Oxidized phospholipid-protein adducts: The future targets of interest.

Phospholipids are key biomolecules with important roles as components of membranes, lipoproteins and as signalling molecules. However, phospholipids are quite prone to oxidation. Upon oxidation they generate several types of oxidation products including long chain oxidation products, as hydroperoxyl and hydroxy derivatives, and highly reactive oxidation products, like small aldehydes and truncated oxidized phospholipids. The formation of protein adducts with small electrophilic aldehydes (like malondialdehyde) is now well studied, however, the aggregation of proteins with truncated oxidized phospholipids lacks research. This paper provides a short overview of the formation of protein adducts with truncated oxidized phospholipids as well as a gathering of the research on this topic. The literature found reports the synthesis, detection and fragmentation of this type of adducts, mainly focusing on truncated oxidized phospholipid' products from phosphatidylcholine class and few peptides and proteins, as human serum albumin and Apo B100, leaving unattended the screening in vivo and in disease correlation, thus lacking possible association with their biological role. These adducts are a consequence of oxidative modifications to important biomolecules and their involvement in the organism is still unclear, revealing the urgent need for more investigation in this area.

Whole Blood and Plasma-Based Lipid Profiling Reveals Distinctive Metabolic Changes in Systemic Lupus Erythematosus and Systemic Sclerosis.

Autoimmune diseases (AID), such as systemic lupus erythematosus (SLE) and systemic sclerosis (SS), are complex conditions involving immune system dysregulation. Diagnosis is challenging, requiring biomarkers for improved detection and prediction of relapses. Lipids have emerged as potential biomarkers due to their role in inflammation and immune response. This study uses an untargeted C18 RP-LC-MS lipidomics approach to comprehensively assess changes in lipid profiles in patients with SLE and SS. By analyzing whole blood and plasma, the study aims to simplify the lipidomic analysis, explore cellular-level lipids, and compare lipid signatures of SLE and SS with healthy controls. Our findings showed variations in the lipid profile of SLE and SS. Sphingomyelin and ceramide molecular species showed significant increases in plasma samples from SS patients, suggesting an atherosclerotic profile and potentially serving as lipid biomarkers. Phosphatidylserine species in whole blood from SLE patients exhibited elevated levels supporting previously reported dysregulated processes of cell death and defective clearance of dying cells in this AID. Moreover, decreased phospholipids bearing PUFA were observed, potentially attributed to the degradation of these species through lipid peroxidation processes. Further studies are needed to better understand the role of lipids in the pathological mechanisms underlying SLE and SS.

Lipid profile variability in children at different ages measured in dried blood spots.

Dried blood spot (DBS) is a minimally invasive sampling technique that has several advantages over conventional venipuncture/arterial blood sampling. More recently, DBS has also been applied for lipidomics analysis, but this is an area that requires further research. The few works found in the literature on lipidomics of DBS samples performed the analysis in adult samples, leaving pediatric ages unmapped. The objective of this study was to assess the variability of the lipid profile (identified by high-resolution C18 RP-LC-MS/MS) of DBS at pediatric age (0-10 days, 2-18 months, and 3-13 years) and to identify age-related variations. The results revealed that the lipidomic signature of the three age groups is significantly different, especially for a few species of neutral lipids and phosphatidylcholines. The main contributors to the differentiation of the groups correspond to 3 carnitine (Car), 2 cholesteryl ester (CE), 2 diacylglycerol (DG), 2 triacylglycerol (TG), 3 phosphatidylcholine (PC), 1 ether-linked PC, 1 phosphatidylethanolamine (PE), 1 ether-linked PE and 1 phosphatidylinositol (PI) species, all with statistically significant differences. Additionally, lipid species containing linoleic acid (C18:2) were shown to have significantly lower levels in the 0-10 days group with a gradual increase in the 2-18 month, reaching the highest concentrations in the 3-13 year group. The results of this study highlighted the adaptations of the lipid profile at different pediatric ages. These results may help improve understanding of the evolution of lipid metabolism throughout childhood and should be investigated further.

Dried blood spots in clinical lipidomics: optimization and recent findings.

Dried blood spots (DBS) are being considered as an alternative sampling method of blood collection that can be used in combination with lipidomic and other omic analysis. DBS are successfully used in the clinical context to collect samples for newborn screening for the measurement of specific fatty acid derivatives, such as acylcarnitines, and lipids from whole blood for diagnostic purposes. However, DBS are scarcely used for lipidomic analysis and investigations. Lipidomic studies using DBS are starting to emerge as a powerful method for sampling and storage in clinical lipidomic analysis, but the major research work is being done in the pre- and analytical steps and procedures, and few in clinical applications. This review presents a description of the impact factors and variables that can affect DBS lipidomic analysis, such as the type of DBS card, haematocrit, homogeneity of the blood drop, matrix/chromatographic effects, and the chemical and physical properties of the analyte. Additionally, a brief overview of lipidomic studies using DBS to unveil their application in clinical scenarios is also presented, considering the studies of method development and validation and, to a less extent, for clinical diagnosis using clinical lipidomics. DBS combined with lipidomic approaches proved to be as effective as whole blood samples, achieving high levels of sensitivity and specificity during MS and MS/MS analysis, which could be a useful tool for biomarker identification. Lipidomic profiling using MS/MS platforms enables significant insights into physiological changes, which could be useful in precision medicine.

Understanding the nitrolipidome: From chemistry to mass spectrometry and biological significance of modified complex lipids.

Complex lipids, phospholipids (PLs) and triacylglycerides (TAGs), are prone to modifications induced by reactive nitrated species and reactive oxygen species, generating a range of nitrated, nitrosated or nitroxidized derivatives, as nitro PLs and nitro TAGs. These modified lipids (epilipids) have been reported in vitro and in vivo using lipidomics approaches. However, their detection in living systems remains a challenge hampered by its complexity, high structural diversity, and low abundance. The advances in high-resolution mass spectrometry combined with the higher sensitivity of the instruments like Orbitrap-based mass spectrometers opened new opportunities for the detection of these modified complex lipids. This review summarizes the challenges and findings behind the identification of nitrated, nitrosated and nitroxidized PLs and TAGs fragmentation fingerprints based on collision-induced dissociation (CID) and higher energy CID (HCD) MS/MS approaches. Following what has already been reported for nitrated fatty acids, these complex lipids are found to act as endogenous mediators with potential electrophilic properties and can express bioactivities such as anti-inflammatory and antioxidant actions. This information can be used to design untargeted and targeted lipidomics strategies for these modified complex lipids in biological samples as well as in pathological, food and industrial settings, further unveiling their biological and signalling roles.

Looking in Depth at Oxidized Cholesteryl Esters by LC-MS/MS: Reporting Specific Fragmentation Fingerprints and Isomer Discrimination.

Cholesteryl esters (CE) are prone to oxidation under increased oxidative stress conditions, but little is known about oxidized CE species (oxCE). To date, only a few oxCE have been identified, however, mainly based on the detection of molecular ions by mass spectrometry (MS) or target approaches for specific oxCE. The study of oxCE occurring from radical oxidation is still scarcely addressed. In this work, we made a comprehensive assessment of oxCE derivatives and their specific fragmentation patterns to identify detailed structural features and isomer differentiation using high-resolution C18 HPLC-MS- and MS/MS-based lipidomic approaches. The LC-MS/MS analysis allowed us to pinpoint oxCE structural isomers of long-chain and short-chain species, eluting at different retention times (tR). Data analysis revealed that oxCE can be modified either in the fatty acyl moiety or in the cholesterol ring. The location of the hydroxy/hydroperoxy group originates characteristic fragment ions, namely the unmodified cholestenyl cation (m/z 369) for the isomer with oxidation in the fatty acyl chain or ions at m/z 367 and m/z 385 (369 + 16) when oxygenation occurs in the cholesterol ring. Additionally, we identified CE 18:2 and 20:4 aldehydic and carboxylic short-chain products that showed a clear fragmentation pattern that confirmed the modification in the fatty acyl chain. Specific fragmentation fingerprinting allowed discrimination of the isobaric short-chain species, namely carboxylic short-chain products, from hydroxy aldehyde short-chain products, with a hydroxycholesterol moiety. This new information is important to identify different oxCE in biological samples and will contribute to unraveling their role in biological conditions and diseases such as cardiovascular disease.

Insights in the Role of Lipids, Oxidative Stress and Inflammation in Rheumatoid Arthritis Unveiled by New Trends in Lipidomic Investigations.

Rheumatoid arthritis (RA) is a highly debilitating chronic inflammatory autoimmune disease most prevalent in women. The true etiology of this disease is complex, multifactorial, and is yet to be completely elucidated. However, oxidative stress and lipid peroxidation are associated with the development and pathogenesis of RA. In this case, oxidative damage biomarkers have been found to be significantly higher in RA patients, associated with the oxidation of biomolecules and the stimulation of inflammatory responses. Lipid peroxidation is one of the major consequences of oxidative stress, with the formation of deleterious lipid hydroperoxides and electrophilic reactive lipid species. Additionally, changes in the lipoprotein profile seem to be common in RA, contributing to cardiovascular diseases and a chronic inflammatory environment. Nevertheless, changes in the lipid profile at a molecular level in RA are still poorly understood. Therefore, the goal of this review was to gather all the information regarding lipid alterations in RA analyzed by mass spectrometry. Studies on the variation of lipid profile in RA using lipidomics showed that fatty acid and phospholipid metabolisms, especially in phosphatidylcholine and phosphatidylethanolamine, are affected in this disease. These promising results could lead to the discovery of new diagnostic lipid biomarkers for early diagnosis of RA and targets for personalized medicine.

Serum phospholipidomics reveals altered lipid profile and promising biomarkers in multiple sclerosis.

Multiple sclerosis is a neurodegenerative disease causing disability in young adults. Alterations in metabolism and lipid profile have been associated with this disease. Several studies have reported changes in the metabolism of arachidonic acid and the profile of fatty acids, ceramides, phospholipids and lipid peroxidation products. Nevertheless, the understanding of the modulation of circulating lipids at the molecular level in multiple sclerosis remains unclear. In the present study, we sought to assess the existence of a distinctive lipid signature of multiple sclerosis using an untargeted lipidomics approach. It also aimed to assess the differences in lipid profile between disease status (relapse and remission). For this, we used hydrophilic interaction liquid chromatography coupled with mass spectrometry for phospholipidomic profiling of serum samples from patients with multiple sclerosis. Our results demonstrated that multiple sclerosis has a phospholipidomic signature different from that of healthy controls, especially the PE, PC, LPE, ether-linked PE and ether-linked PC species. Plasmalogen PC and PE species, which are natural endogenous antioxidants, as well as PC and PE polyunsaturated fatty acid esterified species showed significantly lower levels in patients with multiple sclerosis and patients in both remission and relapse of multiple sclerosis. Our results show for the first time that the serum phospholipidome of multiple sclerosis is significantly different from that of healthy controls and that few phospholipids, with the lowest p-value, such as PC(34:3), PC(36:6), PE(40:10) and PC(38:1) may be suitable as biomarkers for clinical applications in multiple sclerosis.

An overview of lipidomic analysis in different human matrices of multiple sclerosis.

Multiple sclerosis is a chronic inflammatory and neurodegenerative disease of the central nervous system, and it is one of the most common neurological cause of disability in young adults. It is known that several factors contribute to increase the risk of development and pathogenesis of multiple sclerosis, nonetheless, but the true etiology of this pathology remains unknown. Similar to other inflammatory diseases, oxidative stress and lipid peroxidation are also associated to multiple sclerosis. Alterations in the lipid profile seem to be a hallmark of this pathology which can contribute to the dysregulation of lipid homeostasis and lipid metabolism in multiple sclerosis. Lipidomic studies analysed in this review clearly demonstrate the role of lipids in inflammatory processes, in immunity, and in the onset and development of multiple sclerosis. Several investigations reported alterations of some molecular lipid species, in particular, with decrease of fatty acids (FA) 18:2 and 20:4 and total polyunsaturated FA, with compensatory increases of saturated FA with shorter carbon chains. Oxidized phospholipids were reported in few studies as well. Also, it was shown that clinical lipidomics has potential as a tool to aid both in multiple sclerosis diagnosis and therapeutics by allowing a detailed lipidome profiling of the patients suffering with this disease.

Lipidomics in autoimmune diseases with main focus on systemic lupus erythematosus.

Autoimmune diseases (AID) are a heterogeneous group of disorders that have in common a chronic inflammation and dysregulation of the immune system. Systemic lupus erythematosus (SLE) is one of the most frequent systemic autoimmune diseases characterized by autoimmune phenomena in multiple organs. The tests used for evolution and prognosis assessment are either non-specific or non-sensitive, impairing an adequate therapeutics. To face this drawback, lipidomics is being used to provide more knowledge and insights regarding autoimmune disorders. Through lipidomic approaches using MS, it is possible to identify and quantify the level of lipid molecular species in the biological system and this could be useful to identify biomarkers and to better understand the pathophysiology of autoimmune diseases. There are some evidence that lipids and oxidized lipids can play a key role in AID pathogenesis. Although this field has been scarcely explored, there are some studies that reported variations on the lipid profile at a molecular level using lipidomic approaches based on MS in SLE. The results gathered herein showed changes mainly in the level of phospholipids, with decrease of some plasmenyl lipids, fatty acids, with reduction of PUFA, and sphingolipids, with changes in fatty acyl chain composition. These changes may be the result of lipids` modifications due to oxidation and increase of ROS. Some alterations can be associated with changes in membrane of lymphocytes and with the deregulation of the immune system. Thus, exploring the knowledge from modern lipidomic approaches in the study of the role of lipids and oxidized lipids, in oxidative stress and in inflammatory diseases, could contribute for the identification of new lipid biomarkers. Lipid biomarkers are promising tools to prognosis and treatment monitoring, tailored for the best therapeutic response and highest safety to ensure better patient care and to be used for personalized medicine.