Protein glycosylation governs key physiological and pathological processes in human cells. Aberrant glycosylation is thus closely associated with disease progression. Mass spectrometry (MS)-based glycoproteomics has emerged as an indispensable tool for investigating glycosylation changes in biological samples with high sensitivity. Following rapid improvements in methodologies for reliable intact glycopeptide identification, site-specific quantification of glycopeptide macro- and micro-heterogeneity at the proteome scale has become an urgent need for exploring glycosylation regulations. Here, we summarize recent advances in N- and O-linked glycoproteomic quantification strategies and discuss their limitations. We further describe a strategy to propagate MS data for multilayered glycopeptide quantification, enabling a more comprehensive examination of global and site-specific glycosylation changes. Altogether, we show how quantitative glycoproteomics methods explore glycosylation regulation in human diseases and promote the discovery of biomarkers and therapeutic targets. Glycosylation occurs ubiquitously on vital biomacromolecules, including proteins, lipids and nucleic acids . In partThis article summarizes current MS-based technologies for proteome-wide quantification of N- and O-linked protein glycosylation . These t13C, 15N, 18O and 2H, to label glycopeptides. Such labeling creates mass shifts to glycopeptide precursors or fragmentation products so that glycopeptides from different samples can be differently labeled and pooled before measurement but remain distinguishable by mass spectrometry. Stable isotopes can be incorporated chemically, metabolically or enzymatically into amino acids or glycans on glycopeptides , isobaric tags for relative and absolute quantitation (iTRAQ), isotope-encoded dimethylated leucine or isobaric N, N-dimethyl leucine (DiLeu) and isobaric tag (IBT), permit global glycopeptide quantification with high sample multiplexity  ,74,75,76Rapidly improving software packages further speeds up the applications of isobaric labeling to quantitative glycoproteomics. For example, Byonic, one of the pioneering software programs developed for large-scale identification of both N- and O-linked glycopeptides , can worGPQuest, developed by the Zhang group, also allows for confident identification and isobaric-labeling-based quantification of intact N-glycopeptides ,22,23,79Stadlmann et al. also developed a comparative glycoproteomics platform combining a new N-glycopeptide identification algorithm, SugarQb, with the quantitative workflows embedded in Proteome Discoverer . They usMultiplexed isobaric labeling reduces the overall LC-MS measurement time and variations introduced between measurements ,75. In aAlong the same line, Zhu et al. combined electron-transfer dissociation (ETD) with a standard multi-notch workflow, resulting in improved identification and quantification of TMT-labeled intact glycopeptides . Using aIn summary, rapidly developing MS technologies and software tools enhance the quality of intact glycopeptide analysis and enable more straightforward data analysis for non-experts. Although commercial isobaric labeling reagents may not be affordable to all due to the expensive costs, recent developments have significantly promoted their applications in quantitative glycoproteomics.In contrast to isobaric labeling, isotopic chemical labeling allows the quantification of intact N-/O-glycopeptides at the MS1 level. Its multiplexing capacity is often limited to up to three  due to the increased MS1 spectrum complexity and overlapping glycopeptide isotopic peaks. Labeling is commonly performed after proteolysis and followed by equal mixing. After enrichment, differently labeled glycopeptides co-elute chromatographically and appeared distinguishable in the MS1 spectra. The relative peak intensities/areas of the light and heavy-labeled glycopeptides are compared.12CD2O) or 6n Da (13CD2O) mass difference, where n is the number of amino groups in peptides. The Tian group developed a dedicated software GPSeeker and GPSeekerQuan for the identification and quantification of dimethyl labeled intact N-glycopeptides [3XHO, X = 13C or 12C) and NaBH3CN, for the relative quantitation of intact N-glycopeptides with enhanced accuracy and dynamic range [Dimethyl labeling is based on the reaction of peptide primary amines (N-terminus or ϵ-amino group of Lys residue of a peptide) with formaldehyde, introducing 4n  into endogenous proteins metabolically during cell culture [The stable isotope labeling by amino acids in cell culture (SILAC) method incorporates  culture . DiffereBenjamin et al. performed quantitative N-glycomics and SILAC-based quantitative N-glycoproteomics to study protein glycosylation changes in adipocytes upon TNF-alpha-induced insulin resistance . They obAlthough SILAC reduced quantitation errors introduced during sample preparation, it is not applicable to many biological materials, especially to patient tissues, and has a limited number  of conditions to be compared in one experiment. Micro-heterogeneous glycoforms on one peptide core have closely related masses and do not separate well on the standard C18 chromatography, which often makes MS1 spectra of intact glycopeptide analysis more complicated. SILAC inherently further increases the MS1 complexity, resulting in interfered SILAC pair determination and chromatographic ion extraction. The introduced SILAC pair can also cause the over-sequencing of the same glycopeptides and undersampling of all available glycopeptides in DDA analysis.18O to the former N-glycosylated asparagine residues when reacting in an 18O-labeled buffer. The +2 Da mass shift enables glycosite identification and glycosylation occupancy determination [16O- and 18O-labeled peptides, Liu et al. developed tandem 18O stable isotope labeling (TOSIL), in which three 18O atoms are incorporated into de-glycosylated peptides for determining N-glycosylation site occupancy [18O atoms into the carboxyl termini of all peptides during tryptic digestion and a third 18O atom into the N-glycosylation site while cleaving glycans by PNGase F. Thus, compared to normal 16O de-glycopeptides, a mass shift of 6 Da for singly glycosylated peptides or 8 Da for doubly glycosylated peptides appears in MS1 scans. The intensity ratio of 18O/16O isotopic peaks represents the abundance difference between the glycosylated peptide  and its non-glycosylated counterpart, allowing for determining changes in N-glycosylation site occupancy [PNGase F efficiently releases N-glycans and introduces one mination ,90,91. Tccupancy . The TOSccupancy ,42.18O labeling methods for the relative quantification of intact glycopeptides in serum from patients with hepatitis-B-virus-related liver diseases [18O atoms into the carboxyl termini of intact glycopeptides during tryptic digestion, resulting in a +4 Da mass difference in mass spectrometry. A combination of pGlyco [Recently, Zhang et al. further applied the diseases . They inf pGlyco  and pQuaf pGlyco  was used18O to 16O during sample preparation. In addition, the overlapped isotopic peaks of 18O- and 16O-labeled glycopeptides (with only a 4 Da mass difference) also complicate the data processing steps for quantification.Enzymatic labeling is a relatively simple and cost-effective approach for quantitative glycoproteomics. However, downstream data analysis should consider the back exchange of In addition to the peptide backbones, stable isotopes can also be incorporated into glycans attached to glycopeptides. The isotopic methylamine (MeSIL) method labels the carboxyl groups on both the sialic acid residues and the peptide moiety  at the same time . Labeled15N-enriched or 13C-enriched media can replace all the N and C atoms in glycopeptides with 15N and 13C, respectively. This method was successfully applied to yeast N-glycoproteomics. Liu et al. applied the 15N and 13C labeling to yeast glycopeptides to evaluate the true false discovery rate (FDR) of pGlyco [15N- or 13C-labeled yeast proteins were pooled and then analyzed in one LC-MS/MS run. The detected mass difference between an unlabeled glycopeptide and its 15N or 13C-labeled counterpart should be equal to the number of nitrogen or carbon atoms calculated based on the assigned glycopeptide composition. Accordingly, they filtered out the false identifications and determined the true FDR. To the best of our knowledge, pQuant is the only software supporting quantifying glycopeptides bearing 15 N- and 13C-labeled glycans.Alternatively, metabolic labeling using f pGlyco . Equal aBiocompatible azido-bearing monosaccharides are commonly used for metabolic glycan labeling ,94. The Label-free quantification (LFQ) methods aim at quantifying glycopeptides without the use of stable-isotope labels. The samples to be compared were prepared separately, including protein extraction, digestion, glycopeptide enrichment and parallel LC-MS/MS measurements. The extracted ion currents (XIC) or spectral counts of identified glycopeptides across runs were compared. Depending on the MS acquisition methods, the LFQ can be classified into data-dependent acquisition (DDA) and data-independent acquisition (DIA) based quantification . In the DDA-based label-free quantification uses either the MS1 intensities or the spectral counts of identified glycopeptides for simple and accurate glycopeptide quantification. Benefitting from the rapidly developing software packages, numerous data processing tools support intact glycopeptide LFQ analysis , including LFQuant , iBAQ 4, ByologiGlycosylation heterogeneity makes the LFQ of intact glycopeptides more troublesome than the standard proteomic analysis. Glycoforms sharing the same peptide sequence can have close masses and elute closely in a narrow retention time range, resulting in overlapping glycopeptide signals and thus interfering with glycopeptide peak extraction. Nevertheless, by combining the intensities of the top three isotope peaks at the three highest MS spectral points, Integrated GlycoProteome Analyzer (I-GPA) enables fast and sensitive glycopeptide LFQ . AlthougDDA analysis is inherently biased against low-abundance glycopeptides, which either do not produce sufficient detectable fragment ions for reliable identification or are not selected for fragmentation at all. Limited by the scan speed of available mass spectrometers, it is common that many existing glycopeptides are not identified in all DDA runs, resulting in missing values for LFQ. Although numerous existing proteomics data processing methods allow assigning and matching MS1 features across all raw files to minimize LFQ missing values ,102,103,An alternative method to MS-intensity-based DDA quantification is spectral counting, which utilizes counts of identified glycopeptide-to-spectrum matches (GPSMs) to represent the abundance of glycopeptides. Because of the random-selection and intense-peak-first triggering strategy of a DDA analysis, more abundant glycopeptides in theory lead to higher numbers of GPSMs than low-abundance peptides. This method was recently applied to quantify the site-specific glycosylation of SARS-CoV-2 spike protein and human ACE2 receptor . AdditioWith faster mass spectrometers and advanced data processing tools, data-independent acquisition (DIA) methods offering improved sensitivity and reproducibility compared traditional DDA are rising in quantitative proteomics . This apA DIA analysis of intact N-glycopeptides is much more complex than analyzing de-glycopeptides, requiring certain adjustments and the optimization on LC-MS settings  and on data analysis pipelines . As one of the pioneers, Zacchi et al. applied DIA-based N-glycoproteomics to study the changes in glycan macro- and micro-heterogeneity in mature proteins caused by mutations in the N-glycosylation pathway in Saccharomyces cerevisiae . They crSimilarly, Sanda and Goldman also reported a SWATH workflow for detecting IgG glycoforms from human plasma. However, instead of b- and y-ions, they used Y-ions generated under lower collision energy for quantification. These manually curated Y-ions with a high yield of up to 60% precursor intensity were proven to be highly specific to each glycoform . In a foMore recently, Ye et al. developed a DIA-based strategy for quantitative O-GalNAc–type glycoproteomic analysis in complex biological samples . They fiIn general, the confident identification of glycopeptides requires several levels of information: peptide sequence identification, glycan moiety identification and, ideally, glycosite localization. Numerous studies have elucidated strategies to estimate the false discovery rate (FDR) of glycopeptides in a DDA analysis . HoweverIn summary, DDA-based glycopeptide LFQ, either using MS1 feature intensity or spectral count, has the advantage of simple workflows and lower cost but requires sophisticated data processing methods, retention time alignment, and post-acquisition normalization to account for the MS response variations in replicate measurements. This approach also suffers from severe missing values in large-scale glycoproteomics due to drastic differences in glycoform abundances, and low identification rates of less abundant glycopeptides in DDA. DIA-based LFQ methods have shown the potential to quantitate intact glycopeptides with higher sensitivity and fewer missing values. However, current glycopeptide DIA analysis still relies on a pre-established, DDA-generated spectral library, limiting its application. The so-called directDIA approach and the use of a predicted spectral library are not yet available in the glycoproteome field . NeverthIn targeted acquisition methods, such as selected or multiple reaction monitoring (SRM or MRM) or parallel reaction monitoring (PRM), LC-MS assays are deployed to detect glycopeptides of interest with high sensitivity, reproducibility and quantitative accuracy. SRM analysis was initially developed on triple quadrupole instruments, where the first quadrupole (Q1) scans for a pre-defined precursor, which undergoes CID fragmentation in the second quadrupole (Q2). Subsequently, pre-defined ions among the resulting fragments are scanned in the third quadrupole (Q3) A.To achieve robust and sensitive glycopeptide detection in an SRM/MRM analysis, transition  selection is crucial. Depending on the fragmentation modes, various types of glycopeptide fragments can be selected as target transitions, including B ions (oxonium ions), Y ions  and b/y ions (peptide fragment ions without glycans) B. MonitoThe major drawback of targeted acquisition is that the number of precursor ions to be monitored is limited by the scan speed of the mass spectrometer. N-linked glycopeptides typically display heterogeneous glycoforms that elute within a narrow retention time window, resulting in overlapping LC peaks. Given that robust quantification requires at least 6–8 data points across a chromatographic peak, glycosylation heterogeneity often overwhelms the MS instrument duty cycle.As mentioned above, various quantitative glycoproteomics strategies determine glycosylation changes at different levels C. For inWe recently proposed the SugarQuant workflow for multiplexed quantitative site-specific N-glycoproteomics . In the Glycosylation plays critical roles in protein functions and cellular processes, and its aberration is closely associated with human diseases, including cancers , neurodeAlpha-fetoprotein (AFP) and core fucosylated AFP-L3 are closely correlated with hepatocellular carcinoma (HCC) and have been successfully used as clinical biomarkers for HCC diagnosis. The AFP level shows a huge variation among HCC patient populations. By relatively quantifying the intact N-glycopeptides between low and high levels of AFP tumor groups, several sialylated but not core fucosylated tri-antennary glycans were uniquely increased in HCC tumors with low AFP level, while many core fucosylated bi-antennary or hybrid glycans as well as bisecting glycans were uniquely increased in tumors with high AFP . Other in = 10/grade) and BPH tissues (n = 5) revealed several protein-, cell- and tumor-grade-specific N- and O-glycosylation [Prostate-specific antigen (PSA) is a US Food and Drug Administration (FDA)-approved serum biomarker for prostate cancer (PCa) screening, and its glycosylation profiles in PCa patients provide the potential for improving the specificity of the PSA test ,132. A Hsylation . Quantitsylation . They fusylation .Glycoproteomics-based signatures can be used for tumor subtyping and clinical outcome prediction in ovarian cancer and provide potential for precision medicine . Zhang’sAs nicely summarized by Khan and Cabral, the aberrant glycosylation of markers of cancer stem cells (CSCs) in key cellular signaling pathways has been directly correlated with the self-renewal properties and drug-resistant mechanisms . The TiaProtein glycosylation alteration is also associated with other cancers, including breast cancer, pancreatic cancer and gastric cancer. Fang et al. identified a group of 11 glycopeptides from serum mannose receptor as a potential marker for differentiating and stratifying breast cancer subtypes . AnotherIn addition to cancers, dysregulated N-glycosylation occurs in neurological and cardiac disorders. For instance, recent N-glycoproteomic studies of brains or cerebrospinal fluid obtained from patients or mice with Alzheimer’s disease revealed highly heterogeneous and dysregulated protein N-glycosylation alterations ,124,138.Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused the COVID-19 pandemic. The heavily glycosylated spike (S) protein located on the SARS-CoV-2 surface binds to human angiotensin-converting enzyme 2 (ACE2) and mediates host-cell entry . GlycansHowever, it is crucial to note that recombinant S proteins showed different site-specific glycosylation than those originating from wild-type SARS-CoV-2 virions . FurtherIn addition to the S protein, its binding partner, the ACE2 receptor, is heavily glycosylated, too. All seven glycosylation sites in hACE2 were found to be completely occupied, mainly by complex N-glycans ,149. IntGlycoproteomics methods are maturing along with the ever-improving algorithms for confident and fast glycopeptide identification at the proteome scale. These methods allow researchers to uncover the glycosylation regulations and the underlying mechanisms in disease development. However, the inherent complexity of glycosylation limits the direct application of the well-developed quantitative methods for large-scale intact glycopeptide quantification. There are numerous technical considerations when planning a quantitative glycoproteomics study, such as labeling efficiency, the mass difference among the isotope-coded labels, variations introduced by glycopeptide enrichment and the available data processing tools. Distinct methods can significantly differ in quantitative performance, sensitivity, throughput and cost. This article summarizes the currently applicable approaches for quantitative glycoproteomics and discusses their advantages and disadvantages individually .In general, isobaric chemical labeling provides unprecedented multiplexing capability (up to 21 channels), decreasing both instrument time and run-to-run variation. Ion mobility and MS3-based reporter quantification can efficiently reduce the ratio distortion caused by co-isolation interference and improve quantitative performance. In contrast, metabolic labeling allows sample pooling at the very first step of sample preparation, avoiding technical variations introduced during sample preparation. However, limited by available isotope-coded reagents, metabolic labeling often introduces a mass shift on the labeled glycopeptides, which complicates the MS1 scans. Overlapping isotopic peaks of the labeled glycopeptides can further lead to biased quantification.Label-free quantification enables a straightforward and cost-effective sample preparation workflow but is hungrier for instrument time and more vulnerable to run-to-run variation. DIA analysis is still in its infancy for large-scale glycopeptide quantification. It improves the missing value and reproducibility but is also more challenging for proper data analysis. On the other hand, target methods provide higher sensitivity and are the methods of choice for validating potential biomarkers and clinical applications.We also summarize recent applications of quantitative glycoproteomics to explore the macro- and micro-heterogeneity of glycopeptides in various diseases and reveal glycosylation regulation in disease progression. Such analyses often involve the integration of multiple methods  to construct a global view of glycosylation regulation. To ease the technical challenges, we proposed a streamlined approach for obtaining multi-layered glycosylation quantification in a single experiment. Glycopeptide analysis has also contributed to studying the glycan shield on proteins of coronavirus strains that cause the ongoing pandemic. We anticipate that rapidly developing MS technologies and software tools will bring about more accurate, precise, sensitive and user-friendly quantification methods for intact glycopeptides, accelerating biomarker discovery and therapeutic target identification.


In December 2020, the Food and Drug Administration (FDA) issued Emergency Use Authorizations (EUAs) for the Pfizer-BioNTech COVID-19 (BNT162b2) vaccine and the Moderna COVID-19 (mRNA-1273) vaccine,Since June 2020, ACIP has convened 15 public meetings to review data on COVID-19 epidemiology and use of COVID-19 vaccines. The ACIP COVID-19 Vaccines Work Group, comprising experts in infectious diseases, vaccinology, vaccine safety, public health, and ethics, has held weekly meetings since April 2020 to review COVID-19 surveillance data, evidence for vaccine efficacy and safety, and implementation considerations for COVID-19 vaccination programs. After reports of myocarditis, the work group met twice to review clinical trial and postauthorization safety data for myocarditis after receipt of mRNA COVID-19 vaccines. The work group also reviewed a benefit-risk assessment of myocarditis events after receipt of mRNA COVID-19 vaccines, considering recent epidemiology of COVID-19 and sequelae of COVID-19, including myocarditis and multisystem inflammatory syndrome in children (MIS-C).Myocarditis typically occurs more commonly in males than in females, and incidence is highest among infants, adolescents, and young adults . SupportAs of June 11, 2021, approximately 296 million doses of mRNA COVID-19 vaccines had been administered in the United States, with 52 million administered to persons aged 12–29 years; of these, 30 million were first and 22 million were second doses. Within the Vaccine Adverse Event Reporting System (VAERS) . The medUsing myocarditis cases reported to VAERS with onset within 7 days after dose 2 of an mRNA vaccine, crude reporting rates  per million second dose recipients were calculated using national COVID-19 vaccine administration data as of June 11, 2021. Myocarditis reporting rates were 40.6 cases per million second doses of mRNA COVID-19 vaccines administered to males aged 12−29 years and 2.4 per million second doses administered to males aged ≥30 years; reporting rates among females in these age groups were 4.2 and 1.0 per million second doses, respectively.To assess the benefit-risk balance of mRNA vaccines in adolescents and young adults, ACIP reviewed an individual-level assessment that compared the benefits  to the risks (number of cases of myocarditis) of vaccination, using methods similar to those described previously.The benefits  outweighed the risks (expected myocarditis cases after vaccination) in all populations for which vaccination has been recommended. However, the balance of benefits and risks varied by age and sex because cases of myocarditis were primarily identified among males aged <30 years, and the risks of poor outcomes related to COVID-19 increase with age. Per million second doses of mRNA COVID-19 vaccine administered to males aged 12–29 years, 11,000 COVID-19 cases, 560 hospitalizations, 138 ICU admissions, and six deaths due to COVID-19 could be prevented, compared with 39–47 expected myocarditis cases after COVID-19 vaccination . Among mACIP also reviewed population-level considerations regarding vaccination. No alternatives to mRNA COVID-19 vaccines for adolescents will be available for the foreseeable future, and vaccination of adolescents offers protection against COVID-19 that can be important for returning to educational, social, and extracurricular activities. Higher levels of vaccination coverage can reduce community transmission, which can protect against development and circulation of emerging variants. Regarding health equity considerations, racial and ethnic minority groups have higher rates of COVID-19 and severe diseaseThe ACIP discussion concluded that 1) the benefits of vaccinating all recommended age groups with mRNA COVID-19 vaccine clearly outweigh the risks of vaccination, including the risk of myocarditis after vaccination; 2) continuing to monitor outcomes of myocarditis cases after COVID-19 vaccination is important; and 3) providers and the public should be informed about these myocarditis cases and the use of COVID-19 vaccines. Based on ACIP’s conclusion regarding the benefit-risk assessment on June 23, 2021, COVID-19 vaccination continues to be recommended for all persons aged ≥12 years under the FDA’s EUA. ACIP emphasized the importance of informing vaccination providers and the public about the benefits and the risks, including the risk for myocarditis after COVID-19 vaccination, particularly for males aged 12–29 years.https://www.cdc.gov/vaccines/covid-19/clinical-considerations/myocarditis.html), as well as considerations for a second vaccine dose in persons who develop myocarditis after a first dose . FDA has added information to the Pfizer-BioNTechCDC has provided guidance regarding evaluation and management of myocarditis after mRNA COVID-19 vaccine  that uses text messaging and online surveys to provide near real-time health check-ins after receipt of a COVID-19 vaccine. In cases of v-safe reports that include possible medically attended health events, CDC’s v-safe call center follows up with the vaccine recipient to collect additional information for completion of a VAERS report. Information on v-safe is available at https://www.cdc.gov/vsafe.FDA requires that vaccine providers report to VAERS vaccination administration errors, serious adverse events,An elevated risk for myocarditis among mRNA COVID-19 vaccinees has been observed, particularly in males aged 12–29 years.On June 23, 2021, the Advisory Committee on Immunization Practices concluded that the benefits of COVID-19 vaccination to individual persons and at the population level clearly outweighed the risks of myocarditis after vaccination.Continued use of mRNA COVID-19 vaccines in all recommended age groups will prevent morbidity and mortality from COVID-19 that far exceed the number of cases of myocarditis expected. Information regarding the risk for myocarditis with mRNA COVID-19 vaccines should be disseminated to providers to share with vaccine recipients.


Hyperalphalipoproteinemia (HALP) is a lipid disorder characterized by elevated plasma high-density lipoprotein cholesterol (HDL-C) levels above the 90th percentile of the distribution of HDL-C values in the general population. Secondary non-genetic factors such as drugs, pregnancy, alcohol intake, and liver diseases might induce HDL increases. Primary forms of HALP are caused by mutations in the genes coding for cholesteryl ester transfer protein (CETP), hepatic lipase (HL), apolipoprotein C-III (apo C-III), scavenger receptor class B type I (SR-BI) and endothelial lipase (EL). However, in the last decades, genome-wide association studies (GWAS) have also suggested a polygenic inheritance of hyperalphalipoproteinemia. Epidemiological studies have suggested that HDL-C is inversely correlated with cardiovascular (CV) risk, but recent Mendelian randomization data have shown a lack of atheroprotective causal effects of HDL-C. This review will focus on primary forms of HALP, the role of polygenic inheritance on HDL-C, associated risk for cardiovascular diseases and possible treatment options. Hyperalphalipoproteinemia (HALP) is a condition characterized by elevated plasma high-density lipoprotein cholesterol (HDL-C) levels > 90th percentile of the distribution of HDL-C values in the general population , associaHDLs are characterized by a heterogenous sub-population of lipoprotein particles, which undergo remodeling and transformation processes mediated by several plasma enzymes and transcription factors . The maiAlthough in this review we will mainly focus on the primary familial causes of HALP, it is worth mentioning that several conditions are known to be associated with elevated HDL-C levels. Patients with secondary HALP tend to be asymptomatic, aside from some rare reported cases of juvenile or premature corneal opacities  or multiThe CETP gene maps on the long arm of chromosome 16 16q12-16q21); it comprises 16 exons  and enco-16q21; iAPO-CIII. Apolipoprotein CIII (Apo-CIII) is a small apolipoprotein synthesized mainly in the liver and regulates plasma TG homeostasis by inhibiting lipoprotein lipase (LPL) activity [APOC3 gene (c.13-2A > G and c.55+1G > A) have been identified and associated with plasma HDL-C levels above the 95th percentile and an atheroprotective lipid profile [HL deficiency is another cause of monogenic HALP. Hepatic lipase (HL) is a heparan-sulfate proteoglycan (HSPG)-bound lipolytic enzyme synthesized and secreted by hepatocytes. It is encoded by the LIPC gene mapping on chromosome 15 , and it activity . Apo-CIIactivity ,85. Two  profile . A misse profile .The scavenger receptor class B type I (SR-BI), encoded by the SCARB1 gene, is primarily expressed in the steroidogenic tissues and in the liver, where it acts as an important receptor for HDLs and controls selective uptake of the cholesterol esters by HDL . SR-BI iEndothelial lipase (EL) is mostly involved in HDL phospholipid hydrolysis  through Several polymorphisms of the CETP gene with a reduction in CETP activity have been associated with HDL-C levels in the general population . TaqIB iFollowing the seminal data provided by Gofman J.W. et al.  and the The concept that targeting HDL-C may be advantageous in terms of CV risk reduction has been taken into consideration for decades as an important treatment strategy. Niacin is a potent HDL-C-raising drug, seemingly an attractive approach to reduce cardiac events in patients with or at risk of atherosclerotic cardiovascular disease . HoweverCETP inhibitors have been proposed over the years to raise HDL-C plasma levels, although clinical trials have failed to prove their hypothesized positive effects ,156. AmoClinical trials are investigating the role of new targets to evaluate cardioprotection by increasing specific HDL-C subclasses or improving HDL functions . ReconstIn summary, primary HALP is a heterogenous genetic lipid disorder and some pathogenic mechanisms are still not completely understood. The supposed protective role of HDL on CV risk have been extensively evaluated over the years, and several studies have shown that different mechanisms based on HDL-C increases by enhancing both the cholesterol efflux and the cholesterol esterification are not definitely anti-atherogenic. Moreover, the different metabolic scenarios and specific biochemical pathways governed by the numerous HDL-related genes need further studies in order to clarify in which way a similar HDL-C increase may induce atherogenic or athero-protective effects. New functional assays would be necessary to measure the HDL quality in a validated, reproducible and cost-effective manner to better understand the mechanisms related to cardiovascular protection.


The viewpoint proposes a carbon-neutral biochemical cycle in the cytosol of plant leaves that is up-regulated by reactive oxygen species. Cycling provides NADPH and dissipates energy to counteract oxidative stress. Leaf cytosol contains non-phosphorylating and phosphorylating glyceraldehyde-3-phosphate dehydrogenase . From the viewpoint of carbon metabolism, np-GAPDH is redundant. However, mutants lacking np-GAPDH show significant metabolic adjustments and decreased growth, suggesting that np-GAPDH has central functions in plant metabolism. Here, I propose a cytosolic oxidation–reduction cycle. In its forward direction, np-GAPDH supplies NADPH. In the reverse direction, phosphoglycerate kinase and p-GAPDH consume ATP and NADH. Thus, the cytosolic oxidation–reduction cycle may constitute a central hub in energy metabolism.+ carrier proteins in reconstituted systems show low affinities for this reductant  3PGA is reimported into chloroplasts by the triose phosphate translocator and reduced to dihydroxyacetone phosphate by chloroplastic phosphoglycerate kinase (PGK) and phosphorylating (p)-GAPDH. A similar cycle involving p-GAPDH and PGK in the forward direction was suggested to provide cytosolic NADH and ATP  cycle . In its  systems . Here, tThe cytosol of plant leaves contains two distinct GAPDHs. While np-GAPDH catalyses the irreversible conversion of GAP to 3PGA, p-GAPDH together with PGK catalyses the same reaction in a reversible manner . Thus, fAt the sequence level, np-GAPDH and p-GAPDH are entirely unrelated . Additionally, np-GAPDH from various sources has an ~10-fold higher affinity for GAP than p-GAPDH  due to drought. Low Cc promotes an energy excess in chloroplasts due to decreased consumption of ATP and NADPH by the Calvin–Benson cycle but continued electron input by light-harvesting complexes . Much of the cytosolic ATP is believed to derive from the mitochondrial oxidation of photorespiratory glycine . Excess NADPH in chloroplasts is shuttled out to the cytosol by the malate valve as NADH. Flux through the valve is regulated strictly by the activity of chloroplastic malate dehydrogenase, which increases with [NADPH]/[NADP+] ratios  catalysed by PGK and p-GAPDH require cytosolic ATP and NADH . This reReactive oxygen species inhibit p-GAPDH (see above). If all p-GAPDH were in its oxidized inactive state, COR cycling would halt. Hence, cycling requires retained p-GAPDH activity. COR cycling involves only three cytosolic enzymes and is carbon neutral. It requires no net carbon input, causes no net carbon loss, and does not produce carbon products that need to be consumed by other processes. Hence, COR cycling is independent of other parts of carbon metabolism as well as transmembrane transport of triose or pentose phosphates. This provides flexibility to its functions.In vivo, NADPH concentration is believed to exert primary control over np-GAPDH activity (see above). Thus, COR cycling may help to maintain NADPH concentrations at high levels to steadily support all NADPH-consuming processes in the cytosol.Optimal plant functioning requires a well-balanced energy supply versus consumption. Under most conditions, NADPH and ATP supply exceed metabolic demands, and several processes dissipating excess energy have been proposed . Export


As neurological complications associated with COVID-19 keep unfolding, the number of cases with COVID-19-associated de novo movement disorders is rising. Although no clear pathomechanistic explanation is provided yet, the growing number of these cases is somewhat alarming. This review gathers information from 64 reports of de novo movement disorders developing after/during infection with SARS-CoV-2. Three new cases with myoclonus occurring shortly after a COVID-19 infection are also presented. Treatment resulted in partial to complete recovery in all three cases. Although the overall percentage of COVID-19 patients who develop movement disorders is marginal, explanations on a probable causal link have been suggested by numerous reports; most commonly involving immune-mediated and postinfectious and less frequently hypoxic-associated and ischemic-related pathways. The current body of evidence points myoclonus and ataxia out as the most frequent movement disorders occurring in COVID-19 patients. Some cases of tremor, chorea, and hypokinetic-rigid syndrome have also been observed in association with COVID-19. In particular, parkinsonism may be of dual concern in the setting of COVID-19; some have linked viral infections with Parkinson's disease (PD) based on results from cerebrospinal fluid analyses, and PD is speculated to impact the outcome of COVID-19 in patients negatively. In conclusion, the present paper reviewed the demographic, clinical, and treatment-associated information on de novo movement disorders in COVID-19 patients in detail; it also underlined the higher incidence of myoclonus and ataxia associated with COVID-19 than other movement disorders. Coronavirus disease 2019 (COVID-19), a lower respiratory infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is associated with a wide range of neurological manifestations. Most frequent neurological symptoms are nonspecific and include headache, dizziness, disturbance of smell and taste, and myalgia . MovemenHypothetically, there are multiple entry routes for the virus into the CNS  . HoweverWe conducted a comprehensive systematic literature search in PubMed and Scopus databases using the following terms: “coronavirus disease 2019,” “COVID,” “COVID-19,” “SARS-CoV-2,” “Neurologic,” “Movement disorders,” “Tremor,” “Myoclonus,” “Parkinsonism,” “ataxia,” “dystonia,” and “chorea”. The Mendeley application was used to detect, scan, and remove duplicates. The remaining studies were scanned for relevant titles/abstracts. All relevant articles that reported patients who developed a movement disorder during/after a SARS-CoV-2 infection were included. Reference lists of the obtained articles were searched for additional relevant results. Any movement disorders in COVID-19 patients were reviewed; demographic information and data on clinical characteristics, treatments applied, and the outcome of patients were extracted. We included case reports, case series, and observational studies published in peer-reviewed journals and preprints available in English. Articles without full texts and studies without laboratory confirmation (RT-PCR) of COVID-19 diagnoses were excluded.Our extensive search in the literature yielded a final number of 43 publications reporting movement disorders in 64 patients with SARS-CoV-2 infection . DemograA previously healthy 42-year-old man presented with difficulty speaking, cognitive deficits, ataxia, and progressive disabling jerks in upper and lower limbs, leading to falls  . All sixInfection-associated myoclonus can often be encountered among patients with severe viral, bacterial, or parasitic infections and is characterized by abrupt, brief, and sometimes repetitive contractions of the trunk, limbs, or face muscles , 25. MyoTremor is another type of COVID-19-associated abnormal movement observed in hospitalized COVID-19 patients . We idenDespite having been rarely attributed to infections before this pandemic, numerous cases of cerebellar ataxia in COVID-19 patients (during or after their infection course) have raised concerns regarding a potential causal link probably through immune-mediated pathomechanisms , 54.The most recognized chorea associated with infections is Sydenham's chorea, characterized by autoimmunity against the basal ganglia after a streptococcal infection . We idenThe acute hypokinetic-rigid syndrome has been reported in two cases (cases no. 5  and 13 ); both dα-synuclein retrograde spreading and aggregation from the olfactory bulb to the midbrain and deeper CNS structures—leading to cellular death of the affected tissue—could offer some protection against SARS-CoV-2 breaching into the CNS via the olfactory route [Parkinson's disease (PD) is a neurodegenerative disease caused by the destruction of dopaminergic neurons in substantia nigra pars compacta (SNpc). Evidence of viral infections associated with PD exists; thus, vigilant monitoring and investigation of probable links between SARS-CoV-2 infection and parkinsonism may be necessary , 56. On ry route . Howeverry route , 9. Regary route . Furtherry route .A community-based case-control study on 12 PD patients who contracted COVID-19 documented a substantial risk of motor and nonmotor symptoms worsening associated with mild to moderate degrees of COVID-19, regardless of the patients' age and disease duration .Lo Monaco and colleagues reported a patient with an 8-year history of PD who developed severe generalized dystonia shortly after contracting COVID-19. Improvement was achieved after an increase in her daily dose of levodopa and clozapine .Inversely, PD and its associated disabilities may negatively impact the outcome of COVID-19 illness, along with other known risk factors such as age and hypertension. Indeed, Fasano et al. reported a higher COVID-19 mortality rate in community-dwelling PD patients  than the general population , and SalSome limitations apply to the present review article, such as the keywords selection; some studies that have not used the keywords incorporated in our search strategy, e.g., those that only mentioned “neurological manifestations” without further specification, might have been missed. The literature search has not thoroughly adhered to the standards of a systematic review, albeit being as exhaustive and comprehensive as possible. Furthermore, cross-reference checking for some reviewed articles may have been incomplete. Review papers bear the limitation of their reviewed articles; as discussed earlier, myoclonus may stem from metabolic comorbidities in some cases. Thus, the attribution of myoclonus to COVID-19 by some case studies is a potential limitation of such reports applying to the present review as well. Another limitation was that electrophysiological data were not obtained from two of our cases .This review highlighted the higher incidence of myoclonus and ataxia after COVID-19 infection. Other movement disorders were not common after COVID-19, implying that SARS-CoV-2 mainly affects the brain through immune-mediated pathways rather than a direct invasion of the CNS. Nonetheless, the COVID-19 pandemic can affect the outcome of patients with established movement disorders.


EGFR gene. Nevertheless, the 18.9-month median progression-free survival emphasizes the fact that resistance to osimertinib therapy is inevitable. Acquired resistance mechanisms to osimertinib in EGFR-driven NSCLC include MET amplification, EGFR C797S mutation, neuroendocrine differentiation, small-cell lung carcinoma histologic transformation, PD-L1 and KRAS amplifications and ESR1-AKAP12 and MKRN1-BRAF translocations, as well as BRAF V600 mutation. This last one represents 3% of the acquired resistance mechanisms to osimertinib. In this review, we discuss the rationale for EGFR/BRAF/MEK co-inhibition in the light of a clinical case of EGFR-mutant NSCLC developing a BRAF V600 mutation as an acquired resistance mechanism to osimertinib and responding to the association of osimertinib plus dabrafenib and trametinib. Additionally, we discuss the acquired resistance mechanisms to osimertinib plus dabrafenib and trametinib combination in that context.Osimertinib has become a standard of care in the first-line treatment of advanced-stage non-small-cell lung cancer (NSCLC) harboring exon 19 and 21 activating mutations in the  EGFR) and B-Raf proto-oncogene (BRAF) genes or translocations in anaplastic lymphoma kinase (ALK), Ret proto-oncogene (RET), or C-Ros-1 proto-oncogene (ROS1) genes [EGFR mutation is the leading genetic alteration in NSCLC, accounting for 12% of unselected lung adenocarcinomas (LUADs) and 44% of LUADs in never-smokers in the Caucasian population. Exon 19 deletion and exon 21 L858R sensitizing EGFR mutations constitute 86% of EGFR mutations in NSCLC and confer high responsiveness (sensitivity) to EGFR tyrosine kinase inhibitors (TKIs) [EGFR activating mutations significantly improved the outcome of EGFR-mutant NSCLC. Recently, a third-generation TKI, osimertinib, improved progression-free survival (PFS), overall survival (OS) and intracerebral efficacy in comparison to first-generation EGFR TKIs gefitinib or erlotinib in advanced-stage EGFR-mutant NSCLC [EGFR-mutant NSCLC.Lung cancer is the leading cause of cancer death and the second most commonly diagnosed cancer worldwide . Histolo1) genes ,4. EGFR nt NSCLC   or EGFR-independent mechanisms  [EGFR-mutant NSCLC with ARMs to osimertinib presenting multiple targetable alterations is still unclear. The molecular heterogeneity related to ARMs to EGFR TKIs emphasized the weakness of therapeutic options after progression on osimertinib. Platinum doublet chemotherapy-based regimens are recommended in patients who progress on osimertinib. Nevertheless, some EGFR-dependent ARMs, such as the EGFR C797S mutation, seem to be targetable with new fourth-generation EGFR TKIs in preclinical studies  or HER2 amplification and BRAF activation, are potentially targetable with third- or fourth-generation EGFR TKIs combined with targeted therapies against MET amplification, HER2 amplification, or BRAF activation. In this review, we discuss BRAF activation as an EGFR-independent ARM to osimertinib. We also tackle how we can use EGFR/BRAF/MEK co-inhibition as a new treatment strategy in BRAF activation as an ARM to osimertinib.Unfortunately, despite these remarkable outcome improvements in reatment . Tumor prmation) . The man BLU-945  and EAI0 BLU-945 ) and a cnt NSCLC . InteresBRAF is a member of the RAS/mitogen-activated protein kinase (MAPK) signaling pathway that mediates cell growth and malignant transformation [BRAF mutation is the highest in melanoma, observed in approximately one half of the cases [BRAF mutations observed in 2–6% of advanced-stage LUADs [EGFR mutations in NSCLC, which occur mainly in never-smokers, BRAF mutations are more frequent in former or current smokers [BRAF mutagenesis is also supported by the fact that the spectrum of BRAF mutations is limited to transversion mutations [BRAF-mutant LUADs naïve from any treatment do not harbor EGFR or KRAS co-mutations; BRAF V600, EGFR and KRAS mutations are mutually exclusive in this setting. However, BRAF-mutant LUADs can occur concomitantly to other genomic alterations, such as Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Alpha (PIK3CA) mutations or ALK translocations [BRAF mutation is the valine (V) to glutamate (E) substitution at residue 600 (V600E) located in exon 15 of the gene, which constitutively maintains BRAF in an active state [BRAF, the constitutively active BRAF V600 mutation does not require protein homodimerization to switch toward the active state [BRAF V600 mutation accounts for 50% of all BRAF mutations in advanced-stage lung cancer at diagnosis , epithelial-to-mesenchymal transition (EMT) through AXL receptor tyrosine kinase (AXL) activation, SCLC transformation and downstream pathway activation, such as PI3K mutations, and RAS–MAPK pathway activation, including BRAF mutations and rearrangements [BRAF activation leads, in tumor cells, to increased cell survival and proliferation, while apoptosis is decreased [In vitro and in vivo data showed that the RAS–MAPK pathway activation, through an GFR TKIs . Activatmutation   and praja ring finger ubiquitin ligase 2 gene (PJA2)/BRAF fusion partners have been described in BRAF rearrangements as an ARM to osimertinib. Activating BRAF rearrangements results in loss of the N-terminal auto-inhibitory domain of BRAF, leading to the formation of constitutively active BRAF fusion protein dimers that activate the RAS–MAPK pathway [ngements  have beengements , the mosmertinib , both in pathway ,27.BRAF genetic alteration as an ARM to osimertinib in EGFR-mutant NSCLC usually receive platinum-based chemotherapy. As of today, there is no therapeutic strategy using targeted therapy to inhibit both the EGFR and BRAF/MEK pathways. The management of tumors presenting multiple targetable mutations is still unclear. However, EGFR-independent ARMs include bypass signaling activation, such as MET and HER2 amplifications, which are targetable. For instance, in EGFR-mutant NSCLC harboring MET amplification as ARM to EGFR TKI, dual inhibition of EGFR and MET with the combination of osimertinib and savolitinib, respectively, provided promising results with an acceptable risk-benefit profile and encouraging antitumor activity [EGFR-mutant NSCLC patients with resistance to osimertinib in a phase 1 trial, CHRYSALIS [Patients experiencing tumor progression with activity . This EGactivity . Understactivity . The comHRYSALIS . UnfortuEGFR-mutant NSCLC in the first- or second-line setting  [EGFR T790M as well as the C797S mutations of resistance to first/second- and third-generation EGFR TKIs, respectively, and demonstrated interesting antitumor activity in preclinical studies [BRAF V600E mutation based on a phase 2 trial which showed an objective response rate (ORR) of 64%, a median duration of response of 15.2 months and a median PFS of 14.6 months, if assessed by the independent review committee, or 10.9 months, if investigator-assessed [Osimertinib is SOC in advanced-stage equired) ,7,35. Se studies . The cliassessed    , or vemuBRAF mutations, such as BRAF G469A, are other BRAF activation mechanisms which confer acquired resistance to osimertinib through the RAS–MAPK pathway activation. Importantly, in vitro data showed that the combination of osimertinib with a MEK inhibitor (selumetinib or trametinib) induced a synergistic effect, resulting in the suppression of proliferative pathways and the induction of apoptotic signaling, thus overcoming osimertinib resistance [Non-V600 sistance . These psistance ,38,39.BRAF activation through BRAF rearrangement is another ARM to osimertinib in EGFR-mutant NSCLC [BRAF rearrangement and an EGFR mutation in a synergistic manner  [EGFR-mutant NSCLC with MET amplification, while the combination of osimertinib and necitumumab is expected to be effective in EGFR-mutant NSCLC with EGFR amplification. Osimertinib plus selpercatinib and osimertinib plus alectinib could potentially be effective in EFGR-mutant NSCLC with RET and ALK rearrangements, respectively. Unfortunately, the targeted therapy approach against mechanisms leading to BRAF activation, which include V600 and non-V600 BRAF mutations and BRAF translocations which account for approximatively 5% of ARMs to osimertinib, has not been explored in clinical trials yet.The major clinical obstacle that limits the long-term benefit of osimertinib in patients with  of ARMs ,8,28. Thnt NSCLC ,40,41. T3778229)  and the 3778229) , is alsoEGFR-mutant NSCLC seems to be exclusively reported in patients treated with osimertinib [EGFR mutations. The proportion of BRAF activation may potentially increase in the context of tumor progressing on osimertinib in the adjuvant setting [EGFR-mutant NSCLC and BRAF activation as an ARM to EGFR TKIs [ALK-rearranged NSCLC. To do so, plasma genotyping needs to be considered during the treatment, as well as tumor biopsy at progression to detect the ARM to the EGFR/BRAF/MEK pathway co-inhibition. Interestingly, an ARM to EGFR/BRAF/MEK pathway co-inhibition, with osimertinib as the EGFR TKI, may be the EGFR C797S mutation, which is potentially targetable with the fourth generation EGFR TKI BLU-945 [This BRAF activation in mertinib , which h setting . FurtherGFR TKIs ,29,30,42 BLU-945 ,25,43.


Stress granules (SGs) are highly dynamic cytoplasmic foci formed in response to stress. The formation of SGs is reported to be regulated by diverse post-translational protein modifications (PTMs). Among them, ADP-ribosylation is of emerging interest due to its recently identified roles in SG organization. In this review, we summarized the latest advances on the roles of poly(ADP-ribose) (PAR) in the regulation of SG formation and dynamics, including its function in modulating nucleocytoplasmic trafficking and SG recruitment of SG components, as well as its effects on protein phase separation behavior. Moreover, the functional role of PAR chain diversity on dynamic of SG composition is also introduced. Potential future developments on investigating global ADP-ribosylation networks, individual roles of different PARPs, and interactions between ADP-ribosylation and other PTMs in SGs are also discussed. Stress granules (SGs) are cytoplasmic membraneless structures that rapidly assemble in cells in response to a variety of stresses, including heat, oxidative stress, and virus infection . These liso-ADP-ribose  of alphaviruses can remove the PARylation modification from the SG component G3BP1 and thus inhibit SG formation. This ability depends on the conserved macrodomain of nsP3 which function as a mono-ADP-ribosylhydrolase. Therefore, it is more likely that the nsP3 macrodomain prevents MARylation and subsequent indirectly PARylation, instead of PAR chain degradation . The rolHow does PAR regulate these SG related-processes? Recent in-depth studies have shown that PAR controls SG formation and dynamics via different mechanisms . The effDrosophila reduces TDP-43 cytoplasmic location, and in mammalian cells and neurons, PAR- or PAR-scaffold-binding through PBM is required for TDP-43 accumulation in cytoplasmic SGs , TAR DNA binding protein 43 kDa (TDP-43), Ras-GTPase activating protein SH3 domain-binding protein (G3BP), argonaute family member Ago2, and T-cell intracellular antigen-1 (TIA-1) . For exasmic SGs . hnRNP Asmic SGs . It can smic SGs . These ssmic SGs .in vitro or in vivo. For instance, a recent publication on apoptosis signal-regulating kinase 3 (ASK3) indicated that PAR could keep ASK3 condensates in the liquid phase and enable cells to sense osmotic stress (in vitro addition of PAR polymers promotes hnRNP A1 LLPS (in vitro assays have shown that PAR addition enhances co-LLPS of these two proteins (It has been reported that SG formation is induced by liquid-liquid phase separation (LLPS) of intrinsically disordered proteins (IDPs) and RNAs . IDPs lac stress . LLPS ofc stress . For ins A1 LLPS . And theproteins . Moreoveproteins . Thus, aproteins .It has been observed that PAR levels can affect protein phase separation behavior in vitro phase separation system, the dynamics of the hnRNP A1 droplet are reduced , and myopathy. This has been correlated with the aggregation-prone propensities of these mutants in the cytoplasm, resulting in SGs nucleation and pathological SGs formation . Althoug reduced . Moreove reduced . Other g reduced . The abo reduced . Therefo reduced . Hyperac reduced . Under l reduced . This is reduced .Taken together, PAR can serve as a scaffold for inducing LLPS of macromolecules and SG assembly. However, when PARP activity and PAR levels are mis-regulated, the proteins and granules may condense into irreversible solid phase aggregations that contribute to neuropathies .The PAR chain can vary from a few to hundreds of ADP-ribose units . StudiesPoly(ADP-ribose) has become an emerging research topic due to its recently identified roles in the organization of SGs and pathogenesis of SG-related neurodegenerative diseases . We summFurther in-depth investigations are needed to clarify the interplay between PAR and SGs under different conditions. Such as what are the PAR-mediated interaction networks in SGs. This includes systematically determination of SG components that are PARylated or PAR-bound under different stresses. Different groups have used diverse approaches to identify PARylated vs. PAR-binding proteins , which cThe other remaining question is what are the individual roles played by various PARPs in SGs, and whether or not they affect each other. Six reported SG-localized PARPs might have their own specific targets and functions. They might specifically regulate the localization, LLPS behavior, and interaction network of their substrates. For example, previous study indicated that long-term stress result in excessive cytoplasmic accumulation of the disease-related protein TDP-43, and reduction of PARP5a/5b activity could antagonize cytoplasmic aggregation of TDP-43 and its mediated neurotoxicity . TherefoMoreover, what are the links between ADP-ribosylation and other PTMs. Do they work together synergistically or compete with each other on substrates via conjunction on the same or adjacent motifs in SG regulation? A previous study indicated that PAR-dependent anchoring of TDP-43 to SGs inhibits its disease-associated phosphorylation. If TDP-43 were to accumulate excessively in the cytoplasm and become excluded from SGs, it could become phosphorylated and form irreversible aggregates . TherefoPoly(ADP-ribose) as an important novel regulator of SG formation and dynamics, in-depth investigations on the remaining questions discussed above will help us to acquire a clear mechanistic view on global PAR-mediated interaction networks, specific functions of PARPs and links between ADP-ribosylation and other PTMs in SGs. This will eventually contribute on the development of novel therapeutic approaches targeting the aberrant pathogenic SG formation and further pave the way for effective SG-related neurodegenerative disease treatments.XJ, XC, and BL contributed to the design and wrote the manuscript. BL and SL supervised the overall direction, planning, and editing of the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


KRAS (encoding the KRAS proto-oncogene) and remain difficult to target in the clinic. We have recently shown that activation of KRAS protein alters the regulation of mRNA translation, increasing total protein synthesis, and maintaining elevated c-MYC (MYC proto-oncogene) expression. Targeting these pathways downstream of KRAS reveals a striking dependency that has potential for clinical translation.~50% of colorectal cancers have an activating mutation in  KRAS-mutant patient-derived organoids to investigate how KRAS activation alters the fundamental cellular process of protein synthesis.1 We found that activation of KRAS nearly doubled the rate of protein synthesis in organoids deficient for the tumor suppressor APC   with rapamycin greatly suppressed intestinal adenoma proliferation, 2 and is in agreement with the drug resistance found in KRAS-mutant tumors in the clinic. An independent study in organoids of the same genotype as our mouse models found that Kras mutation alone has no effect on protein synthesis rates, but in conjunction with Apc-deficiency does lead to an increase.3 Interestingly, this same paper found further increases in protein synthesis upon suppression of the commonly mutated genes, Smad4 (SMAD family member 4) and Trp53 (transformation-related protein 53). This highlights a direct link between tumor mutations and protein synthesis that, when fully understood, could provide methods for personalized therapeutic intervention.Mutations that result in the constitutive activation of KRAS signaling  are present in nearly half of all colorectal cancers (CRCs), with these patients responding poorly to current treatments. The design of targeted therapies is reliant upon understanding how mutant KRAS alters tumor cells. We recently used genetically engineered mouse models and pathway) . FurtherApc-deficient, Kras-mutant mouse models, despite not affecting proliferation. From this, we draw two conclusions. First, that a proportion of translation occurring in Kras-mutant cells is not required for proliferation and, secondly, there is not a linear relationship between protein synthesis and proliferation rates. In agreement with this, targeting signaling to increase the rate of protein synthesis in CRC models via eIF2 (eukaryotic translation initiation factor 2) is effective at reducing proliferation and inducing apoptosis.4 In this instance increasing protein synthesis suppressed proliferation, highlighting the presence of a protein synthesis window that is permissive to cancer-associated hyper-proliferation , the mRNA cap-binding protein. This phosphorylation, performed by either MNK1 or MNK2 (MAPK interacting serine/threonine kinases 1 or 2), is believed to enhance the translation of a subset of mRNAs, many of which are implicated in tumorigenesis [reviewed by P-eIF4E) we found expression of the proto-oncogene c-MYC is maintained by this signaling downstream of KRAS. Importantly, additional targeting of protein synthesis was required to reduce c-MYC levels, which correlated with suppression of proliferation. Specifically, a combination of rapamycin to suppress protein synthesis, and the MNK inhibitor eFT508 to suppress P-eIF4E-dependent translation, produced a dramatic reduction in proliferation and a more than 10-fold extension of survival in mice bearing Apc-deficient Kras-mutant intestinal adenomas. Supplementing our pre-clinical results, we analyzed clinical CRC samples and identified a positive correlation between mTORC1 and MNK activity, with nearly half of patients having elevated signaling through both pathways. Furthermore, stratification found a subgroup of these patients that had a 3.5-year shorter cancer-specific survival compared to those with low signaling through either or both pathways. This group accounted for 20% of patients studied.Our work found that the expression of mutant KRAS-mutant CRC. We found similar effects when combining eFT508 with other compounds that suppress protein synthesis, such as oxaliplatin and 5-fluorouracil. eFT508 is currently being trialed in combination with paclitaxel in advanced breast cancer (NCT04261218) with paclitaxel known to suppress protein synthesis in cultured cells.6 Interestingly, genetic or chemical inhibition of P-eIF4E reverses chemotherapy-induced neuropathic pain induced by paclitaxel treatment in vivo, 7 indicating that there may be additional benefits of clinical targeting of P-eIF4E.eFT508  is an exceptional, well-tolerated compound that is progressing through eight clinical trials against multiple solid tumors. Our work provides pre-clinical validation for the combination treatment of eFT508 with rapalogues in c-MYC mRNA was significantly enriched in P-eIF4E immunoprecipitations from mouse organoids and human cell lines, identifying c-MYC as an essential effector of P-eIF4E. This is supported by our observation that increasing c-MYC expression in vivo was sufficient to completely reverse the benefit of our targeting strategy. Another recent publication found that the association of c-MYC mRNA with polysomes, the translation hubs of the cell, was significantly reduced in the human colon carcinoma cell line HCT116, where one copy of eIF4E could not be phosphorylated.8 This correlated with reduced c-MYC protein levels and cell growth. HCT116 cells harbor a mutation in KRAS (G13D) and activated Wnt-signaling via mutation of β-catenin (CTNNB1), making them similar to the Apc-deficient (Wnt activated), Kras-mutant mouse models used in our publication. Thus, the synthesis of c-MYC is highly dependent on P-eIF4E in two systems modeling CRC. Furthermore, the translation of c-MYC is undoubtedly important in a range of other tumor settings, making this targeting strategy potentially applicable to other MYC-dependent malignancies.Mechanistically, we showed that 9 During the search for targeted therapies that rely on biological understanding, the regulation of translation will be particularly amenable to therapeutic intervention. Targeting P-eIF4E provides an example of this, but clinical trials are currently underway using compounds targeting other parts of the translation machinery, providing clinicians with the tools to target translation initiation in cancer.There is growing interest in targeting protein synthesis to suppress tumors. Many first-line chemotherapies suppress translation, indicating that this strategy has, in fact, been used for decades.KRAS mutant colorectal cancers. Targeted therapies are also being developed to increase protein synthesis rates in tumors, thereby overloading proteotoxic stress response pathways and reducing tumor proliferation. This strategy may be particularly effective against tumors with multiple mutations, where the permissive rate of protein synthesis may nearly be exceeded.The rate of protein synthesis is increased by cancer-associated mutations in colorectal cancer, such as APC  loss or KRAS (KRAS proto-oncogene) mutation. Increased protein synthesis is permissive for hyperproliferation, with lower levels unable to support increased proliferation. However, it is also possible for excessive protein synthesis to occur, resulting in diminished proliferation and apoptosis. Tumor cells, therefore, tightly regulate their translation rate to remain in the permissive region. Therapies targeting protein synthesis can either decrease or increase overall rates. Many first-line chemotherapies suppress ribosome synthesis, and in turn translation, contributing to their efficacy as anticancer agents. These include 5-fluorouracil (5-FU) and oxaliplatin. More recently, targeted therapies against specific kinases regulating the translation machinery or translation factors themselves have been shown to suppress protein synthesis in pre-clinical models and have some efficacy in the clinic. Our recent article demonstrated the ability of targeting eIF4E (eukaryotic initiation factor 4E) phosphorylation to restore sensitivity to agents suppressing translation in


Lymphovascular space invasion (LVSI) is the first step of hematogenous metastasis. Exploration of the differential miRNA expression profiles between LVSI-positive and LVSI-negative ovarian cancer tissues may help to identify key miRNAs involved in the hematogenous metastasis of ovarian cancer. This study is aimed to identify microRNAs (miRNAs) that are differentially expressed between LVSI-positive and LVSI-negative ovarian cancer tissues, followed by exploring their association with bevacizumab response in ovarian cancer patients.The Cancer Genome Altas (TGGA) dataset was used to identify the differentially expressed miRNAs between LVSI-positive and LVSI-negative ovarian cancer tissues. The prognostic value of the differentially expressed miRNAs was determined using GSE140082 dataset.We showed that miR-25 and miR-142 were differentially expressed between LVSI-positive and LVSI-negative ovarian cancer tumors. Kaplan-Meier analysis indicated that high miR-25 expression was associated with increased progression free survival (PFS) and extended overall survival (OS). Moreover, patients with low miR-25 expression benefited significantly from bevacizumab treatment in terms of PFS. A similar trend was observed in terms of OS though without reaching statistical significance. In contrast, no significant survival benefits from bevacizumab were observed in patients with high miR-25 expression in terms of PFS and OS. There was no significant correlation between miR-142 expression and PFS. In contrast, high miR-142 expression was associated with reduced OS. Moreover, patients with high miR-142 expression benefited significantly from bevacizumab treatment in terms of PFS and OS. However, bevacizumab treatment conferred no significant improvements in both PFS and OS in patients with low miR-142 expression. The nomogram for PFS indicated that miR-25 expression had a larger contribution to PFS than debulking status and bevacizumab treatment. And the nomogram for OS illustrated both miR-25 expression and miR-142 expression as sharing a larger contribution to OS than bevacizumab treatment and debulking status.In conclusion, miR-25 expression correlates with a better PFS and OS in ovarian cancer. Patients with low miR-25 expression and high miR-142 expression could benefit from bevacizumab treatment significantly. Ovarian cancer is the most lethal gynecological malignancy, and most of the cases are not diagnosed until advance stages . PatientFor many years, ovarian cancer has been widely believed to spread via a passive mechanism . Namely,In this study, we tried to identify miRNAs that were differentially expressed between LVSI-positive and LVSI-negative ovarian cancer tissues, and subsequently explored the association of LVSI-related miRNAs with bevacizumab response in ovarian cancer patients.P value< 0.05 were considered to be statistically significant.LVSI information and miRNA profiling were available in 191 ovarian cancer patients in the TCGA dataset. For these patients, we downloaded the level 3 miRNA expression data. Differential expressed genes between LVSI-positive and LVSI-negative tumors are identified using the R package Limma . Fold chGSE140082 dataset was used to explore the prognostic value of LVSI-associated miRNA in ovarian cancer. Namely, univariate and multivariate analysis were used to explore the association of miR-25 and miR-142 with progression free survival (PFS) and overall survival (OS) in patients with ovarian cancer. Bevacizumab response were also evaluated according to the expression level of miR-25 and miR-142.P value < 0.05 was considered statistically significant.Limma R package was used to identify the differentially expressed miRNAs between LVSI-positive and LVSI-negative ovarian cancer tissues. “Low” and “high” was defined according to the gene expression level. The cutoff values for miR-25 and miR-142 were determined using survminer R package. Continuous data between two groups and categorical data were compared using two independent samples t-test and chi-square or Fisher’s exact test where appropriate, respectively. Univariable and multivariable analyses were used to identify factors associated with PFS and OS. Details of the development of the prognostic nomograms were described in our previous published paper . The staP value< 0.05, respectively  and upregulated (fold change> 1.25) in LVSI-positive group versus LVSI-negative group with a P = 0.005) and extended OS . There was no significant correlation between miR-142 expression and PFS . In contrast, high miR-142 expression was associated with reduced OS .Then, we explored the associations of miR-25 and miR-142 expression with the survival of ovarian cancer patients. Kaplan Meier analysis indicated that high miR-25 expression was associated with increased PFS . A similar trend was observed in terms of OS  though without reaching statistical significance. In contrast, no significant survival benefits from bevacizumab were observed in patients with high miR-25 expression in terms of PFS  and OS . And patients with high miR-142 expression benefited significantly from bevacizumab treatment in terms of PFS  and OS . However, bevacizumab treatment conferred no significant improvements in both PFS  and OS  in patients with low miR-142 expression.Since LVSI is closely correlated with hematogenous metastasis of cancer cells, we infer that the abundance of LVSI-associated miRNAs may be predictive of response to anti-angiogenesis therapy in ovarian cancer. Thus, we next explored whether molecular subtyping by miR-25 and miR-142 expression analysis could identify patients who would preferentially benefit from bevacizumab treatment. Our results indicated that patients with low miR-25 expression benefited significantly from bevacizumab treatment in terms of PFS  gene. The expression of miR-25 was tissue specific. Previous studies showed that miR-25 was upregulated in various cancers, such as breast cancer, endometrial cancer, and so on. However, in prostate cancer and colorectal cancer, the results were contradictory. Some studies showed miR-25 was upregulated in prostate cancer and colorectal cancer. Other studies, however, showed miR-25 was downregulated in prostate cancer and colorectal cancer. In ovarian cancer, Wang et al. showed that miR-25 was upregulated and its expression was positively associated with tumor stage and regional lymph node involvement . Namely,MiR-142, first discovered in chromosome 17 of hematopoietic stem cell, has been demonstrated to be a critical regulator of various biological processes, and dysregulation of miR-142 was associated with various diseases, including cancer. MiR-142 could function as either tumor suppressor or oncogene in a cell-specific manner. For example, miR-142 overexpression could regulate both immune response and tumor microenvironment to initiate a powerful anti-tumor response . In contConsidering the survival benefit from adding bevacizumab to the frontline therapy in GOG-218 trial and ICON7 trial, bevacizumab was approved as the upfront therapy for ovarian cancer , 18. HowIn conclusion, miR-25 expression correlates with a better PFS and OS in ovarian cancer. Patients with low miR-25 expression and high miR-142 expression could benefit from bevacizumab treatment. Further researches are needed to explore the mechanism on the relationship between bevacizumab response and the expression of miR-25 and miR-142.


To achieve this, root cells integrate signals from the local external availability of nutrients with their own internal signals to modulate the activity of root uptake systems, thereby matching the nutrient needs of the whole plant. Nitrogen (N) is an important macronutrient modifying RSA, and nitrate (arvation .). Diverse and delicate molecular events underlie these changes. The primary nitrate response is marked by massive and rapid transcriptional and post-transcriptional regulation of N transporters, assimilation enzymes, and signaling factors . Depending on ;Jia et al., 2020). While molecular, physiological, and transcriptomic observations demonstrate the nitrate regulation of auxin transport and biosynthesis to stimulate lateral root emergence and lateral and primary root elongation to low-nitrate conditions, such observations for BRs are still emerging.RSA modification in response to nitrate deficiency includes morphological changes in lateral and primary roots, such as promotion of lateral and primary root elongation and lateral root number . Diverse factors . DependiPlant Physiology, Arabidopsis thaliana) as a model plant. They report this module links low-nitrate and BR signals for coordinating root development to prevent quick depletion of N resources. In a previous study, the authors identified diverse root morphology under different nitrate levels and between natural accessions of Arabidopsis (CML38 as a causative gene responsible for such variation and set out to characterize its protein function in Columbia (Col-0). RT-qPCR and promoter-β-glucuronidase assays confirmed the CML38. Mutation of CML38 increased primary root length in low- and high-nitrate conditions, and lateral root number in low-nitrate conditions. The authors investigated the role of CML38 in N uptake, assimilation, and metabolism through biochemical assays and showed increased absorption and assimilation of cml38, an observation consistent with the upregulation of primary nitrate-response genes, including those encoding various N-transporters and -reductases. They conclude that CML38 participates in the primary nitrate response in Arabidopsis.In this issue of bidopsis . In thei. However, PEPR2 function in N-signaling had not been investigated previously. Through double and triple mutant analyses, the authors confirmed that PEPR2 and CML32 likely work in the same genetic pathway to co-regulate . However), and mild N deficiency upregulates the transcript levels of BAK1 to activate BR signaling and stimulate root elongation . Hence, CML38 and PEPR2 in roots. Mutation of CML32, PEPR2, or both increased primary root length and lateral root number upon BR supply, suggestive of their positive regulation in BR signaling. Combining either low- or high-nitrate conditions with BRs, the authors further confirmed that low-cml32 and pepr2 mutant roots exposed to BR plus low-The authors further confirmed that CML38 and PEPR2 play crucial roles in the BR signaling pathway promoted by low nitrate. PEPR2 interacts with BRI1-ASSOCIATED KINASE1 (BAK1), a master regulator of BR signaling , and milongation . Hence, These results demonstrate CML38 and PEPR2 as important molecular players in the local N signaling pathway that holds many intertwined connections to the root developmental processes and BR signaling. It remains to be studied further how different common target genes of CML38-PEPR2 mechanistically coordinate such connections and whether CML38 and PEPR2 may act in combination to mediate a root-to-shoot-to-root-relay of systemic N-demand signaling . Understanding such hierarchical regulation may allow biotechnological engineering of N-efficient crops and improve yield.Conflict of interest statement. None declared.


Apolipoprotein (Apo)-based mimetic peptides have been shown to reduce atherosclerosis. Most of the ApoC-II and ApoE mimetics exert anti-atherosclerotic effects by improving lipid profile. ApoC-II mimetics reverse hypertriglyceridemia and ApoE-based peptides such as Ac-hE18A-NH2 reduce cholesterol and triglyceride (TG) levels in humans. Conversely, other classes of ApoE and ApoA-I mimetic peptides and, more recently, ApoJ and LRP1-based peptides, exhibit several anti-atherosclerotic actions in experimental models without influencing lipoprotein profile. These other mimetic peptides display at least one atheroprotective mechanism such as providing LDL stability against mechanical modification or conferring protection against the action of lipolytic enzymes inducing LDL aggregation in the arterial intima. Other anti-atherosclerotic effects exerted by these peptides also include protection against foam cell formation and inflammation, and induction of reverse cholesterol transport. Although the underlying mechanisms of action are still poorly described, the recent findings suggest that these mimetics could confer atheroprotection by favorably influencing lipoprotein function rather than lipoprotein levels. Despite the promising results obtained with peptide mimetics, the assessment of their stability, atheroprotective efficacy and tissue targeted delivery are issues currently under progress.  In the past decades, worsening of eating habits and sedentary lifestyle factors have increased worldwide incidence of pathologies associated with cardiovascular risk. Nowadays, cardiovascular diseases cause more than 17.5 million deaths annually, representing an enormous health and socioeconomic problem. Moreover, population aging and gradual accumulation of risk factors  are key determinants of the growth of cardiovascular disease prevalence. One of the main underlying causes of cardiovascular diseases is atherosclerosis. Atherosclerotic lesions are strongly and uninterruptedly promoted by cholesteryl ester-enriched lipoproteins, mainly lipoprotein remnants and low-density lipoproteins (LDL). Modified lipoproteins accumulate in the arterial wall and exert locally proatherogenic actions, including the induction of intracellular lipid accumulation and inflammation ,3.There are several therapeutic strategies that combine healthy lifestyle with pharmacological interventions to modulate atherosclerosis through the control of lipoprotein metabolism. The most widely used drugs in primary and secondary prevention are statins or HMG-CoA reductase inhibitors. The inhibition of HMG-CoA reductase reduces intracellular cholesterol and increases LDL receptor levels in the liver, facilitating increased blood LDL clearance . DespiteStatin treatment was not always associated to a reduction in cardiovascular mortality . Other ePeptides and their mimetics with capacity to improve lipid profile and/or lipoprotein functionality have emerged as potential therapeutic tools in cardiometabolic diseases and particularly, in atherosclerosis. These peptides are usually designed as short amino acid chains based on functional domains of apoproteins or apoprotein receptors. Advanced technologies have helped to progressively improve the therapeutic characteristics of these peptides. There are several peptides that have already met a wide range of therapeutic objectives, including good affinity profiles, oral bioavailability, low toxicity, biosecurity, tolerability, good efficacy, high potency, and selectivity .Recombinant human insulin (Humulin) was the first peptide produced for cardiometabolic therapy , in partIn the vascular field, calcitonin gene-related peptide (CGRP) is a neuropeptide produced by calcitonin gene alternative RNA processing. CGRP is a potent vasodilator that protects the cardiovascular system and facilitates wound healing .According to Transparency Market Research (TMR), the market value of cardiovascular drugs will exceed US$ 91 billion by 2025 . The bulIn this review, we focus on mimetic peptides with anti-atherosclerotic properties, highlighting peptide-preventing effects on LDL instability and aggregation. LDL aggregation is a key determinant of LDL proatherogenicity. This review (1) updates in vitro and in vivo studies using mimetic peptides and their influence on atherosclerosis, (2) discusses the therapeutic potential of LRP1-derived peptides, and (3) summarizes clinical trials using apolipoprotein-based peptides.Apolipoprotein and LRP1-based peptides are short, chemically synthesized peptides that mimic amino acid sequences of natural apolipoproteins or lipoprotein receptors. Given their amphipathic nature, these peptides bind to lipoproteins, conferring structural and functional protection . UsuallyApolipoprotein A-I (ApoA-I), the main apolipoprotein in HDL, is essential for HDL generation and function. ApoA-I is composed of 243 amino acids structured in 10 amphipathic α-helices, most of them essential for lipid interchange . The intPeptides derived from ApoA-I, and particularly peptide 4F, have been shown to exert not only anti-LDL-aggregating effects, but an additional wide variety of anti-atherosclerotic effects ,39. 4F pApolipoprotein C-II (ApoC-II) is an exchangeable protein composed of three amphipathic α-helices that are located at the surface of chylomicrons, HDL, and very-low-density lipoproteins (VLDL) lipoproteins. In the postprandial state, ApoC-II is mainly carried by VLDL and LDL . ApoC-IIApolipoprotein E (ApoE) is a multifunctional secreted glycoprotein of 299 amino acids mainly synthesized by the liver. ApoE consists of two domains separated by a hinge region: the LDLR-binding globular domain (1–191 residues) and the lipid-binding domain with an amphipathic α-helical motif (192–299 residues) . The gloApoE genetic variations are associated not only with plasma lipid levels but also with atherosclerosis risk and pathobiology of neurodegenerative diseases ,47.2, a dual-domain cationic apolipoprotein-mimetic peptide composed of the LDLR-binding region of ApoE (141–150 residues), and the peptide 18A , also referred as clusterin is a 449 amino acid protein that acts extra- or intracellularly as a chaperone. The mature protein is a glycosylated heterodimer formed by α- and β-chains linked through five disulfide bonds. Approximately 20.5% of circulating ApoJ is associated with lipoproteins , although the content of lipoprotein-associated ApoJ decreases in hyperlipidemia . In HDL,Navab et al. synthesized seven peptides corresponding to seven G1 amphipathic helices of apoJ. Among them, the active peptides correspond to 113–122 residues, which were synthesized as D-amino acids . D-, LRP1 is the main receptor responsible for the uptake of esterified cholesterol from agLDL . Hyperchom agLDL . From thionality ,59.3 and D-Glu9) and two positively charged ApoB-100 residues (Lys3229 and Lys3234) [Biochemical studies demonstrated that both LP3 and DP3 peptides prevent SMase- and phospholipase A2 (PLA2)-induced LDL aggregation by binding to a specific sequence of ApoB-100 . LDL proA new family of LRP1-P3 peptide derivatives with enhanced potency and proteolytic stability were designed through in silico conformational sampling and ApoB-100 molecular docking. These peptides  were tested using a dual  screening assay. This new family of peptides contains linear, fragment, cyclic, and alanine scanning derivatives and has been generated through two consecutive optimization rounds. Structurally and functionally optimized peptides contain hotspot residues that were replaced by alanine, because this strategy confers an increased capacity to form prone α-helix conformations that are crucial for electrostatic interaction with ApoB-100. These new compounds were proven to efficiently inhibit LDL aggregation promoted by SMase and PLA2 .In this section, we review the mechanisms underlying the effects of these peptides in atherosclerosis.Both L-4F and D-4F peptides reduce atherosclerotic plaque burden, pro-inflammatory cytokine secretion, insulin resistance, and lipid oxidation in biochemical, cellular, and in vivo models ,61. A diThe 18A-C-II-a peptide enhances lipolysis in plasma from patients with ApoC-II deficiency and with other forms of hypertriglyceridemia in ex vivo studies . This pe2 reduces plasma cholesterol, promotes macrophage cholesterol efflux, shows anti-inflammatory and anti-oxidative properties, and reduces atherosclerotic lesion formation to a greater extent than 4F in ApoE-deficient mice [2 also decreases total cholesterol and lipid peroxide levels and increases HDL-associated PON activity in the plasma of Watanabe hypercholesterolemic rabbits [ApoE has protective effects on the progression of atherosclerosis by promoting the hepatic uptake of atherogenic lipoproteins and macrophage cholesterol efflux . Consistent mice . Ac-hE18 rabbits . A modif rabbits .Oral administration of mR18L, the cationic single domain amphipathic peptide, reduces cholesterol plasma levels and atherosclerotic lesion in ApoE-deficient mice . In LDLRThe EpK peptide, which contains an LDLR-binding region and a lipid-binding region, binds to HDL and protects macrophages against LPS-induced inflammation. This peptide also promotes cholesterol efflux with higher efficiency than ApoA-I and ApoE3 . In addiApoJ expression increases under stress and forms part of the control system against protein unfolding. ApoJ is involved in apoptosis, cell adhesion, tissue remodeling, immune system regulation, and oxidative stress . As a coSmooth muscle cells are the main cellular components of the vascular wall, and up to 40% of foam cells previously identified as monocyte-derived macrophages in human atherosclerosis are SMCs ,83. One Several studies have reported the efficacy of newly developed peptides  to induc−/− mice [The recognition of modified oxidized LDL (oxLDL) by the immune system is a key step in the activation and regulation of the inflammatory process that occurs during atherosclerosis . Anti-ox−/− mice . ImmunizApoB-100-based peptides show pro-inflammatory properties such as activation of T cells, B cells, and monocyte-macrophage system. Therefore, therapies based on immunization with these peptides have been developed .−/− mice [+ T-cell response activation [Immunization with p210 peptide (3136 to 3155 residues of human apoB-100) has been shown to decrease dendritic cell and macrophage infiltration, and reduce atherosclerotic plaque burden in ApoE−/− mice ,92. PeptAntibodies against LDL(-) have been detected in the blood and atherosclerotic plaques of rabbit models and humans ,100. TheLRP1 interacts with several anti-inflammatory ligands including ApoE that blocks interleukin-1 receptor associated kinase-1 (IRAK-1) activation, helping to reduce the inflammatory effects of nuclear factor-kappa B (NF-κB) in VSMCs . LRP1 coSeveral peptides are currently undergoing clinical evaluation. As shown in Among all ApoA-I-derived peptides, few have succeeded in clinical studies. The oral administration of D-4F reduced HDL inflammation index (HII) in high-risk cardiovascular patients . HII wasIn other clinical study, L-4F peptide was administered either via intravenous infusion over 7 days or via subcutaneous injection over 28 days to patients with CAD . Blood pThe peptide 5A combined with sphingomyelin, called Fx-5A, is currently undergoing a phase 1A clinical trial . This peSeveral molecules mimicking HDL/ApoA-I have been developed to enhance RCT. Two of these molecules, ApoA-I Milano and CSL-112, have been already tested in clinical trials. ApoA-I Milano peptide was administered intravenously preparing a peptid/lipid complex (ETC-216) and its efficacy was assessed through the evaluation of atherosclerotic plaque burden by means of intravascular ultrasound (IVUS) . This peAnother ApoA-I mimetic complexed with phospholipids, CER-001, was investigated in patients with a history of percutaneous coronary intervention (PCI) or with at least 20% coronary artery luminal narrowing. This phase 2 study revealed that CER-001 did not reduce coronary atherosclerosis, as assessed by IVUS and quantitative coronary angiography (QCA) . In anot2 peptide, under the commercial name AEM-28, demonstrated its safety profile in phase 1a and phase 1b/2a clinical trials. Intravenous administration to 51 patients resulted in a reduction of over 50% in TGs and VLDL-C compared to placebo treatment [The Ac-hE18A-NHreatment .The development of liver X receptor (LXR) agonists and microsomal triglyceride transfer protein (MTP1) drugs as emergent therapies for atherosclerosis were a big challenge in the last years. LXR are essential nuclear receptors that exert atheroprotective properties due to their capacity to decrease intestinal cholesterol absorption and induce RCT. Therefore, a big effort was invested to develop efficient LXR agonists . MTP is This is a comprehensive review focused on the use of peptides as anti-atherosclerotic tools. It integrates results from in vitro and in vivo studies, and clinical trials to summarize the potential applicability of these novel therapeutic tools. We hope to arise the interest of researchers and clinicians involved in the Atherosclerosis field. As shown in We performed an extensive (systematic) search in PubMed by combining several terms, mainly: mimetic peptides, apolipoprotein, LRP1, atherosclerosis, LDL, and HDL. Relevant articles were chosen and their references were secondarily searched for additional relevant articles with no limit on the original date of the article.