Biomedical subjects

Srinivasan Dasarathy

Publications and source records attributed to Srinivasan Dasarathy.

13 recordsLinked to original sources

PubMed2025

Integrated Multiomics Analyses of the Molecular Landscape of Sarcopenia in Alcohol-Related Liver Disease.

BACKGROUND: Skeletal muscle is a major target for ethanol-induced perturbations, leading to sarcopenia in alcohol-related liver disease (ALD). The complex interactions and pathways involved in adaptive and maladaptive responses to ethanol in skeletal muscle are not well understood. Unlike hypothesis-driven experiments, an integrated multiomics-experimental validation approach provides a comprehensive view of these interactions. METHODS: We performed multiomics analyses with experimental validation to identify novel regulatory mechanisms of sarcopenia in ALD. Studies were done in a comprehensive array of models including ethanol-treated (ET) murine and human-induced pluripotent stem cell-derived myotubes (hiPSCm), skeletal muscle from a mouse model of ALD (mALD) and human patients with alcohol-related cirrhosis and controls. We generated 13 untargeted datasets, including chromatin accessibility (assay for transposase accessible chromatin), RNA sequencing, proteomics, phosphoproteomics, acetylomics and metabolomics, and conducted integrated multiomics analyses using UpSet plots and feature extraction. Key findings were validated using immunoblots, redox measurements (NAD+/NADH ratio), imaging and senescence-associated molecular phenotype (SAMP) assays. Mechanistic studies included mitochondrial-targeted Lactobacillus brevis NADH oxidase (MitoLbNOX) to increase redox ratio and MitoTempo as a mitochondrial free radical scavenger. RESULTS: Multiomics analyses revealed enrichment in mitochondrial oxidative function, protein synthesis and senescence pathways consistent with the known effects of hypoxia-inducible factor 1α (HIF1α) during normoxia. Across preclinical and clinical models, HIF1α targets (n = 32 genes) and signalling genes (n > 100 genes) (n = 3 ATACseq, n = 65 phosphoproteomics, n = 10 acetylomics, n = 6 C2C12 proteomics, n = 106 C2C12 RNAseq, n = 64 hiPSC RNAseq, n = 30 hiPSC proteomics, n = 3 mouse proteomics, n = 25 mouse RNAseq, n = 8 human RNAseq, n = 3 human proteomics) were increased. Stabilization of HIF1α (C2C12, 6hEtOH 0.24 ± 0.09; p = 0.043; mALD 0.32 ± 0.074; p = 0.005; data shown as mean difference ± standard error mean) was accompanied by enrichment in the early transient and late change clusters, -log(p-value) = 1.5-3.8, of the HIF1α signalling pathway. Redox ratio was reduced in ET myotubes (C2C12: 15512 ± 872.1, p < 0.001) and mALD muscle, with decreased expression of electron transport chain components (CI-V, p < 0.05) and Sirt3 (C2C12: 0.067 ± 0.023, p = 0.025; mALD: 0.41 ± 0.12, p = 0.013). Acetylation of mitochondrial proteins was increased in both models (C2C12: 107364 ± 4558, p = 0.03; mALD: 40036 ± 18 987, p = 0.049). Ethanol-induced SAMP was observed across models (P16: C2C12: 0.2845 ± 0.1145, p < 0.05; hiPSCm: 0.2591, p = 0.041). MitoLbNOX treatment reversed redox imbalance, HIF1α stabilization, global acetylation and myostatin expression (p < 0.05). CONCLUSIONS: An integrated multiomics approach, combined with experimental validation, identifies HIF1α stabilization and accelerated post-mitotic senescence as novel mechanisms of sarcopenia in ALD. These findings show the complex molecular interactions leading to mitochondrial dysfunction and progressive sarcopenia in ALD.

Sarcopenia↗

PubMed2024

Membrane-bound O-acyltransferase 7 (MBOAT7) shapes lysosomal lipid homeostasis and function to control alcohol-associated liver injury.

Recent genome-wide association studies (GWAS) have identified a link between single-nucleotide polymorphisms (SNPs) near the MBOAT7 gene and advanced liver diseases. Specifically, the common MBOAT7 variant (rs641738) associated with reduced MBOAT7 expression is implicated in non-alcoholic fatty liver disease (NAFLD), alcohol-associated liver disease (ALD), and liver fibrosis. However, the precise mechanism underlying MBOAT7-driven liver disease progression remains elusive. Previously, we identified MBOAT7-driven acylation of lysophosphatidylinositol lipids as key mechanism suppressing the progression of NAFLD (Gwag et al., 2019). Here, we show that MBOAT7 loss of function promotes ALD via reorganization of lysosomal lipid homeostasis. Circulating levels of MBOAT7 metabolic products are significantly reduced in heavy drinkers compared to healthy controls. Hepatocyte- (Mboat7-HSKO), but not myeloid-specific (Mboat7-MSKO), deletion of Mboat7 exacerbates ethanol-induced liver injury. Lipidomic profiling reveals a reorganization of the hepatic lipidome in Mboat7-HSKO mice, characterized by increased endosomal/lysosomal lipids. Ethanol-exposed Mboat7-HSKO mice exhibit dysregulated autophagic flux and lysosomal biogenesis, associated with impaired transcription factor EB-mediated lysosomal biogenesis and autophagosome accumulation. This study provides mechanistic insights into how MBOAT7 influences ALD progression through dysregulation of lysosomal biogenesis and autophagic flux, highlighting hepatocyte-specific MBOAT7 loss as a key driver of ethanol-induced liver injury.

Animals↗

PubMed2024

Proteomics identifies complement protein signatures in patients with alcohol-associated hepatitis.

Diagnostic challenges continue to impede development of effective therapies for successful management of alcohol-associated hepatitis (AH), creating an unmet need to identify noninvasive biomarkers for AH. In murine models, complement contributes to ethanol-induced liver injury. Therefore, we hypothesized that complement proteins could be rational diagnostic/prognostic biomarkers in AH. Here, we performed a comparative analysis of data derived from human hepatic and serum proteome to identify and characterize complement protein signatures in severe AH (sAH). The quantity of multiple complement proteins was perturbed in liver and serum proteome of patients with sAH. Multiple complement proteins differentiated patients with sAH from those with alcohol cirrhosis (AC) or alcohol use disorder (AUD) and healthy controls (HCs). Serum collectin 11 and C1q binding protein were strongly associated with sAH and exhibited good discriminatory performance among patients with sAH, AC, or AUD and HCs. Furthermore, complement component receptor 1-like protein was negatively associated with pro-inflammatory cytokines. Additionally, lower serum MBL associated serine protease 1 and coagulation factor II independently predicted 90-day mortality. In summary, meta-analysis of proteomic profiles from liver and circulation revealed complement protein signatures of sAH, highlighting a complex perturbation of complement and identifying potential diagnostic and prognostic biomarkers for patients with sAH.

Humans↗

PubMed2006

Altered expression of genes regulating skeletal muscle mass in the portacaval anastomosis rat.

We examined the temporal relationship between portacaval anastomosis (PCA), weight gain, changes in skeletal muscle mass and molecular markers of protein synthesis, protein breakdown, and satellite cell proliferation and differentiation. Male Sprague-Dawley rats with end to side PCA (n=24) were compared with sham-operated pair-fed rats (n=24). Whole body weight, lean body mass, and forelimb grip strength were determined at weekly intervals. The skeletal muscle expression of the ubiquitin proteasome system, myostatin, its receptor (the activin 2B receptor) and its signal, cyclin-dependent kinase inhibitor (CDKI) p21, insulin-like growth factor (IGF)-I and its receptor (IGF-I receptor-alpha), and markers of satellite cell proliferation and differentiation were quantified. PCA rats did not gain body weight and had lower lean body mass, forelimb grip strength, and gastrocnemius muscle weight. The skeletal muscle expression of the mRNA of ubiquitin proteasome components was higher in PCA rats in the first 2 wk followed by a lower expression in the subsequent 2 wk (P<0.01). The mRNA and protein of myostatin, activin 2B receptor, and CDKI p21 were higher, whereas IGF-I and its receptor as well as markers of satellite cell function (proliferating nuclear cell antigen, myoD, myf5, and myogenin) were lower at weeks 3 and 4 following PCA (P < 0.05). We conclude that PCA resulted in uninhibited proteolysis in the initial 2 wk. This was followed by an adaptive response in the later 2 wk consisting of an increased expression of myostatin that may have contributed to reduced muscle protein synthesis, impaired satellite cell function, and lower skeletal muscle mass.

Activin Receptors, Type II↗

PubMed2006

Inhibition of aromatase improves nutritional status following portacaval anastomosis in male rats.

BACKGROUND/AIMS: The portacaval anastamosis (PCA) rat is a model of the nutritional and endocrine consequences of liver cirrhosis. We hypothesized that failure to gain body weight in the PCA rat was the consequence of low testosterone levels and will be reversed by 4-hydroxyandrostenedione, a specific aromatase inhibitor. METHODS: Male Sprague-Dawley rats subjected to an end-to-side PCA or sham surgery were administered either 4-hydroxyandrostenedione or vehicle. Food intake, food efficiency, and body weight were measured, animals sacrificed 3 weeks after surgery, visceral organs harvested and plasma sex steroids measured. Hepatic RNA was extracted and dot blots done to quantify the expression of sex hormone dependent enzymes 16alpha hydroxylase and 15beta hydroxylase. RESULTS: Growth, food intake, food efficiency and plasma testosterone were lower and plasma estradiol higher in PCA than sham rats. Hepatic expression of testosterone driven 16alpha hydroxylase was lower and estradiol driven 15beta hydroxylase higher in PCA rats. These alterations were reversed by 4-hydroxyandrostenedione. CONCLUSIONS: These observations demonstrate that use of aromatase inhibitor reverses the nutritional and endocrine abnormalities in the PCA rat and suggest that this approach may be useful in cirrhotic patients.

Animals↗

PubMed2004

Skeletal muscle atrophy is associated with an increased expression of myostatin and impaired satellite cell function in the portacaval anastamosis rat.

Proliferation and differentiation of satellite cells are critical in the regeneration of atrophied muscle following immobilization and aging. We hypothesized that impaired satellite cell function is responsible for the atrophy of skeletal muscle also seen in cirrhosis. Myostatin and insulin-like growth factor 1 (IGF1) have been identified to be positive and negative regulators, respectively, of satellite cell function. Using a rat model of cirrhosis [portacaval anastamosis (PCA)] and sham-operated controls, we examined the expression of myostatin, its receptor activinR2b, and its downstream messenger cyclin-dependent kinase inhibitor p21 (CDKI p21) as well as IGF1 and its receptor in the gastrocnemius muscle. Expression of PCNA, a marker of proliferation, and myogenic regulatory factors (myoD, myf5, and myogenin), markers of differentiation of satellite cells, were also measured. Real- time PCR for mRNA and Western blot assay for protein quantification were performed. PCA rats had lower body weight and gastrocnemius weight compared with sham animals (P < 0.05). PCNA and myogenic regulatory factors were lower in PCA rats (P < 0.05). Myostatin, activinR2b, and CDKI p21 were higher in the PCA animals (P < 0.05). The expression of IGF1 and its receptor was lower in liver and skeletal muscle of PCA animals (P < 0.05). These data suggest that skeletal muscle atrophy seen in the portacaval shunted rats is a consequence of impaired satellite cell proliferation and differentiation mediated, in part, by higher myostatin and lower IGF1 expression.

Animals↗

PubMed2004

Protein restriction in hepatic encephalopathy: necessary evil or illogical dogma?

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Diet, Protein-Restricted↗

PubMed2004

MARS--does it stand the test of time?

Artificial liver support systems have been tested for decades in the management of liver failure. Generally, after some promising results published as case series, the device either disappears or fails to show significant benefit in controlled trials. Recently, the molecular absorbent recycling systems (MARS) or extracorporeal albumin dialysis (ECAD) technique appears to have broken this trend. Responding to the title one could summarize by saying this technique so far has stood the test of time. Data in support of its use in acute liver failure (ALF) is still scant and difficult to assess. However, in a well known but not very well defined entity of acute on chronic liver failure (AOCLF) the ECAD technique has been shown to improve survival compared to a similar randomized control group receiving standard supportive therapy. This well tolerated liver support system has real potential for widespread application if further well designed multicenter clinical trials continue to support its effectiveness. Its future lies probably in the management of the moribund hospitalized patient on the transplant list awaiting a donor liver.

Albumins↗

PubMed2003

Role of fresh frozen plasma infusion in correction of coagulopathy of chronic liver disease: a dual phase study.

OBJECTIVE: Fresh frozen plasma infusions are commonly used to correct the prolonged prothrombin time in patients with advanced chronic liver disease. The aim of this study was to establish how frequently this treatment is effective in correcting this coagulopathy. METHODS: A split retrospective-prospective study design was employed. In the retrospective series, 80 patients were identified with prolongation of the prothrombin time who received fresh frozen plasma infusions. In the prospective arm, 20 patients were included. All patients had confirmed chronic liver disease and showed no response to vitamin K injections. None of the patients had evidence of disseminated intravascular coagulation. The indications for infusion of fresh frozen plasma, number of units administered, complications, and percentage of patients who corrected their prothrombin time to less than 3 s longer than control time were recorded. RESULTS: The majority of patients (75%) received 2-4 units of fresh frozen plasma. The mean prothrombin time was numerically improved by the infusion of 2-6 units of fresh frozen plasma. However, using correction to less than 3 s longer than control time as an endpoint, only 12.5% of the retrospective and 10% of the prospective study groups respectively had correction of their coagulopathy. Only one complication of infusion of plasma was noted during the course of the study. CONCLUSIONS: Our results reiterate previous observations made more than 45 yr ago, that fresh frozen plasma infusions using the number of units commonly employed in clinical practice infrequently correct the coagulopathy of patients with chronic liver disease. Higher volumes (6 or more units) may be more effective but are rarely employed.

Adult↗

PubMed2003

Role of gut bacteria in the therapy of hepatic encephalopathy with lactulose and antibiotics.

The pathogenesis of hepatic encephalopathy is mediated to a large extent by ammonia, mainly derived from the gut. Both bacterial and nonbacterial mechanisms of ammoniagenesis have been shown, but ammoniagenesis mediated by colonic bacteria is probably of clinical significance. The therapy of hepatic encephalopathy is based on the putative etiological agents, including ammonia. Non-absorbable disaccharides and antibiotics have been shown to modify gut flora and decrease blood ammonia levels, but these are not necessarily related (indicating nonbacterial sources of ammonia, which may also be decreased by these compounds). A combination of these has been suggested but not consistently demonstrated to be beneficial in hepatic encephalopathy. Sodium benzoate is an alternate method of nonbacterial, non-hepatic metabolic binding pathway for ammonia disposal. Other mechanisms of gut bacterial modification may be achieved albeit transiently by the use of resistant bacteria like Enterococcus faecium.

Ammonia↗

PubMed2002

Preservation of portal pressure improves growth and metabolic profile in the male portacaval-shunted rat.

The portacaval anastomosis (PCA) rat model and human cirrhosis have many metabolic and nutritional abnormalities in common, such as growth retardation, hepatic and gonadal atrophy, and hyperammonemia. The severity of these abnormalities is variable and may be related to a number of factors, including portal pressure, portosystemic shunting, dietary intake, and how efficiently food is used. Therefore, this rat model was used to study these variables with the intent of gaining insights for improving the management of portal hypertension and malnutrition in human cirrhosis. A nonsuture end-to-side PCA (N = 100) or sham surgery (N = 71) was performed in 100 male rats. Four weeks after surgery, body and organ weights, food intake, serum ammonia, and serum amino acids were measured at death. In a subgroup of rats, (sham 7; PCA 34) portal venous pressure, degree of portosystemic shunting, and organ and body weights were obtained at death. Growth, liver weight, and testes weight were decreased, ammonia levels were higher, and the ratios of branched chain to aromatic amino acid (BCAA/AAA) were lower in the PCA group compared to the sham animals (P < 0.05). Since spleen weights correlated with portal pressure (P = 0.01), the PCA animals were then divided into those with preserved and those with low portal pressures based on spleen weight. The PCA group with preserved portal pressure had better growth, larger livers and testes, lower serum ammonia, and higher BCAA/AAA levels than the PCA group with low portal pressure; improvements associated with normal amounts of food intake and better food efficiency than the low pressure animals (P < 0.05 or better). Sham animals had no portosystemic shunting, while 100% shunting occurred in both PCA groups regardless of the portal pressure. In conclusion, preservation of portal pressure after portacaval anastomosis provides metabolic and nutritional benefits, which are independent of portosystemic shunting and associated with normal dietary intake and better preserved food efficiency.

Amino Acids, Aromatic↗

PubMed2001

Recent Advances in Schistosomiasis.

Schistosomiasis is a major parasitic disease, affecting nearly 200 million persons, worldwide. Major advances in our knowledge-in terms of pathogenesis, improved diagnosis, therapeutics (both drugs and strategies), and morbidity assessment-now make schistosomiasis a curable, often preventable disease. In contrast to most other illnesses, most schistosomiasis pathology appears to be reversible over time. For the future, several promising vaccine candidates are already in phase-I or phase-II testing. On the other hand, the range of this disease has been increasing, as water resources are developed in several newly industrialized countries and much of schistosomiasis in sub-Saharan Africa remains largely untreated.

Journal Article↗

PubMed2001

Hepatic Encephalopathy.

A characteristic feature of the neuropsychiatric changes in hepatic encephalopathy (HE) is the potential for a complete recovery in the majority of patients. In this review, we limit our discussion to HE in individuals with chronic liver disease. The optimal approach to the management of HE includes the following elements. Provide standard supportive therapy for patients with an altered mental status. This is the mainstay of therapy in the majority of clinical situations and includes administration of parenteral fluids and nutrition, care of vascular and bladder catheters, control of self-injurious activities, and instituting aspiration precautions. Rule out or control concomitant causes of encephalopathy. The diagnosis of HE has positive and negative criteria, and ruling out other causes of change in mental status is an essential component of the diagnosis. Identify and correct the precipitating factors of HE. In the majority of patients with HE, a clearly defined precipitating factor usually is identified, and the reversal or control of these factors is a key step in management. Institute gut-cleansing and ammonia-lowering measures. These measures are based on clinical and pathogenic characteristics of HE and are aimed at neutralizing the putative encephalogenic toxins (namely ammonia).

Journal Article↗