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Hairpin protein partitioning from the ER to lipid droplets involves major structural rearrangements.Nature Communications May 2024Lipid droplet (LD) function relies on proteins partitioning between the endoplasmic reticulum (ER) phospholipid bilayer and the LD monolayer membrane to control cellular...
Lipid droplet (LD) function relies on proteins partitioning between the endoplasmic reticulum (ER) phospholipid bilayer and the LD monolayer membrane to control cellular adaptation to metabolic changes. It has been proposed that these hairpin proteins integrate into both membranes in a similar monotopic topology, enabling their passive lateral diffusion during LD emergence at the ER. Here, we combine biochemical solvent-accessibility assays, electron paramagnetic resonance spectroscopy and intra-molecular crosslinking experiments with molecular dynamics simulations, and determine distinct intramembrane positionings of the ER/LD protein UBXD8 in ER bilayer and LD monolayer membranes. UBXD8 is deeply inserted into the ER bilayer with a V-shaped topology and adopts an open-shallow conformation in the LD monolayer. Major structural rearrangements are required to enable ER-to-LD partitioning. Free energy calculations suggest that such structural transition is unlikely spontaneous, indicating that ER-to-LD protein partitioning relies on more complex mechanisms than anticipated and providing regulatory means for this trans-organelle protein trafficking.
Topics: Endoplasmic Reticulum; Molecular Dynamics Simulation; Lipid Droplets; Electron Spin Resonance Spectroscopy; Humans; Lipid Bilayers; Protein Transport; Animals; Lipid Droplet Associated Proteins
PubMed: 38802378
DOI: 10.1038/s41467-024-48843-8 -
International Journal of Pharmaceutics Jun 2024In this study, Cannabidiol crystals (CBD) were used as a BCS class II model drug to generate a novel therapeutic deep eutectic solvent (THEDES) with easy preparation...
In this study, Cannabidiol crystals (CBD) were used as a BCS class II model drug to generate a novel therapeutic deep eutectic solvent (THEDES) with easy preparation using caprylic acid (CA). The hydrogen bonding interaction was confirmed by different techniques such as FT-IR and NMR, resulting in a hydrophobic system suitable for liquid formulations. The CBD-based THEDES, combined with a specific mixture of surfactants and co-surfactants, successfully formed a self-emulsifying drug delivery system (SEDDS) that generated uniform nano-sized droplets once dispersed in water. Hence, the THEDES showed compatibility with the self-emulsifying approach, offering an alternative method to load drugs at their therapeutic dosage. Physical stability concerns regarding the unconventional oily phase were addressed through stress tests using multiple and dynamic light scattering, demonstrating the robustness of the system. In addition, the formulated SEDDS proved effective in protecting CBD from the harsh acidic gastric environment for up to 2 h at pH 1.2. Furthermore, in vitro studies have confirmed the safety of the formulation and the ability of CBD to permeate Caco-2 cells when formulated. This investigation highlights the potential incorporation of THEDES in lipid-based formulations like SEDDS, expanding the avenues for innovative oral drug delivery approaches.
Topics: Caco-2 Cells; Humans; Emulsions; Solvents; Drug Delivery Systems; Cannabidiol; Caprylates; Surface-Active Agents; Hydrophobic and Hydrophilic Interactions; Drug Stability; Chemistry, Pharmaceutical; Emulsifying Agents
PubMed: 38797251
DOI: 10.1016/j.ijpharm.2024.124267 -
Viruses Apr 2024Rotavirus (RV) replicates within viroplasms, membraneless electron-dense globular cytosolic inclusions with liquid-liquid phase properties. In these structures occur the... (Review)
Review
Rotavirus (RV) replicates within viroplasms, membraneless electron-dense globular cytosolic inclusions with liquid-liquid phase properties. In these structures occur the virus transcription, replication, and packaging of the virus genome in newly assembled double-layered particles. The viroplasms are composed of virus proteins (NSP2, NSP5, NSP4, VP1, VP2, VP3, and VP6), single- and double-stranded virus RNAs, and host components such as microtubules, perilipin-1, and chaperonins. The formation, coalescence, maintenance, and perinuclear localization of viroplasms rely on their association with the cytoskeleton. A stabilized microtubule network involving microtubules and kinesin Eg5 and dynein molecular motors is associated with NSP5, NSP2, and VP2, facilitating dynamic processes such as viroplasm coalescence and perinuclear localization. Key post-translation modifications, particularly phosphorylation events of RV proteins NSP5 and NSP2, play pivotal roles in orchestrating these interactions. Actin filaments also contribute, triggering the formation of the viroplasms through the association of soluble cytosolic VP4 with actin and the molecular motor myosin. This review explores the evolving understanding of RV replication, emphasizing the host requirements essential for viroplasm formation and highlighting their dynamic interplay within the host cell.
Topics: Rotavirus; Virus Replication; Cytoskeleton; Humans; Animals; Microtubules; Viral Proteins; Host-Pathogen Interactions; Viral Nonstructural Proteins; Viral Replication Compartments; Rotavirus Infections; RNA, Viral
PubMed: 38793550
DOI: 10.3390/v16050668 -
International Journal of Molecular... May 2024Lipid droplet (LD) accumulation in hepatocytes is one of the major symptoms associated with fatty liver disease. Mitochondria play a key role in catabolizing fatty acids...
Lipid droplet (LD) accumulation in hepatocytes is one of the major symptoms associated with fatty liver disease. Mitochondria play a key role in catabolizing fatty acids for energy production through β-oxidation. The interplay between mitochondria and LD assumes a crucial role in lipid metabolism, while it is obscure how mitochondrial morphology affects systemic lipid metabolism in the liver. We previously reported that cilnidipine, an already existing anti-hypertensive drug, can prevent pathological mitochondrial fission by inhibiting protein-protein interaction between dynamin-related protein 1 (Drp1) and filamin, an actin-binding protein. Here, we found that cilnidipine and its new dihydropyridine (DHP) derivative, 1,4-DHP, which lacks Ca channel-blocking action of cilnidipine, prevent the palmitic acid-induced Drp1-filamin interaction, LD accumulation and cytotoxicity of human hepatic HepG2 cells. Cilnidipine and 1,4-DHP also suppressed the LD accumulation accompanied by reducing mitochondrial contact with LD in obese model and high-fat diet-fed mouse livers. These results propose that targeting the Drp1-filamin interaction become a new strategy for the prevention or treatment of fatty liver disease.
Topics: Animals; Dynamins; Humans; Lipid Droplets; Mice; Hep G2 Cells; Liver; Dihydropyridines; Mitochondria; Lipid Metabolism; Male; Mitochondrial Dynamics; Mice, Inbred C57BL; Diet, High-Fat; Hepatocytes
PubMed: 38791484
DOI: 10.3390/ijms25105446 -
Antioxidants (Basel, Switzerland) Apr 2024During our search for natural resources that can inhibit lipid droplet accumulation (LDA) and potentially prevent metabolic dysfunction-associated fatty liver disease...
During our search for natural resources that can inhibit lipid droplet accumulation (LDA) and potentially prevent metabolic dysfunction-associated fatty liver disease (MAFLD) and its progressive stages, such as metabolic dysfunction-associated steatohepatitis (MASH), eight bean extracts (BE1-BE8) were tested for their ability to inhibit lipid accumulation and oxidation in hepatocytes. Substantial inhibitory effects on LDA with bean extracts (BEs) BE2, BE4, BE5, and BE8 were demonstrated. An advanced lipidomic approach was used to quantify the accumulation and inhibition of intracellular triacylglycerol (TAG) and its oxidized species, TAG hydroperoxide (TGOOH), in hepatocytes under fatty acid-loading conditions. The results show that the antioxidants BE2 and BE8 are potential candidates for regulating TAG and TGOOH accumulation in fatty acid-induced lipid droplets (LDs). This study suggests that bean-based foods inhibit LDs formation by decreasing intracellular lipids and lipid hydroperoxides in the hepatocytes. The metabolic profiling of BEs revealed that BE2 and BE8 contained polyphenolic compounds. These may be potential resources for the development of functional foods and drug discovery targeting MAFLD/MASH.
PubMed: 38790618
DOI: 10.3390/antiox13050513 -
Gels (Basel, Switzerland) May 2024Edible hydrogel coatings or films in comparison to conventional food packaging materials are characterized as thin layers obtained from biopolymers that can be applied...
Edible hydrogel coatings or films in comparison to conventional food packaging materials are characterized as thin layers obtained from biopolymers that can be applied or enveloped onto the surface of food products. The use of lipid-containing hydrogel packaging materials, primarily as edible protective coatings for food applications, is recognized for their excellent barrier capacity against water vapor during storage. With the high brittleness of waxes and the oxidation of different fats or oils, highly stable agents are desirable. Jojoba oil obtained from the jojoba shrub is an ester of long-chain fatty acids and monovalent, long-chain alcohols, which contains natural oxidants α, β, and δ tocopherols; therefore, it is resistant to oxidation and shows high thermal stability. The production of hydrogel films and coatings involves solvent evaporation, which may occur in ambient or controlled drying conditions. The study aimed to determine the effect of drying conditions (temperature from 20 to 70 °C and relative humidity from 30 to 70%) and jojoba oil addition at the concentrations of 0, 0.5, 1.0, 1.5, and 2.0% on the selected physical properties of hydrogel edible films based on whey protein isolate. Homogenization resulted in stable, film-forming emulsions with bimodal lipid droplet distribution and a particle size close to 3 and 45 µm. When higher drying temperatures were used, the drying time was much shorter (minimum 2 h for temperature of 70 °C and relative humidity of 30%) and a more compact structure, lower water content (12.00-13.68%), and better mechanical resistance (3.48-3.93 MPa) of hydrogel whey protein films were observed. The optimal conditions for drying hydrogel whey protein films are a temperature of 50 °C and an air humidity of 30% over 3 h. Increasing the content of jojoba oil caused noticeable color changes (total color difference increased from 2.00 to 2.43 at 20 °C and from 2.58 to 3.04 at 70 °C), improved mechanical elasticity (the highest at 60 °C from 48.4 to 101.1%), and reduced water vapor permeability (the highest at 70 °C from 9.00·10 to 6.35·10 g/m·s·Pa) of the analyzed films. The observations of scanning electron micrographs showed the heterogeneity of the film surface and irregular distribution of lipid droplets in the film matrix.
PubMed: 38786257
DOI: 10.3390/gels10050340 -
Cells May 2024The possibility of detecting the developmental competence of individually cultured embryos through analysis of spent media is a major current trend in an ART setting....
The possibility of detecting the developmental competence of individually cultured embryos through analysis of spent media is a major current trend in an ART setting. However, individual embryo culture is detrimental compared with high-density group culture due to the reduced concentration of putative embryotropins. The main aim of this study was to identify an individual culture system that is not detrimental over high-density group culture in the bovine model. Blastocyst rates and competence were investigated in a conventional (GC) group, semi-confined group (MG), and individual culture (MS) in a commercial microwell device. Main findings showed that: (1) individual embryos can be continuously cultured for 7 days in ~70 nL microwells (MS) without detrimental effects compared with the GC and MG; (2) MS and MG blastocysts had a reduced number of TUNEL-positive cells compared to GC blastocysts; (3) though blastocyst mean cell numbers, mitochondrial activity, and lipid content were not different among the three culture conditions, MS blastocysts had a higher frequency of small-sized lipid droplets and a reduced mean droplet diameter compared with GC and MG blastocysts. Overall, findings open the way to optimize the development and competence of single embryos in an ART setting.
Topics: Animals; Cattle; Blastocyst; Zygote; Embryo Culture Techniques; Embryonic Development; Female; Mitochondria
PubMed: 38786090
DOI: 10.3390/cells13100868 -
Cells May 2024In recent decades, there has been a dramatic rise in the rates of children being born after in utero exposure to drugs of abuse, particularly opioids. Opioids have been...
In recent decades, there has been a dramatic rise in the rates of children being born after in utero exposure to drugs of abuse, particularly opioids. Opioids have been shown to have detrimental effects on neurons and glia in the central nervous system (CNS), but the impact of prenatal opioid exposure (POE) on still-developing synaptic circuitry is largely unknown. Astrocytes exert a powerful influence on synaptic development, secreting factors to either promote or inhibit synapse formation and neuronal maturation in the developing CNS. Here, we investigated the effects of the partial µ-opioid receptor agonist buprenorphine on astrocyte synaptogenic signaling and morphological development in cortical cell culture. Acute buprenorphine treatment had no effect on the excitatory synapse number in astrocyte-free neuron cultures. In conditions where neurons shared culture media with astrocytes, buprenorphine attenuated the synaptogenic capabilities of astrocyte-secreted factors. Neurons cultured from drug-naïve mice showed no change in synapses when treated with factors secreted by astrocytes from POE mice. However, this same treatment was synaptogenic when applied to neurons from POE mice, indicating a complex neuroadaptive response in the event of impaired astrocyte signaling. In addition to promoting morphological and connectivity changes in neurons, POE exerted a strong influence on astrocyte development, disrupting their structural maturation and promoting the accumulation of lipid droplets (LDs), suggestive of a maladaptive stress response in the developing CNS.
Topics: Astrocytes; Animals; Synapses; Female; Pregnancy; Mice; Analgesics, Opioid; Prenatal Exposure Delayed Effects; Neurons; Signal Transduction; Buprenorphine; Cells, Cultured; Mice, Inbred C57BL
PubMed: 38786059
DOI: 10.3390/cells13100837 -
Biomolecules May 2024Epidemiological and clinical evidence have extensively documented the role of obesity in the development of endometrial cancer. However, the effect of fatty acids on...
Epidemiological and clinical evidence have extensively documented the role of obesity in the development of endometrial cancer. However, the effect of fatty acids on cell growth in endometrial cancer has not been widely studied. Here, we reported that palmitic acid significantly inhibited cell proliferation of endometrial cancer cells and primary cultures of endometrial cancer and reduced tumor growth in a transgenic mouse model of endometrial cancer, in parallel with increased cellular stress and apoptosis and decreased cellular adhesion and invasion. Inhibition of cellular stress by N-acetyl-L-cysteine effectively reversed the effects of palmitic acid on cell proliferation, apoptosis, and invasive capacity in endometrial cancer cells. Palmitic acid increased the intracellular formation of lipid droplets in a time- and dose-dependent manner. Depletion of lipid droplets by blocking DGAT1 and DGAT2 effectively increased the ability of palmitic acid to inhibit cell proliferation and induce cleaved caspase 3 activity. Collectively, this study provides new insight into the effect of palmitic acid on cell proliferation and invasion and the formation of lipid droplets that may have potential clinical relevance in the treatment of obesity-driven endometrial cancer.
Topics: Female; Palmitic Acid; Endometrial Neoplasms; Humans; Lipid Droplets; Animals; Cell Proliferation; Mice; Apoptosis; Cell Line, Tumor; Diacylglycerol O-Acyltransferase; Mice, Transgenic
PubMed: 38786008
DOI: 10.3390/biom14050601 -
Biomolecules May 2024Here, we describe GS-9, a novel water-soluble fatty acid-based formulation comprising L-lysine and arachidonic acid, that we have shown to induce ferroptosis. GS-9 forms...
Here, we describe GS-9, a novel water-soluble fatty acid-based formulation comprising L-lysine and arachidonic acid, that we have shown to induce ferroptosis. GS-9 forms vesicle-like structures in solution and mediates lipid peroxidation, as evidenced by increased C11-BODIPY fluorescence and an accumulation of toxic malondialdehyde, a downstream product of lipid peroxidation. Ferroptosis inhibitors counteracted GS-9-induced cell death, whereas caspase 3 and 7 or MLKL knock-out cell lines are resistant to GS-9-induced cell death, eliminating other cell death processes such as apoptosis and necroptosis as the mechanism of action of GS-9. We also demonstrate that through their role of sequestering fatty acids, lipid droplets play a protective role against GS-9-induced ferroptosis, as inhibition of lipid droplet biogenesis enhanced GS-9 cytotoxicity. In addition, Fatty Acid Transport Protein 2 was implicated in GS-9 uptake. Overall, this study identifies and characterises the mechanism of GS-9 as a ferroptosis inducer. This formulation of arachidonic acid offers a novel tool for investigating and manipulating ferroptosis in various cellular and anti-cancer contexts.
Topics: Ferroptosis; Arachidonic Acid; Humans; Lipid Peroxidation; Cell Line, Tumor; Water; Solubility; Neoplasms; Lipid Droplets
PubMed: 38785962
DOI: 10.3390/biom14050555