ng/mL TGF-β1. We found that HCPT induced decreased cell viability and antiproliferative effects on HTK cells. TGF-β1-induced expression of fibrosis-related genes (FN1, ACTA2) was reduced if the cells were simultaneously treated with HCPT. Substrate stiffness did not affect the expression of fibrosis-related genes. The TGF-β1 induced expression of FN1 on both soft and stiff substrates was reduced if cells were simultaneously treated with HCPT. However, this trend was not seen for ACTA2, i.e., the TGF-β1 induced expression of ACTA2 was not reduced by simultaneous treatment of HCPT in either soft or stiff substrate. Instead, HCPT treatment in the presence of TGF-β1 resulted in increased gene expression of keratocyte phenotype makers (LUM, KERA, AQP1, CHTS6) on both substrate stiffnesses. In addition, the protein expression of keratocyte phenotype makers LUM and ALDH3 was increased in HTK cells simultaneously treated with TGF-β1 and HCPT on stiff substrate as compared to control, i.e., without HCPT. In conclusion, we found that HCPT can reduce TGF-β1-induced fibrosis and promote the keratocyte phenotype in a substrate stiffness dependent manner. Thus, HCPT stimulation might be an approach to stimulate keratocytes in the appropriate healing stage to avoid or reverse fibrosis and achieve more optimal corneal wound healing.

In This Issue

HomeCirculation ResearchVol. 132, No. 1In This Issue Free AccessIn BriefPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessIn BriefPDF/EPUBIn This Issue Ruth Williams Ruth WilliamsRuth Williams Search for more papers by this author Originally published5 Jan 2023https://doi.org/10.1161/RES.0000000000000587Circulation Research. 2023;132:2Maintenance of Enteral ACE2 Prevents Diabetic Retinopathy in Type 1 Diabetes (p e1)Download figureDownload PowerPointProbiotic delivery of ACE2 improves gut integrity and retinopathy in diabetic mice, show Prasad et al.Type 1 diabetes has a complex etiology and pathology that are not entirely understood. In addition to the destruction of insulin-producing cells, a recently discovered feature of the disease (in both model rodents and humans) is that levels of angiotensin-converting enzyme 2 (ACE2)—a component of the renin-angiotensin system controlling hemodynamics—can be unusually low in certain tissues. Moreover, genetic deficiency of ACE2 in rodents, exacerbates aspects of diabetes such as gut permeability, systemic inflammation and diabetic retinopathy, while boosting ACE2 levels has been shown to ameliorate diabetic retinopathy in mice. Prasad and colleagues now show that ACE2 treatment also improves gut integrity and systemic inflammation as well as retinopathy, and that certain improvements can even be seen once diabetes has long been established. Six months after the onset of diabetes in model mice, oral doses of Lactobacillus paracasei engineered to express humanized ACE2 led to a reversal of the animal’s gut barrier dysfunction and retinopathy, the team showed. Humans with diabetic retinopathy also displayed evidence of increased gut permeability and low ACE2 levels, suggesting they may benefit from a similar probiotic treatment developed for clinical use.Epsin Nanotherapy Regulates Cholesterol Transport to Fortify Atheroma Regression (p e22)Download figureDownload PowerPointSuppressing epsin activity could be a new way to treat atherosclerosis, suggest Cui et al.Epsins are a family of plasma membrane proteins that drive endocytosis. They are expressed at varying levels throughout the tissues of the body, and recent research shows they are unusually abundant on macrophages within atherosclerotic lesions. Furthermore, in mice, macrophage-specific epsin loss results in a reduction in foam cell formation and plaque development. Cui and colleagues now show this effect on foam cells is because epsins normally promote the internalization of lipids into macrophages through their endocytic activity. But that’s not all. The proteins also impede cholesterol efflux from macrophages further exacerbating lipid retention, the team found. It turns out, epsins drive the endocytosis and degradation of a cholesterol efflux factor called ABCG1. Importantly, these proatherogenic activities of epsins could be stopped. Using macrophage-targeting nanoparticles carrying epsin-specific silencing RNAs, the team could suppress production of the protein in cultured macrophages, and could reduce the size and number of plaques in atherosclerosis-prone mice (via injection of the nanoparticles). Together the results suggest blocking epsins, via nanotherapy or other means, could be a therapeutic approach to stopping or slowing atherosclerosis progression.Inhibition of Eicosanoid Degradation Mitigates Fibrosis of the Heart (p 10)Download figureDownload PowerPointBlocking 15-PGDH quells fibroblast activation to prevent cardiac fibrosis in mice, report Rubino et al.After a myocardial infarction, the initial scar that forms is essential for protecting the muscle from further damage and rupture. But, if the fibrosis is extensive or prolonged, it can limit muscle function and ultimately lead to heart failure and death. Finding treatments that prevent or reverse fibrosis is thus a desirable clinical goal. To identify inhibitors of profibrotic activity, Rubino and colleagues screened a library of 546 known non-toxic compounds for their ability to block growth factor-induced activation of rat fibroblast cells. Of the nine compounds with such activity, the team focused on one—SW033291—that had previously been shown to prevent lung fibrosis in mice. This compound inhibits the enzyme 15-PGDH, which degrades signaling molecules known as eicosanoids, which are involved in, among other things, tissue repair. Further experiments showed the compound could also switch off the activation state of human cardiac fibroblasts isolated from failing hearts. The team went on to show that infusing SW033291 into mice could ameliorate experimentally induced cardiac fibrosis and diastolic dysfunction. The findings thus identify 15-PDGH as a profibrotic factor that could serve as a therapeutic target for mitigating heart fibrosis and failure. Previous Back to top Next FiguresReferencesRelatedDetails January 6, 2023Vol 132, Issue 1 Advertisement Article InformationMetrics © 2022 American Heart Association, Inc.https://doi.org/10.1161/RES.0000000000000587 Originally publishedJanuary 5, 2023 PDF download Advertisement

Quantitative proteomics analysis revealed the potential role of lncRNA Ftx in cardiomyocytes

ObjectiveThis study aims to decode the proteomic signature of cardiomyocytes in response to lncRNA Ftx knockdown and overexpression via proteomic analysis, and to study the biological role of lncRNA Ftx in cardiomyocytes. MethodsThe expression level of the lncRNA Ftx in cardiomyocytes cultured in vitro was intervened, and the changes in protein levels in cardiomyocytes were quantitatively detected by liquid chromatography-mass spectrometry. The key molecules and pathways of the lncRNA-Ftx response were further examined by GO, KEGG, and protein interaction analysis.ResultsA total of 2828 proteins are quantified. With a 1.5-fold change threshold, 32 upregulated proteins and 49 downregulated proteins are identified in the lncRNA Ftx overexpression group, while 67 up-regulated proteins and 54 down-regulated proteins are identified in the lncRNA Ftx knockdown group. Functional clustering analysis of differential genes revealed that the lncRNA Ftx is involved in regulating cardiomyocyte apoptosis and ferroptosis and improving cellular energy metabolism. In addition, Hub genes such as ITGB1, HMGA2, STAT3, GSS, and LPCAT3 are regulated downstream by lncRNA Ftx.ConclusionThis study demonstrates that lncRNA Ftx plays a vital role in cardiomyocytes and may be involved in the occurrence and development of various myocardial diseases. It provides a potential target for clinical protection of the myocardium and reversal of myocardial fibrosis.

Telocyte-Derived Exosomes Provide an Important Source of Wnts That Inhibits Fibrosis and Supports Regeneration and Repair of Endometrium

Intrauterine adhesions (IUAs) often occurred after common obstetrical and gynecological procedures or infections in women of reproductive age. It was characterized by the formation of endometrial fibrosis and prevention of endometrial regeneration, usually with devastating fertility consequences and poor treatment outcomes so far. Telocytes (TCs), as a novel interstitial cell type, present in female uterus with in vitro therapeutic potential in decidualization-defective gynecologic diseases. This study aims to further investigate the role of TC-derived Wnt ligands carried by exosomes (Exo) in reversal of fibrosis and enhancement of regeneration repair in endometrium. IUA cellular and animal models were established from endometrial stromal cells (ESCs) and mice, followed with treatment of TC-conditioned medium (TCM) or TC-derived Exo. In cellular model, fibrosis markers (collagen type 1 alpha 1 [COL1A1], fibronectin [FN], and α-smooth muscle actin [α-SMA]), angiogenesis (vascular endothelial growth factor [VEGF]), and pathway protein (β-catenin) were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR), Western blotting (WB), and immunofluorescence. Results showed that, TCs (either TCM or TC-derived Exo) provide a source of Wnts that inhibit cellular fibrosis, as evidenced by significantly elevated VEGF and β-catenin with decreased fibrotic markers, whereas TCs lost salvage on fibrosis after being blocked with Wnt/β-catenin inhibitors (XAV939 or ETC-159). Further in mouse model, regeneration repair (endometrial thickness, number of glands, and fibrosis area ratio), fibrosis markers (fibronectin [FN]), mesenchymal–epithelial transition (MET) (E-cadherin, N-cadherin), and angiogenesis (VEGF, microvessel density [MVD]) were studied by hematoxylin–eosin (HE), Masson staining, and immunohistochemistry. Results demonstrated that TC-Exo treatment effectively promotes regeneration repair of endometrium by relieving fibrosis, enhancing MET, and angiogenesis. These results confirmed new evidence for therapeutic perspective of TC-derived Exo in IUAs.

Direct Reprogramming Improves Cardiac Function and Reverses Fibrosis in Chronic Myocardial Infarction.

Because adult cardiomyocytes have little regenerative capacity, resident cardiac fibroblasts (CFs) synthesize extracellular matrix after myocardial infarction (MI) to form fibrosis, leading to cardiac dysfunction and heart failure. Therapies that can regenerate the myocardium and reverse fibrosis in chronic MI are lacking. The overexpression of cardiac transcription factors, including Mef2c/Gata4/Tbx5/Hand2 (MGTH), can directly reprogram CFs into induced cardiomyocytes (iCMs) and improve cardiac function under acute MI. However, the ability of in vivo cardiac reprogramming to repair chronic MI with established scars is undetermined. We generated a novel Tcf21iCre/reporter/MGTH2A transgenic mouse system in which tamoxifen treatment could induce both MGTH and reporter expression in the resident CFs for cardiac reprogramming and fibroblast lineage tracing. We first tested the efficacy of this transgenic system in vitro and in vivo for acute MI. Next, we analyzed in vivo cardiac reprogramming and fusion events under chronic MI using Tcf21iCre/Tomato/MGTH2A and Tcf21iCre/mTmG/MGTH2A mice, respectively. Microarray and single-cell RNA sequencing were performed to determine the mechanism of cardiac repair by in vivo reprogramming. We confirmed the efficacy of transgenic in vitro and in vivo cardiac reprogramming for acute MI. In chronic MI, in vivo cardiac reprogramming converted ≈2% of resident CFs into iCMs, in which a majority of iCMs were generated by means of bona fide cardiac reprogramming rather than by fusion with cardiomyocytes. Cardiac reprogramming significantly improved myocardial contraction and reduced fibrosis in chronic MI. Microarray analyses revealed that the overexpression of MGTH activated cardiac program and concomitantly suppressed fibroblast and inflammatory signatures in chronic MI. Single-cell RNA sequencing demonstrated that resident CFs consisted of 7 subclusters, in which the profibrotic CF population increased under chronic MI. Cardiac reprogramming suppressed fibroblastic gene expression in chronic MI by means of conversion of profibrotic CFs to a quiescent antifibrotic state. MGTH overexpression induced antifibrotic effects partly by suppression of Meox1, a central regulator of fibroblast activation. These results demonstrate that cardiac reprogramming could repair chronic MI by means of myocardial regeneration and reduction of fibrosis. These findings present opportunities for the development of new therapies for chronic MI and heart failure.

Fibrosis Changes Following Viral Clearance and its Relation to HCC

Globally, about 290 000 people died from hepatitis C (HCV) in 2019, mostly from cirrhosis and hepatocellular carcinoma (HCC). Less than a decade, direct-acting antivirals (DAAs) replaced interferon-based therapies in treatment protocols and sustained virologic response (SVR) rates exceeded 95%. However, it is not clear yet whether HCV eradication by DAAs can reverse fibrosis or cirrhosis substantially, reduce the risk of developing HCC in the long term. In this review was aimed to summarizes the evidence for reversibility or progression of fibrosis following HCV clearance by DAAs, subsequently its impact on developing de nova HCC. For this purpose, cohorts in which patients achieved SVR, have paired liver stiffness measurements obtained by transient elastography were reviewed and results discussed.

Suppression of pullulanase-induced cytotoxic T cell response with a dual promoter in GSD IIIa mice.

Glycogen debranching enzyme deficiency in glycogen storage disease type III (GSD III) results in excessive glycogen accumulation in multiple tissues, primarily the liver, heart, and skeletal muscle. We recently reported that an adeno-associated virus vector expressing a bacterial debranching enzyme (pullulanase) driven by the ubiquitous CMV enhancer/chicken β-actin (CB) promoter cleared glycogen in major affected tissues of infant GSD IIIa mice. In this study, we developed a potentially novel dual promoter consisting of a liver-specific promoter (LSP) and the CB promoter for gene therapy in adult GSD IIIa mice. Ten-week treatment with an adeno-associated virus vector containing the LSP-CB dual promoter in adult GSD IIIa mice significantly increased pullulanase expression and reduced glycogen contents in the liver, heart, and skeletal muscle, accompanied by the reversal of liver fibrosis, improved muscle function, and a significant decrease in plasma biomarkers alanine aminotransferase, aspartate aminotransferase, and creatine kinase. Compared with the CB promoter, the dual promoter effectively decreased pullulanase-induced cytotoxic T lymphocyte responses and enabled persistent therapeutic gene expression in adult GSD IIIa mice. Future studies are needed to determine the long-term durability of dual promoter-mediated expression of pullulanase in adult GSD IIIa mice and in large animal models.

Remodeling of imbalanced extracellular matrix homeostasis for reversal of pancreatic fibrosis

Pancreatic fibrosis is mainly manifested by imbalance in extracellular matrix (ECM) homeostasis due to excessive deposition of collagen in pancreas by activated pancreatic stellate cells (PSCs). Recently, some drugs have exhibited therapeutic potentials for the treatment of pancreatic fibrosis; however, currently, no effective clinical strategy is available to remodel imbalanced ECM homeostasis because of inferior targeting abilities of drugs and collagen barriers that hinder the efficient delivery of drugs. Herein, we design and prepare collagen-binding peptide (CBP) and collagenase I co-decorated dual drug-loaded lipid nanoparticles (named AT-CC) for pancreatic fibrosis therapy. Specifically, AT-CC can target fibrotic pancreas via the CBP and degrade excess collagen by the grafted collagenase I, thereby effectively delivering all-trans-retinoic acid (ATRA) and ammonium tetrathiomolybdate (TM) into pancreas. The released ATRA can reduce collagen overproduction by inhibiting the activation of PSCs. Moreover, the released TM can restrain lysyloxidase activation, consequently reducing collagen cross-linking. The combination of ATRA and TM represses collagen synthesis and reduces collagen cross linkages to restore ECM homeostasis. The results of this research suggest that AT-CC is a safe and efficient collagen-targeted degradation drug-delivery system for reversing pancreatic fibrosis. Furthermore, the strategy proposed herein will offer an innovative platform for the treatment of chronic pancreatitis.

EARLY DETECTION AND VALIDATION OF PREVENTIVE MEASURES OF ORAL SUBMUCOSAL FIBROSIS WITH SPECIAL REFERENCE TO SARVASARA MUKHAROGA: A LITERARY REVIEW

The oral cavity is considered a reflection of the body's internal systems. As a result, significant efforts must be done to keep it healthy and free of chronic illnesses that may result in unfavourable outcomes. Oral Submucous Fibrosis (OSMF) is a chronic debilitating condition of the oral cavity that involves the formation of constricting bands of collagen fibre in the oral mucosa over time, resulting in restricted movement of the mouth and tongue and eventually hindered or problematic speech and difficulty swallowing food. Many pharmacological and surgical techniques have been used in modern terms, but the outcomes have been unsatisfactory due to recurrence and dire consequences. When the symptomatology of OSMF is compared to Ayurvedic classics, it can be classified as Vata Pitta Pradhana Sarvasara Mukharoga. An attempt was made here to explore preventive measures for OSMF using Ayurvedic methods. This study will discuss treatment at both the local and systemic levels. Local modalities such as vrana shodhana and, vrana ropana pratisarana (rubbing) & kavala (gargling) halt disease progression, and these local therapies prevent and reverse fibrosis while also improving motor functions of the mouth. Systemic therapy restores balance and improves the oral mucosa's vitality. This study emphasizes how the treatment approach is superior to all other surgical and nonsurgical treatments. Early withdrawal of destructive behaviours and adequate therapy with ayurvedic medications aid in managing OSMF.

Activated Hepatic Stellate Cells in Hepatocellular Carcinoma: Their Role as a Potential Target for Future Therapies

Hepatocellular carcinoma (HCC) is a global healthcare challenge, which affects more than 815,000 new cases every year. Activated hepatic stellate cells (aHSCs) remain the principal cells that drive HCC onset and growth. aHSCs suppress the anti-tumor immune response through interaction with different immune cells. They also increase the deposition of the extracellular matrix proteins, challenging the reversion of fibrosis and increasing HCC growth and metastasis. Therapy for HCC was reported to activate HSCs, which could explain the low efficacy of current treatments. Conversely, recent studies aimed at the deactivation of HSCs show that they have been able to inhibit HCC growth. In this review article, we discuss the role of aHSCs in HCC pathophysiology and therapy. Finally, we provide suggestions for the experimental implementation of HSCs in HCC therapies.

Combination of esomeprazole and pirfenidone enhances antifibrotic efficacy in vitro and in a mouse model of TGFβ-induced lung fibrosis

Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal lung disease of unknown etiology. Currently, pirfenidone and nintedanib are the only FDA-approved drugs for the treatment of IPF and are now the standard of care. This is a significant step in slowing down the progression of the disease. However, the drugs are unable to stop or reverse established fibrosis. Several retrospective clinical studies indicate that proton pump inhibitors (PPIs; FDA-approved to treat gastroesophageal reflux) are associated with favorable outcomes in patients with IPF, and emerging preclinical studies report that PPIs possess antifibrotic activity. In this study, we evaluated the antifibrotic efficacy of the PPI esomeprazole when combined with pirfenidone in vitro and in vivo. In cell culture studies of IPF lung fibroblasts, we assessed the effect of the combination on several fibrosis-related biological processes including TGFβ-induced cell proliferation, cell migration, cell contraction, and collagen production. In an in vivo study, we used mouse model of TGFβ-induced lung fibrosis to evaluate the antifibrotic efficacy of esomeprazole/pirfenidone combination. We also performed computational studies to understand the molecular mechanisms by which esomeprazole and/or pirfenidone regulate lung fibrosis. We found that esomeprazole significantly enhanced the anti-proliferative effect of pirfenidone and favorably modulated TGFβ-induced cell migration and contraction of collagen gels. We also found that the combination significantly suppressed collagen production in response to TGFβ in comparison to pirfenidone monotherapy. In addition, our animal study demonstrated that the combination therapy effectively inhibited the differentiation of lung fibroblasts into alpha smooth muscle actin (αSMA)-expressing myofibroblasts to attenuate the progression of lung fibrosis. Finally, our bioinformatics study of cells treated with esomeprazole or pirfenidone revealed that the drugs target several extracellular matrix (ECM) related pathways with esomeprazole preferentially targeting collagen family members while pirfenidone targets the keratins. In conclusion, our cell biological, computational, and in vivo studies show that the PPI esomeprazole enhances the antifibrotic efficacy of pirfenidone through complementary molecular mechanisms. This data supports the initiation of prospective clinical studies aimed at repurposing PPIs for the treatment of IPF and other fibrotic lung diseases where pirfenidone is prescribed.

Niclosamide-loaded nanoparticles (Ncl-NPs) reverse pulmonary fibrosis in vivo and in vitro

IntroductionIdiopathic pulmonary fibrosis (IPF), a life-threatening interstitial lung disease, is characterized by excessive activation and proliferation of fibroblasts and epithelial-mesenchymal transition (EMT) of alveolar epithelial cells (AEC) accompanied by a large amount of extracellular matrix aggregation. There are no therapies to reverse pulmonary fibrosis, and nintedanib and pirfenidone could only slow down the decline of lung function of IPF patients and delay their survival time. Niclosamide (Ncl) is an antihelminthic drug approved by FDA, which has been reported to have pleiotropic pharmacological activities in recent years, but it’s almost complete insolubility in water limits its clinical application. ObjectivesTo improve the water solubility of Ncl, explore its ability to reverse BLM-induced pulmonary fibrosis and its specific mechanism of action. MethodsThe Niclosamide-loaded nanoparticles (Ncl-NPs) were formed by emulsification solvent evaporation method. A mouse model induced by bleomycin (BLM) was established to evaluate its effects and mechanisms of inhibiting and reversing fibrosis in vivo. The cell models treated by transforming growth factor-β1 (TGF-β1) were used to examine the mechanism of Ncl-NPs inhibiting fibrosis in vitro. Flow cytometry, IHC, IL-4-induced macrophage model and co-culture system were used to assess the effect of Ncl-NPs on M2 polarization of macrophages. ResultsThe Ncl-NPs improved the poor water solubility of Ncl. The lower dose of Ncl-NPs (2.5 mg/kg) showed the same effect of reversing established pulmonary fibrosis as free Ncl (5 mg/kg). Mechanistic studies revealed that Ncl-NPs blocked TGF-β/Smad and signaling transducer and activator of transcription 3 (Stat3) signaling pathways and inhibited the M2 polarization of macrophages. Additionally, H&E staining of the tissues initially showed the safety of Ncl-NPs. ConclusionThese results indicate Ncl-NPs may serve as a new idea for the treatment of pulmonary fibrosis.

ODP005 Molecular Characterization of Early Adipose Tissue Remodeling and Fibrosis in a Murine Model of Diet-Induced Obesity.

Abstract One of the many pathophysiological changes associated with obesity is the build up of structural and physiochemical extracellular matrix components in adipose depots, known as adipose tissue fibrosis. Like liver fibrosis, adipose tissue fibrosis occurs as a response to chronic injury and inflammation and can perpetuate inflammation and impede proper tissue functioning. Key adipose tissue fibrosis markers have been identified in obese adipose tissue, including collagen VI and its cleavage product endotrophin, however, adipose tissue fibrosis remains poorly characterized, particularly in its initiating phases. We have developed and characterized a murine model of diet-induced obesity (12 weeks of high fat feeding), insulin resistance, and early fibrotic development in both sexes. While diagnosable fibrosis was absent, as determined by picrosirius red staining, we detected elevated collagen VI, TGF-β, TNF-α, and TIMP-1 mRNA in gonadal white adipose of both sexes. TIMP-4 was decreased in males but elevated in females. Immunohistochemical analysis revealed pericellular build up of collagen VI immunoreactivity in males only, co-localized with immunoreactivity of its protease MMP-14, which is known to generate the pro-fibrotic fragment endotrophin. This work identifies key sex differences and fibrosis markers present during the initiation of adipose tissue fibrosis, earlier than diagnosable fibrosis can be detected, highlighting their potential as therapeutic targets for prevention or reversal of fibrosis in the treatment of obesity and insulin resistance. Presentation: No date and time listed

Digital pathology with artificial intelligence analyses provides greater insights into treatment-induced fibrosis regression in NASH.

Liver fibrosis is a key prognostic determinant for clinical outcomes in non-alcoholic steatohepatitis (NASH). Current scoring systems have limitations, especially in assessing fibrosis regression. Second harmonic generation/two-photon excitation fluorescence (SHG/TPEF) microscopy with artificial intelligence analyses provides standardized evaluation of NASH features, especially liver fibrosis and collagen fiber quantitation on a continuous scale. This approach was applied to gain in-depth understanding of fibrosis dynamics after treatment with tropifexor (TXR), a non-bile acid farnesoid X receptor agonist in patients participating in the FLIGHT-FXR study (NCT02855164). Unstained sections from 198 liver biopsies (paired: baseline and end-of-treatment) from 99 patients with NASH (fibrosis stage F2 or F3) who received placebo (n= 34), TXR 140μg (n= 37), or TXR 200μg (n= 28) for 48 weeks were examined. Liver fibrosis (qFibrosis®), hepatic fat (qSteatosis®), and ballooned hepatocytes (qBallooning®) were quantitated using SHG/TPEF microscopy. Changes in septa morphology, collagen fiber parameters, and zonal distribution within liver lobules were also quantitatively assessed. Digital analyses revealed treatment-associated reductions in overall liver fibrosis (qFibrosis®), unlike conventional microscopy, as well as marked regression in perisinusoidal fibrosis in patients who had either F2 or F3 fibrosis at baseline. Concomitant zonal quantitation of fibrosis and steatosis revealed that patients with greater qSteatosis reduction also have the greatest reduction in perisinusoidal fibrosis. Regressive changes in septa morphology and reduction in septa parameters were observed almost exclusively in F3 patients, who were adjudged as 'unchanged' with conventional scoring. Fibrosis regression following hepatic fat reduction occurs initially in the perisinusoidal regions, around areas of steatosis reduction. Digital pathology provides new insights into treatment-induced fibrosis regression in NASH, which are not captured by current staging systems. The degree of liver fibrosis (tissue scarring) in non-alcoholic steatohepatitis (NASH) is the main predictor of negative clinical outcomes. Accurate assessment of the quantity and architecture of liver fibrosis is fundamental for patient enrolment in NASH clinical trials and for determining treatment efficacy. Using digital microscopy with artificial intelligence analyses, the present study demonstrates that this novel approach has greater sensitivity in demonstrating treatment-induced reversal of fibrosis in the liver than current systems. Furthermore, additional details are obtained regarding the pathogenesis of NASH disease and the effects of therapy.

Fat transplantation induces dermal adipose regeneration and reverses skin fibrosis through dedifferentiation and redifferentiation of adipocytes

BackgroundLocalized scleroderma causes cosmetic disfigurement, joint contractures, and other functional impairment, but no currently available medications can reverse the resulting skin lesions. Fat grafting is beneficial for reversing skin fibrosis; however, the mechanism by which adipose tissue transplantation contributes to lesion improvement has not been fully clarified. The purpose of our study was to verify the therapeutic effect of fat grafts in reversing skin fibrosis.MethodsInguinal fat pads from AdipoqCreER+;mT/mG mice, which were treated with tamoxifen, were transplanted to the skin lesion in bleomycin-treated wild-type C57 mice. Tdtomato transgenic mice-derived adipocytes, adipose-derived stem cells (ASCs), dedifferentiated adipocytes (DAs) were embedded in matrigel and transplanted beneath the skin lesion of bleomycin-treated wild-type C57 mice. A transwell co‐culture system was used to verify the effect of ASCs, adipocytes or DAs on scleroderma fibroblasts or monocytes.ResultsAdipocytes from the fat grafts could undergo dedifferentiation and redifferentiation for dermal adipose tissue re-accumulation within the skin lesion. Moreover, compared with ASCs and adipocytes, DAs show greater potency of inducing adipogenesis. ASCs and DAs showed comparable effect on inducing angiogenesis and suppressing macrophage infiltration in fibrotic skin. Co-culture assay showed that DAs and ASCs were able to reduce fibrosis-related genes in human scleroderma fibroblasts and drive M2 macrophage polarization.ConclusionOur results indicated that adipocytes would transform into a more functional and dedifferentiated state and reverse dermal fibrosis, by promoting dermal adipose tissue regeneration, improving angiogenesis, suppressing macrophage-mediated inflammation and myofibroblast accumulation.

S500 Long-Term Follow-Up of Esophageal Strictures in Eosinophilic Esophagitis Using Structured Esophagram Protocol

Introduction: Reversal of transmural fibrostenosis in eosinophilic esophagitis (EoE) is not well studied. Our aim was to determine the effect of medical therapy, dilation and initial diameter on esophageal lumen diameter using serial structured esophagrams over a period of years. Methods: Retrospective study of 78 patients who completed two EoE protocol esophagrams at an academic tertiary referral center 2003 to 2021. Maximum and minimum esophageal diameters were measured on images during rapid swallowing in the RAO recumbent position. EoE was diagnosed by consensus definition and classified as active using ≥15 eosinophils per high power field (hpf). Demographics, medical therapies, and endoscopic data were obtained by chart review. Change in esophageal diameter was analyzed with Wilcoxon signed rank test and reported as median, 25th percentile (Q1), and 75th percentile (Q3) values. Results: Median age at first esophagram was 36.2 and 60.3% were male. Medical therapies during last esophagram were PPI (39.5%), swallowed topical steroids (31.6%), diet elimination (13.2%), biologic therapies (1.3%), and clinical trials (1.3%). Eleven patients had dilation before the first esophagram and 33 between esophagrams without significant effect on results. Median years between esophagrams was 2.6 (Table). Median maximum diameter significantly increased by 1.0 mm (Q1: -1.0 mm, Q3: 3.0 mm) (P=0.034) independent of dilation (P=0.744). Median maximum diameter change per year significantly increased by 0.4 mm (Q1: -0.4 mm, Q3: 1.3 mm, P=0.010). The increase appeared most profound in patients starting in the lowest maximum diameter group (9-15 mm) with median increase of 3.0 mm while the highest starting maximum diameter group ( >21 mm) had further narrowing by 2.0 mm (Figure). There was no difference in maximum diameter change for patients on medical therapy compared to no therapy at second esophagram at 1.0 mm (Q1: -1.0 mm Q3: 3.0 mm) and 1.0 mm (Q1: 0.0 mm Q3: 2.0 mm) respectively (P=0.640); however, for patients in disease remission at second esophagram, there was a significant increase in maximum diameter per year compared to active disease at 0.8 mm (Q1: 0.0 mm Q3: 5.3 mm) and 0.0 mm (Q1: -0.4 mm Q3: 0.6 mm) respectively (P=0.019). Conclusion: Long term medical therapy leads to a small, but significant improvement in esophageal diameter in EoE. Whether this improvement is due to reversal of fibrosis or transmural inflammation is unclear.Figure 1.: Maximum Diameter Change Between Esophagram 1 and 2 based on starting maximum esophageal diameter. Table 1. - Esophagram Characteristics No Dilation (n=43) Dilation (n=34) P-Value Total (n=78) P-Value Median Years Between Esophagrams (Range) 2.7 (0.1-11.6) 2.4 (0.1-12.4) 2.6 (0.1-12.4) Median Maximum Diameter Change, mm 1.0 1.0 0.744 1.0 0.034 Q1, mm -1.0 0.0 -1.0 Q3, mm 2.5 3.0 3.0 Median Maximum Diameter Change Per Year, mm 0.3 0.4 0.961 0.4 0.010 Q1, mm -0.4 0.0 -0.4 Q3, mm 1.5 1.3 1.3 Median Minimum Diameter Change, mm 0.0 1.0 0.317 0.0 0.277 Q1, mm -2.0 -1.0 -1.5 Q3, mm 2.0 3.0 2.0 Median Minimum Diameter Change Per Year, mm 0.0 0.4 0.249 0.0 0.059 Q1, mm -0.7 -0.3 -0.5 Q3, mm 0.9 1.5 1.1 mm, millimeters; Q1, 25th percentile; Q3, 75th percentile

Liver cirrhosis in children – the role of imaging in the diagnostic pathway

Liver cirrhosis in children is a rare disease with multifactorial causes that are distinct from those in adults. Underlying reasons include cholestatic, viral, autoimmune, hereditary, metabolic and cardiac disorders. Early detection of fibrosis is important as clinical stabilization or even reversal of fibrosis can be achieved in some disorders with adequate treatment. This article focuses on the longitudinal evaluation of children with chronic liver disease with noninvasive imaging tools, which play an important role in detecting cirrhosis, defining underlying causes, grading fibrosis and monitoring patients during follow-up. Ultrasound is the primary imaging modality and it is used in a multiparametric fashion. Magnetic resonance imaging and computed tomography are usually applied second line for refined tissue characterization, clarification of nodular lesions and full delineation of abdominal vessels, including portosystemic communications.

Circulating microRNAs as Biomarkers of Hepatic Fibrosis in Schistosomiasis Japonica Patients in the Philippines.

Host-derived microRNAs (miRNAs) play important regulatory roles in schistosomiasis-induced hepatic fibrosis. This study analyzed selected serum miRNAs among Filipino schistosomiasis japonica patients with ultrasound (US)-detectable hepatic fibrosis. A prospective cohort study design with convenience sampling was employed from 2017 to 2019. The study sites were eight endemic barangays in Leyte, Philippines. Eligible chronic schistosomiasis patients with varying severities of hepatic fibrosis were enrolled in the cohort and serially examined at 6, 12, and 24 months from baseline. Baseline serum miR-146a-5p, let-7a-5p, miR-150-5p, miR-122-5p, miR-93-5p, and miR200b-3p were measured using RT-qPCR. A total of 136 chronic schistosomiasis patients were included in this prospective cohort study. Approximately, 42.6% had no fibrosis, 22.8% had mild fibrosis, and 34.6% had severe fibrosis at baseline The serum levels of the antifibrotic miR-146a (p < 0.0001), miR-150 (p = 0.0058), and let-7a (p < 0.0001) were significantly lower in patients with hepatic fibrosis while the profibrotic miR-93 (p = 0.0024) was elevated. miR-146a-5p (AUC = 0.90, 95% CI [0.84, 0.96], p < 0.0001) has the most promising potential to differentiate patients with (n = 78) versus without (n = 58) hepatic fibrosis. The baseline level of serum miR-146-5p was significantly different in patients with progressive fibrosis (n = 17) compared to those who never developed fibrosis (n = 30, p < 0.01) or those who had fibrosis reversal (n = 20, p < 0.01) after 24 months. These findings demonstrate the potential utility of serum miRNAs, particularly of miR-146a, as a supplementary tool for assessing hepatic fibrosis in chronic schistosomiasis japonica patients.

T cells and liver fibrosis

AbstractLiver fibrosis develops from the excessive deposition of extracellular matrix in the liver caused by chronic liver inflammation or various chronic injuries, and it eventually develops into liver cirrhosis. The process of liver fibrosis is closely related to the immune response, and increasing evidence reveals the role of T lymphocytes, including Th1, Th2, Th17, regulatory T cells, and mucosa‐associated invariant T cells, in liver fibrosis. These immune cells play antifibrotic or profibrotic roles during fibrosis, and the reversal of fibrosis by targeting immune cells has attracted widespread attention. Activation of hepatic stellate cells, which form the core of fibrosis, is regulated by various immune mediators, including various immune cells and their associated cytokines. Therefore, the mechanism of action elicited by each cell type must be further elucidated to provide a basis for the design of new therapeutic targets. The purpose of this review is to summarize the roles and mechanisms of T lymphocytes and their subsets in liver fibrosis and highlight the biomarkers and potential therapeutic targets associated with these cells.

Relaxin-2 as a Potential Biomarker in Cardiovascular Diseases.

The pleiotropic hormone relaxin-2 plays a pivotal role in the physiology and pathology of the cardiovascular system. Relaxin-2 exerts relevant regulatory functions in cardiovascular tissues through the specific receptor relaxin family peptide receptor 1 (RXFP1) in the regulation of cardiac metabolism; the induction of vasodilatation; the reversion of fibrosis and hypertrophy; the reduction of inflammation, oxidative stress, and apoptosis; and the stimulation of angiogenesis, with inotropic and chronotropic effects as well. Recent preclinical and clinical outcomes have encouraged the potential use of relaxin-2 (or its recombinant form, known as serelaxin) as a therapeutic strategy during cardiac injury and/or in patients suffering from different cardiovascular disarrangements, especially heart failure. Furthermore, relaxin-2 has been proposed as a promising biomarker of cardiovascular health and disease. In this review, we emphasize the relevance of the endogenous hormone relaxin-2 as a useful diagnostic biomarker in different backgrounds of cardiovascular pathology, such as heart failure, atrial fibrillation, myocardial infarction, ischemic heart disease, aortic valve disease, hypertension, and atherosclerosis, which could be relevant in daily clinical practice and could contribute to comprehending the specific role of relaxin-2 in cardiovascular diseases.