Transcriptomic Profiling of L-Proline Induced Ferroptosis Reveals PPARγ-Mediated Regulatory Networks in HFpEF Progression
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE308324 | GEO | 2025/09/17
REPOSITORIES: GEO
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Project description:Ferroptosis is a form of regulated cell death driven by the iron-dependent accumulation of oxidized polyunsaturated-fatty-acid-containing phospholipids (PUFA-PLs). Three key molecular features of ferroptosis are peroxidation of PUFA-PLs, accumulation of redox-active ferrous iron, and defective lipid peroxide repair (Dixon and Stockwell, 2019). However, there is currently no reliable way to selectively stain ferroptotic cells in tissue sections to characterize the extent of ferroptosis in animal models of disease and in patient tissue samples. We sought to address this gap by immunizing mice with membranes from lymphoma cells treated with the ferroptosis inducer piperazine erastin (PE), and screening ~4,750 of the resulting monoclonal antibodies generated for their ability to selectively detect cells undergoing ferroptosis. We found that one antibody, termed 3F3 Ferroptotic Membrane Antibody (3F3-FMA), was effective as a selective ferroptosis-staining reagent using immunofluorescence (IF). The antigen of 3F3-FMA was identified by immunoprecipitation and mass spectrometry as the human transferrin receptor 1 protein (TfR1), which imports iron from the extracellular environment into cells. We validated this finding with several additional anti-TfR1 antibodies, and compared them to other potential ferroptosis-detecting reagents via immunofluorescence staining, using fluorescence microscopy and flow cytometry. We found that anti-TfR1 and anti-malondialdehyde (MDA) adduct antibodies were effective at reliably staining ferroptotic tumor cells in numerous cell culture and tissue contexts. In summary, these findings suggest that anti-TfR1 antibodies can be used to selectively label cells undergoing ferroptosis.
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Project description:Ferroptotic cell death is dependent on unrestricted lipid peroxidation rather than activation of apoptotic effector caspases. A large number of ferroptosis activators have been reported recently. We used gene sequencing to analyze the effects of four ferroptosis activators (RSL3, N6F11, Fin56 and Erastin) on global gene expression in human PDAC1 cell line.
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Project description:In this study the authors used systems biology to define progressive changes in metabolism and transcription in a large animal model of heart failure with preserved ejection fraction (HFpEF). Transcriptomic analysis of cardiac tissue, 1 month post-banding, revealed loss of electron transport chain components, and this was supported by changes in metabolism and mitochondrial function, altogether signifying alterations in oxidative metabolism. Established HFpEF, 4 months post-banding, resulted in changes in intermediary metabolism with normalized mitochondrial function. Mitochondrial dysfunction and energetic deficiencies were noted in skeletal muscle at early and late phases of disease, suggesting cardiac-derived signaling contributes to peripheral tissue maladaptation in HFpEF. Collectively, these results provide insights into the cellular biology underlying HFpEF progression.
2021-09-01 | GSE181184 | GEO
Project description:Although the prevalence of heart failure with preserved ejection fraction (HFpEF) is increasing, evidence-based therapies for HFpEF are rare, likely due to an incomplete understanding of this disease. This study sought to identify the cardiac-specific features of protein and phosphoprotein in a murine model of HFpEF using mass spectrometry. HFpEF mice developed moderate hypertension, left ventricle (LV) hypertrophy, lung congestion and diastolic dysfunction. Proteomics analysis of the LV tissue showed that 897 proteins were differentially expressed between HFpEF and Sham mice. We observed abundant changes in sarcomeric proteins, mitochondrial-related proteins, and NAD-dependent protein deacetylase sirtuin-3 (SIRT3). Upregulated pathways by GSEA analysis were related to immune modulation and muscle contraction, while downregulated pathways were predominantly related mitochondrial metabolism. Western blot analysis validated SIRT3 downregulated cardiac expression in HFpEF. Phosphoproteomics analysis showed that 72 phosphopeptides were differentially regulated between HFpEF and sham LV. Aberrant phosphorylation patterns mostly occurred in sarcomere proteins and nuclear-localized proteins associated with contractile dysfunction and cardiac hypertrophy. Seven aberrant phospho-sites were observed at the z disk binding region of titin. While total titin cardiac expression remained unaltered, its stiffer N2B isoform was significantly increased in HFpEF. Thus, these results show profound changes in proteins related to mitochondrial metabolism and function and the cardiac contractile apparatus in HFpEF. We propose that SIRT3 plays a key role and may be a target for drug development in HFpEF.
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Project description:To investigate the effect of butyrate on cardiac transcripts in western-diet induced pre-HFpEF
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2025-02-28 | GSE260465 | GEO
Project description:Metabolic reprogramming is critical in the onset of pressure overload-induced cardiac remodeling. Our study reveals that proline dehydrogenase (PRODH), the key enzyme in proline metabolism, reprograms cardiomyocyte metabolism to protect against cardiac remodeling. We induced cardiac remodeling using transverse aortic constriction (TAC) in both cardiac-specific PRODH knockout and overexpression mice. Our results indicate that PRODH expression is suppressed post-TAC. Cardiac-specific PRODH knockout mice exhibited worsened cardiac dysfunction, while mice with PRODH overexpression demonstrated a protective effect. Additionally, we simulated cardiomyocyte hypertrophy in vitro using neonatal rat ventricular myocytes treated with phenylephrine. Through RNA sequencing, metabolomics, and metabolic flux analysis, we elucidated that PRODH overexpression in cardiomyocytes redirects proline catabolism to replenish tricarboxylic acid (TCA) cycle intermediates, enhance energy production, and restore glutathione redox balance. In summary, our findings suggest PRODH as a modulator of cardiac bioenergetics and redox homeostasis duing cardiac remodeling induced by pressure overload. This highlights the potential of PRODH as a therapeutic target for cardiac remodeling.
2025-02-28 | GSE260464 | GEO
Project description:One of the most critical axes for cell fate determination is how cells respond to excessive reactive oxygen species (ROS)-oxidative stress. Extensive lipid peroxidation commits cells to death via a distinct cell death paradigm termed ferroptosis. However, the molecular mechanism regulating cellular fates to distinct ROS remains incompletely understood. Through siRNA against human receptor-interacting protein kinases (RIPK) family members, we discovered that RIPK4 is crucial for oxidative stress and ferroptotic death. Upon ROS induction, RIPK4 is rapidly activated and the kinase activity of RIPK4 is indispensable to induce cell death. Specific ablation of RIPK4 in kidney proximal tubules protects mice from acute kidney injury induced by cisplatin and renal ischemia/reperfusion. RNA sequencing revealed the dramatically decreased expression of acyl-CoA synthetase medium-chain (ACSM) family members induced by cisplatin treatment which is compromised in RIPK4 deficient mice. Among these ACSM family members, suppression of ACSM1 strongly augments oxidative stress and ferroptotic cell death with induced expression of ACSL4, an important component for ferroptosis execution. Our lipidome analysis revealed that over-expression of ACSM1 leads to the accumulation of monounsaturated fatty acids (MUFAs), attenuation of polyunsaturated fatty acids (PUFAs) production, and thereby cellular resistance to ferroptosis. Hence, knock-down ACSM1 re-sensitizes RIPK4 KO cells to oxidative stress and ferroptotic death. In conclusion, RIPK4 is a key player involved in oxidative stress and ferroptotic death, which is potentially important for a broad spectrum of human pathologies. The link between RIPK4-ASCM1 axis to PUFAs and ferroptosis reveals a novel mechanism to oxidative stress induced necrosis and ferroptosis.
2024-09-04 | GSE276038 | GEO
Project description:Increased protein acetylation is frequently observed in the failing heart, including in hearts with heart failure with preserved ejection fraction (HFpEF). However, the role of protein acetylation in cardiac metabolic impairments during the pathogenesis of HFpEF remains insufficiently investigated. In this study, a two-hit strategy, involving a high-fat diet and Nω-nitro-L-arginine methyl ester (L-NAME), was employed to induce the HFpEF model in mice. Significant cardiac diastolic dysfunction, pathological remodeling, and enhanced protein hyperacetylation were observed in the hearts of the two-hit HFpEF mice. Acetylome profiling revealed that the hyperacetylated proteins were predominantly localized to mitochondria and enriched in metabolic pathways, particularly in fatty acid oxidation (FAO). Furthermore, the HFpEF heart exhibited reduced FAO capacity and abnormal lipid accumulation. Activation of mitochondrial protein deacetylation restored FAO capacity and alleviated cardiac dysfunction in the HFpEF heart, whereas inhibition of mitochondrial deacetylase directly induced lipid metabolism disorders in cardiomyocytes. Notably, we identified Dlat, a pyruvate metabolism enzyme, as the key transacetylase responsible for mitochondrial protein hyperacetylation in the HFpEF heart. Overexpression of Dlat enhanced FAO-related protein acetylation and exacerbated cardiac lipid metabolism disturbances, thereby inducing pathological changes resembling the HFpEF phenotype. In contrast, Dlat knockdown effectively mitigated FAO inhibition and functional impairment in the two-hit HFpEF mice. Through discovery-driven approaches, we demonstrated that Dlat directly binds to the alpha subunit of mitochondrial trifunctional protein (HADHA) and triggers its acetylation at the K728 site, thereby inactivating HADHA enzymatic activity. Reactivating HADHA, either by cardiac-specific HADHA overexpression or oral administration of a biogenic polyamine, restored cardiac FAO capacity and prevented the development of HFpEF. Our study provides a mechanistic basis linking protein hyperacetylation, FAO inhibition, and the development of HFpEF. Activation of mitochondrial deacetylase or enhancement of cardiac FAO capacity may offer novel strategies for restoring cardiac metabolic homeostasis and function in HFpEF.
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Project description:<p>Ferroptosis is a form of regulated cell death with roles in degenerative diseases and cancer. Excessive iron-catalyzed peroxidation of membrane phospholipids, especially those containing the polyunsaturated fatty acid arachidonic acid (AA), is central in driving ferroptosis. Here, we reveal that an understudied Golgi-resident scaffold protein, MMD, promotes susceptibility to ferroptosis in ovarian and renal carcinoma cells in an ACSL4- and MBOAT7-dependent manner. Mechanistically, MMD physically interacts with both ACSL4 and MBOAT7, two enzymes that catalyze sequential steps to incorporate AA in phosphatidylinositol (PI) lipids. Thus, MMD increases the flux of AA into PI, resulting in heightened cellular levels of AA-PI and other AA-containing phospholipid species. This molecular mechanism points to a pro-ferroptotic role for MBOAT7 and AA-PI, with potential therapeutic implications, and reveals that MMD is an important regulator of cellular lipid metabolism.</p>
2023-09-06 | MTBLS8280 | MetaboLights