O-methylated epigallocatechin gallate (EGCG), resveratrol, curcumin, and cinnamon extract improved symptoms of allergic rhinitis (AR) and reduced the number of inflammatory cells in patients. However, strategies to overcome the poor bioavailability of these nutrients are an important part of current research.Allergic diseases affect an estimated 30 percent of the world’s population. Mast cells (MC) are the key effector cells of allergic reactions by releasing pro-inflammatory mediators such as histamine, lipid mediators, and cytokines/chemokines. Components of the daily diet, including certain fatty acids, amino acids, and vitamins, as well as secondary plant components, may have effects on MC and thus may be of interest as nutraceuticals for the prevention and treatment of allergies. This review summarizes the anti-inflammatory effects of dietary components on MC, including the signaling pathways involved, in in vitro and in vivo models. Butyrate, calcitriol, kaempferol, quercetin, luteolin, resveratrol, curcumin, and cinnamon extract were the most effective in suppressing the release of preformed and de novo synthesized mediators from MC or in animal models. In randomized controlled trials (RCT), vitamin D, quercetin,  It is estimated that more than 30% of the world’s population is affected by one or more allergic diseases. And the prevalence is increasing worldwide ,2. The mIn general, allergy is an overreaction of the immune system to a normally harmless substance, usually a protein, which manifests itself as a skin rash, sneezing, swelling of the mucous membranes, or other abnormal conditions . The terMC promote inflammation by releasing a wide range of inflammatory mediators upon activation. These include pre-stored mediators in MC granules, such as histamine and proteases, as well as de novo synthesized mediators such as cytokines and eicosanoids . The traMC play a key role in the pathogenesis of allergic diseases, including allergic rhinitis, asthma, atopic dermatitis, food allergy, drug allergy, and the life-threatening systemic MC-mediated reaction known as anaphylaxis ,17. For The effects of these dietary components on MC activation can be studied in vitro in rodent and human MC models and in vivo on rodent animal models of allergic diseases . There aBecause of the potential effects of these dietary components on MC in vitro and in vivo , they co2+ and Lyn-p38/NF-κB signaling pathways. Similarly, ALA exerted the same effects on serum histamine and cytokine release while reducing vasodilation and the percentage of degranulated MC in C57BL/6 mice [Fatty acids are crucial components of lipid membranes and can influence the production and secretion of mediators by MC . In partL/6 mice .EPA and DHA, which can be synthesized in the body from ALA, reduced the release of β-hex (≈25%) and cysteinyl-leukotriene (cys-LT) (≈80%) by LAD2 cells . EPA and2, IL-4, and IL-13 by more than 60%. The inhibition of IL-4 and IL-13 secretion was hereby correlated with a reduced generation of reactive oxygen species and suppression of MAPK. On the other hand, the omega-6 fatty acid arachidonic acid (AA) enhanced the production of proinflammatory MC mediators, including increased secretion of PGD2 and TNF-α by HMC-1. These findings suggest that EPA and DHA as omega-3 fatty acids may have beneficial effects in modulating MC function and reducing inflammation, whereas omega-6 fatty acids such as arachidonic acid may have pro-inflammatory effects [EPA and DHA may bind to and activate GPR120 in MC, potentially leading to increased cyclic adenosine monophosphate levels and the activation of protein kinase A, which could suppress MC degranulation . Further effects .MC in the gut and vascularized tissues are exposed to high concentrations of SCFA . SCFA buOral supplementation of sodium butyrate to weaned piglets reduced histamine release in the jejunal mucosa by about 40%. Gene expression and release of proinflammatory cytokines, including IL-6, TNF-α, and IL-13, decreased. The phosphorylation ratio of the MAPK JNK showed a decrease of about 60% after sodium butyrate feeding. However, no significant changes in the phosphorylation of ERK and p38 were observed. These results suggest that sodium butyrate improves the intestinal barrier function in weaned pigs by inhibiting the JNK signaling pathway . This su4 (LTC4) secretion by about 45%, but not β-hex release. High-dose treatment with both amino acids decreased chemokine expression of CC chemokine ligand (CCL)-2, CCL4, CXCL8, and TNF-α up to 50% in hiMC. Arginine and glutamine acted by suppressing the phosphorylation of the MAPK ERK, p38, and JNK, as well as of protein kinase B (Akt). Since the release and expression of the aforementioned MC mediators is regulated in part by the activation of MAPK and Akt, arginine and glutamine may modulate the inflammatory responses of MC [2 (≈70%) in response to dietary fat. This suggests that L-glutamine, but not D-glutamine, may accelerate proinflammatory responses leading to mucosal MC activation [The amino acids glutamine, arginine, and glycine have immunomodulatory and anti-inflammatory properties ,40. We ses of MC . Importativation . The amino acid glycine reduced the secretion of the proinflammatory cytokines TNF-α, IL-4, and IL-13, as detected in RBL-2H3 cells after IgE-mediated stimulation. Notably, no cellular degranulation was observed . Oral ad3) reduced IgE-mediated histamine release, cys-LT, and TNF-α by about 30% in BMMC in a VDR-dependent manner. The treatment of CBMC or PBMCMC with calcitriol or calcidiol resulted in a modestly reduced or comparable effect on the secretion of histamine, cys-LT, and TNF-α as in BMMC. MC 25-hydroxyvitamin D-1α-hydroxylase (CYP27B1) converted the intermediate metabolite calcidiol into calcitriol, which further suppressed MC activation. Calcidiol, although inactive, could bind to VDR with low affinity, but it is unclear whether this direct binding has suppressive effects on MC activation [Vitamin D is a fat-soluble vitamin with physiological and immunomodulatory properties ,99. It iylase CYPB1 convertivation . HIF-1α tivation . VEGF istivation . The appylase CYPB1 converCarotenoids are a broad group of natural pigments with a tetraterpene structure that possess anti-oxidant and anti-inflammatory properties . Rich so2 (PLA2), cyclooxygenase-2 (COX-2), and hyaluronidase in vitro [Oral fucoxanthin reduced mouse ear swelling induced by arachidonic acid, PMA, or oxazolone (OXA) by 20–28%, while percutaneous administration reduced it by more than 50% . Both pein vitro . Astaxanthin administration in PMA and calcium ionophore A23187 stimulated RBL-2H3 cells reduced the release of histamine and β-hex by about 30% . In a muThe carotene lycopene reduced the cytokine expression of IL-4, IL-13, and IL-9 by about 60% in an ovalbumin (OVA)-induced food allergy model . IL-9 ha2 release by about 40% in LAD2 cells [2+ downstream signaling [Flavonoids are a group of naturally-occurring polyphenolic plant substances with antioxidant and anti-inflammatory properties ,111,112.D2 cells . Kaempfeignaling ,46,49,50ignaling . Quercet2 release almost completely [In RBL-2H3 cells, Aceriphyllum rossii (ARE), a substance rich in quercetin and kaempferol, inhibited the histamine release by about 40% and PGEmpletely . ARE alsmpletely , but redmpletely . Kaempfempletely . A highempletely . IgE/antmpletely ,53. IL-3mpletely . Luteolimpletely , reducedmpletely . IL-33-smpletely . Epigallocatechin gallate (EGCG), a tea catechin, reduced both histamine degranulation and cytokine release  in receptor activator of NF-κB ligand (RANKL)-stimulated HMC-1 cells by targeting PI3K, MAPK, caspase-1, and NF-κB signaling cascades . Oral EGO-(3-O-methyl) gallate (EGCG″Me), an O-methylated catechin and found in the Benifuuki green tea, showed no significant effect on MC degranulation in IgE/antigen-activated RBL-2H3 cells but did when the bioactive flavanone eriodictyol or the citrus flavanone hesperetin was added. Combined administration of EGCG3″Me with eriodictyol or hesperetin inhibited IgE-mediated degranulation by inducing 67-kDa laminin receptor/soluble guanylate cyclase acid/acid sphingomyelinase signaling and enhancing this pathway. In a PCA mouse model both eriodictyol and highly absorbable α-glucosyl hesperidin potentiated the anti-allergic effect of Benifuuki green tea, with eriodictyol having a stronger effect on PCA responses (≈50%) [(-)-Epigallocatechin-3-s (≈50%) ,59.+ T cells, CD8+ T cells, degranulated MC (≈40%), suppression of serum IgE levels, and a reduced production of IFN-γ by activated CD4+ T cells [The flavone naringenin, found in citrus fruits, is characterized by its antioxidant properties and immunomodulatory effects ,116. Nar T cells .2+, and Lyn/NF-κB signaling [Kaempferol, abundant in spinach and kale , reducedignaling .4 was reduced in response to nobiletin, but not in response to tangeretin. Both nobiletin and tangeretin suppressed ERK1/2 phosphorylation upon IgE-mediated stimulation. The potentially affected pathway involved ERK in IgE-dependent stimulation and NF-κB in LPS-mediated activation [Citrus peel polymethoxyflavonoids, such as nobiletin and tangeretin , are a gtivation . In PMA tivation . Nobilettivation .2, TNF, and IL-6. Interestingly, low concentrations of resveratrol (<10 µM) enhanced TNF production in hsMC after FcεRI cross-linking [4 and PGD2 by more than 80%. The BMMC degranulation of β-hex was attenuated by about 60%. Resveratrol decreased the phosphorylation of protein tyrosine phosphatase 1B (PTB1B) and Syk, thereby inhibiting FcεRI-dependent MC activation by regulating the Syk pathway [Resveratrol belongs to the group of polyphenols, precisely the stilbenes , and is -linking . In cont pathway .In either IL-33- or IgE/antigen-stimulated BMMC, treatment with 25 µM resveratrol reduced the release of TNF-α, IL-13, and IL-6 by more than 40%. The underlying mechanism may be the regulation of the MAPK-activated protein kinase (MK)-2/3-PI3K/Akt axis, as IL-33-induced IL-6 and IL-13 production in MC is mediated by MK2/3 mediated activation of the PI3K/Akt pathway . RBL-2H3 cells stimulated with either IL-33  or IgE/aA higher dose of resveratrol (200 µM) showed a dose-dependent attenuation of C48/80-induced β-hex and histamine release by about 80% in the LAD2 MC line. Cytokine expression of MCP-1, TNF-α, and IL-1β was inhibited by resveratrol by about 40%, 60%, and 80%, respectively. Resveratrol administration increased the expression of nuclear erythroid 2-related factor 2 (Nrf2) and the generation of its target genes transcription factor HO-1 and NADPH dehydrogenase quinone 1 (NQO1). Thus, the Nrf2/HO-1 pathway may serve as a target for the therapy of MC-mediated allergic disorders . Using HIn an OVA-induced food allergy mouse model, resveratrol reduced the serum histamine and IgE levels, as well as MCP-1, by about 50% . We foun4 and PGD2 expression of about 80% [Curcumin and bisdemethoxycurcumin (BDMC) are two natural compounds found in curcuma, also known as turmeric. Both compounds have been shown to have anti-inflammatory properties ,129,130.bout 80% . Curcumibout 80% ,72,73,93bout 80% . These f2, and LTC4 (≈50%) levels in a mouse model of passive systematic anaphylaxis [The oral administration of curcumin reduced serum histamine, PGDphylaxis . Curcumiphylaxis ,92,93. Cphylaxis . ConversCinnamon extract (CE) and its compounds can be obtained from cinnamon bark and can have anti-inflammatory effects on MC ,75. CE t4 and CXCL8. IgE-mediated expression of CXCL8, CCL2, CCL3, and CCL4 were downregulated in hiMC. Similar inhibitory effects of CE and CA were observed, except for the expression of the MC proteases tryptase and chymase, which remained unaffected after CA treatment. ERK phosphorylation was reduced in both RBL-2H3 and hiMC, and PLCγ1 phosphorylation was additionally downregulated in RBL-2H3, both of which are important signaling proteins of MC activation [In addition, we found that cinnamaldehyde (CA) is the main mediator of CE in MC inhibition . RBL-2H3tivation . 4-chlortivation . 4-chlortivation . These Mtivation . Therefotivation .In addition to in vitro and in vivo studies, the effects of dietary components on allergic diseases have also been reported in randomized controlled trials (RCT) . Table 33) levels were increased by about 50%. Overall, adjuvant vitamin D therapy increased the effective rate of DCD from 84% to 97% [3 levels were elevated by 60% after vitamin D supplementation. In addition, adjuvant vitamin D supplementation reduced symptom severity in patients with AR. Interestingly, all significant effects were observed after eight weeks of treatment compared to four weeks of treatment, indicating the time span for vitamin D to induce its immunologic effects [Mild seasonal pollen allergic rhinitis (AR) treated with intranasal vitamin D adjuvant therapy showed a reduction in AR symptoms (≈40%), serum IL-4, and peripheral blood eosinophils compared to desloratadine citrate disodium (DCD) treatment alone. Serum calcidiol (25(OH)D% to 97% . Further effects . VitaminOral administration of the flavonoid quercetin to adult patients with allergic symptoms of AR resulted in a reduction in total symptom score (≈27%), sleeping disturbance (≈40%), and pruritus other than ocular/nasal. Further, the number of eosinophils in nasal discharge was reduced .O-methylated EGCG reduced symptoms of Japanese cedar pollinosis. Oral consumption of 700 mL green tea reduced symptoms of runny nose, itchy eyes, tearing, total nasal and ocular symptom score, and nasal and ocular symptom-medication score by less than 20% during the pollen season. Moreover, the peripheral eosinophil counts were reduced after pollen exposure [O-methylated EGCG may not be strong enough to be an alternative to pharmacological treatment. However, the daily use of Benifuuki may reduce the symptoms of AR [The daily consumption of Benifuuki green tea containing exposure . Althougms of AR .Adult AR patients treated with resveratrol showed a reduction in nasal symptoms compared to the placebo group. As a related effect, resveratrol reduced IgE, IL-4, TNF-α, and eosinophil levels in the blood of participants. Additionally, resveratrol treatment improved the quality of life of adults with AR . Further2 or LTC4 in PMN [The administration of curcumin in capsules to adults with perennial AR reduced nasal symptoms (≈60%) such as sneezing, itching, rhinorrhea, and nasal obstruction compared to the placebo group. In addition, curcumin decreased the cytokines levels of IL-4 and TNF-α in activated peripheral blood mononuclear cells (MNC) and IL-8 in activated polymorphonuclear neutrophils (PMN). Curcumin increased the production of IL-10 and soluble intercellular adhesion molecule-1 (sICAM-1), and improved the baseline nasal airflow in curcumin-treated patients with AR. However, curcumin had no effect on IFN-γ in MNC and PGE4 in PMN . Adult patients with acute symptoms of AR were treated with cinnamon bark extract. After seven days, patients showed a decrease in nasal, eye, and non-nose/eye symptoms. In addition, the total white blood cell count and neutrophil count decreased after treatment with cinnamon bark extract . In concAs the prevalence of allergic diseases continues to increase, appropriate therapeutic and preventive strategies against them need to be developed . In thisThe inhibitory effects of these nutrients on MC activation in vitro and in vivo are not always in the same range. Apart from their different effects on MC, different concentrations/triggers and MC/animal models were used depending on the experiment. This must be taken into account when comparing their effects on MC. In this review, we report that SCFA butyrate, vitamin D , polyphenols such as kaempferol, quercetin, luteolin, and resveratrol, as well as spices such as curcumin and cinnamon, exhibited the most potent inhibitory effects on MC degranulation, arachidonic acid metabolites, and cytokine release. These effects were observed in both in vitro and in vivo studies, highlighting the potential of these substances to modulate MC activity, and associated inflammatory responses. On average, the inhibitory effect of the dietary components on MC was lower in vivo than in vitro. In addition, these in vivo animal studies did not always clarify whether the dietary compounds inhibited MC directly or indirectly and what concentrations they reached in the MC. RCT were used to summarize the efficacy of the nutritional substances on MC-mediated allergic diseases.O-methylated EGCG, resveratrol, curcumin, and cinnamon on patients with AR could be included in this review. All of them showed an effect on the reduction of AR symptoms, inflammatory mediators, or immune cells. The oral administration of curcumin (≈60%) and nasal administration of cinnamon extract (≈50%) showed the strongest effects on the reduction of AR-specific symptoms [O-methylated EGCG, and resveratrol reduced allergy symptoms by less than 40%, with O-methylated EGCG having the weakest effect (<20%) [RCT on the effects of vitamin D, quercetin, symptoms ,143. Vitt (<20%) ,138,139.t (<20%) .In both included studies, vitamin D was used as a supplement to the conventional medication. Thus, the combination of vitamin D and the antihistamine drug (97%) resulted in a more effective improvement in relative symptoms in AR patients compared with the antihistamine drug alone (84%). It can be concluded that vitamin D can be used as a nutraceutical in the adjuvant therapy of AR ,137. Whi2 and LTC4 (≈50%) in in vitro and in vivo animal models [Curcumin showed inhibitory effects on the secretion of PGDl models , while nl models . To achil models , while, l models . Howeverl models .In this context, “bioavailability” refers to the fraction of the ingested nutraceutical that is accessible for absorption at the level of the gastrointestinal tract (GIT), metabolized, and distributed to organs and tissues in the GIT . Many suOne way to improve the poor bioavailability of nutraceuticals is to co-administer a potent bioenhancer, such as piperine, from black pepper . PiperinThe use of nanocarriers as delivery systems for nutraceuticals has been proven to be an effective method to increase the bioavailability of nutraceuticals . NanocarO-methylated EGCG, resveratrol, curcumin, and cinnamon on allergic rhinitis symptoms and inflammatory cell reduction. In particular, the oral administration of curcumin and nasal administration of cinnamon extract have been identified as potential substances to reduce allergic symptoms of allergic rhinitis. However, the use of the dietary components as nutraceuticals in allergic diseases is limited by their poor bioavailability. There are several strategies, such as nanocarriers as delivery systems, to favorably influence the pharmacokinetics of these nutrients. Further clinical studies are needed to investigate their oral bioavailability and delivery systems. Nevertheless, these dietary components could be considered as nutraceuticals in future studies as potential adjuvants or alternative medicines for the prevention or treatment of allergic diseases.The studies included in this overview suggest that dietary components such as fatty acids, amino acids, vitamins, carotenoids, flavonoids, and spices are able to attenuate proinflammatory, particularly IgE-dependent MC-mediated responses in vitro and in vivo. Overall, butyrate, vitamin D, the polyphenols kaempferol, quercetin, luteolin, and resveratrol, and the spices curcumin and cinnamon, were found to have the most potent anti-inflammatory effects on MC in vitro and in vivo. RCT reported beneficial effects of vitamin D, quercetin,


CBX2 and CEP55 in HCC may be caused by CNV alterations, promoter hypo-methylation, open chromatin accessibility, and greater active marks such as H3K4me3, H3K4me1, and H3K27ac. Functionally, CBX2, which was highly correlated with CD44, shaped a cancer stem cell-like phenotype by positively regulating cell-cycle progression, proliferation, invasion, metastasis, wound healing, and radiation resistance, revealed by combining bulk RNA-seq and scRNA-seq datasets. CBX2 knockdown validated its role in affecting the cell cycle. Importantly, we revealed CBX2 could activate gene by cooperating with co-regulators or not rather than a recognizer of the repressive mark H3K27me3. For instance, we uncovered CBX2 bound to promoter of CTNNB1 and CEP55 to augment their expressions. CBX2 showed a highly positive correlation with CEP55 at pan-cancer level. In addition, CBX2 and CEP55 may enhance extracellular matrix reprograming via cancer-associated fibroblast. Surprisingly, patients with high expression of CBX2 or CEP55 exhibited a higher response to immunotherapy, indicating that CBX2 and CEP55 may be promising therapeutic targets for HCC patients.Hepatocellular carcinoma (HCC) is a malignant tumor with a high prevalence and fatality rate. CBX2 has been demonstrated to impact the development and advancement of various cancers, albeit it has received limited attention in relation to HCC. In this study, CBX2 and CEP55 were screened out with the refined triple regulatory networks constructed by total RNA-seq datasets  and a robust prognostic model. Aberrantly higher expression levels of  Hepatocellular carcinoma (HCC) is the major cancer type of liver cancer. High mortality, high rates of recurrence, and high rates of metastasis are characteristics of hepatocellular carcinoma . TransarNEAT1 altered the miR-362-3p/MIOX axis to increase ferroptosis [Long non-coding RNA (lncRNA), a subtype of ncRNAs whose length greater than 200nt, could competitively bind to the miRNA, a kind of single-stranded ncRNA with length 18-25 nt, to regulate the expression of target, that is, the regular network , 5. In troptosis .DUXAP8, MCM3AP-AS1 and CDKN2B-AS1 are over-expressed in HCC, could be ceRNAs through miR-422a/PDK2, miR-194-5p/FOXA1, and let-7c-5p/NAP1L1 to promote HCC, respectively [DUXAP8/MCM3AP-AS1/CDKN2B-AS1 miR-424-5p/CBX2/CEP55 axis, first mentioned in HCC, was discovered in this study.ectively . The DUXCBX2 in HCC could induce a majority of genes to be silenced [CBX2, as a key member of PRC1, recognized the sites modified by the repressive mark H3K27me3 to prevent gene transcription, indicating the abnormal expression of silenced .CBX2 may enhance gene expression of CEP55 or CTNNB1 rather than the repressive function, especially the genes related to cell cycle, suggesting a crucial role of CBX2 for the cell cycle. Additionally, we have identified that CBX2 expression is cooperatively augmented by miR-425-5p, CNV alterations, DNA hypo-methylation, and higher active mark signals. CBX2 affected the drug sensitivity or sensitivity by regulating the cell cycle pathway. Importantly, despite having a worse prognosis and a more advanced stage or grade, patients with high levels of CBX2 or CEP55 expression may still benefit more from immunotherapy. Furthermore, CBX2 or CEP55 could potentially serve as therapeutic targets for pan-cancer.Here, we showed From the TCGA database, we downloaded the expression profiles (mRNA and miRNA) and clinical data of human hepatocellular carcinoma (LIHC). 367 HCC samples and 50 paired solid tumor-normal samples, as well as 371 HCC samples and 50 paired solid tumor-normal samples, respectively, made up the available mRNA and miRNA profiles. Downloaded from the GEO database was cohort GSE140845, whose miRNA count profile included 3 HCC samples connected to HCV and 3 paired normal samples. We performed quality control checks, read mapping operations, and finally generated the mRNA counts matrix using Fastp, HISAT2, and featureCounts, respectively, for the total RNA sequencing data in GSE140845 that were downloaded from SRP231432. The count matrices for mRNA, lncRNA, and miRNA were converted into TPM (transcripts per million) matrices and CPM (count per million mapped reads) matrices, respectively. MiRBaseVersions.db packages handled the conversion of immature miRNA names into mature miRNA names.CBX2, and CEP55.Then, using a count matrix and the DESeq2 R packages, we identified the differential lncRNAs, miRNAs, and mRNAs between these two cohorts with |logFC| 1 and FDR 0.01. Meanwhile, we also download the RNA-seq expression profiles from ICGC , GSE12455 (n=70), GSE138485 (n=64), GSE144269 (n=140), GSE148355 (n=69), GSE25599 (n=20), GSE55758 (n=16), GSE94660 (n=42), and the microarray expression profiles from GSE22058 (n=197), GSE25097 (n=511), to further validate the aberrant expression of CBX2 and CEP55.From the TCGA database, we also obtained the mutation status and CNV profiles for Initial regulatory networks. Given that lncRNA may compete with miRNA for binding sites, acting as a natural sponge to indirectly control mRNA expression. As a result, the initial regulatory network was created using the methods described below: (1) DElncRNA-DEmiRNA pairs were constructed using the lnc2base, ENCORI, and miRWalk2.0 databases; (2) DEmiRNA-DEmRNA pairs were predicted and confirmed using the miRWalk2.0, miRWalk3.0 databases, and miRNAtap packages; (3) We combined the DElncRNA-DEmiRNA pairs and the DEmiRNA-DEmRNA pairs to create the triple regulatory network under the following circumstances: (a) DElncRNA-DEmiRNA pairs had at least one supporting piece of evidence; (b) DEmiRNA-DEmRNA pairs had more than two supporting pieces of evidence; (c) up-regulated DElncRNA might down-regulate DEmiRNA to up-regulate DEmRNAs; and (d) down-regulated DElncRNA might up-regulate DEmiRNA to down-regulate DEmRNAs.Refined regulatory network. In the initial regulatory network, the Pearson correlation coefficient (PCC) between DElncRNA and DEmRNA was calculated. The PCC 0.3 from the initial regulatory network was used to define the refined regulatory network.In order to determine the hub regulatory network, we used the Cytoscape plug-in CytoHubba. Based on the sequences we obtained from LNCipedia, we then used lncLocator to predict the cellular location of DElncRNA. The initial regulatory network was visualized in Cytoscape, and the final regulatory network was done in R using the ggalluvial package.With the aid of the fgsea package, we conducted Gene Set Enrichment Analysis (GSEA) using signatures from MsigDB, such as Gene Ontology , KEGG (C2), and Hallmark (H). The statistical significance of enriched signatures was determined by the false discovery rate (FDR) adjusted P 0.05.CBX2/CEP55. The clusterProfiler package used the DEGs to investigate potential functional pathways or Gene Ontology .The DESeq2 package was used to find the differentially expressed genes related to CBX2/CEP55-related differentially expressed genes were discovered using the DESeq2 package. The DEGs were used by the clusterProfiler package to look into potential functional pathways or Gene Ontology .The Prognostic variables were chosen using the randomForest algorithm from the randomForestRSC package and the LASSO algorithm from the “glmnet” package. Candidates for prognostic DERNA served as the regression’s independent variable, and overall survival and overall status in TCGA-LIHC cohorts served as the response variables. Further PH testing was done on chosen prognostic RNAs. On the chosen, a multivariate Cox regression analysis was conducted and the ggforest package was used to visualize the results. RMS R package was used to confirm nomograms as well. The prognostic model’s reliability was verified both internally and externally. By randomly dividing the TCGA-LIHC into a training cohort and a validation cohort 50 times, internal validation was carried out. And for external validation, we used the ICGC-LIRI-JR expression profile. Calibration curves, the concordance index (C-index), and the time-dependent ROC curve were used to assess the prognostic model.The patients’ risk scores were determined using the normalized expression of confirmed prognostic RNAs and their corresponding regression coefficients in accordance with the prognostic model. Using the median risk score, the patients were then divided into high-risk and low-risk groups. To investigate the prognostic value of the risk score, univariate Cox regression analysis was used. In the Multiple Cox Regression Analysis, the risk score was combined with age, gender, and TNM stages.CBX2/CEP55 and their DNA methylation level. We also investigate the effect of the CBX2/CEP55 methylation level on overall survival.The TCGA database was used to download the Methylation array 450K profiles that correspond to the expression profiles. Other methylation profiles, including GSE44909 (n=24), GSE73003 (n=40), GSE82176 (n=19), GSE37988 (n=124), GSE55752 (n=16), and GSE113019 (n=60), were downloaded from GEO databases. The Spearman correlation coefficient was used to assess the relationship between The TCGA database was used to download the ATAC-seq profiles of the LIHC tumor samples, and the GSE173277 database was used to download the ATAC-seq data of the normal liver. Additionally, we downloaded the ATAC-seq data for the following cell lines from GEO: GSE172053 for Hep3B, GSE180143 for HepG2, GSE184796 for HepG2, GSE139190 for HepG2, GSE184797 for Huh7, and GSE192771 for Huh7. For the ATAC-seq analysis, we used Fastp for quality control steps, HISAT2 for read mapping onto GRCh38 reference steps, Samtools for bam file sorting, Macs2 for peak calling, and bdg2bw.sh for bedgraph file conversion into bw file. Finally, the WashU Epigenome Browser was used to visualize the tracks.Active marks H3K4me3, H3K3me1, and H3K27ac histone ChIP-seq datasets were downloaded from GSE112221 (liver tissues), GSE113879 (liver tissues), GSE76344 (HepG2), GSE184796 (HepG2), GSE92328 , GSE103730 (Huh7), GSE184797 , GSE113879 (Hep3B), GSE172053 (Hep3B), GSE208334 (PLC), GSE168178 (Hu1545). We used analysis techniques that were similar to those used for ATAC-seq datasets.Data for CBX2 primitive ChIP-seq was downloaded from the following GSEs: GSE29611 (K562), GSE59395 , and GSE34774 (293T). The analysis was conducted in the manner described above. IDR located the peaks that were shared. HOMER2 also identified the potential motifs and the clusterProfiler package was used to conduct enrichment analysis on the genes whose close peaks were located on the promoters.CBX2 knockout or knockdown, such as GSE54580 (AML), GSE193477 (AML), GSE112227 (Mouse bone), and GSE156413 (Mouse embryonic fibroblasts), were also gathered and analyzed.As previously mentioned, additional pre-existing data referred to as CBX2/CEP55 were examined.The immune, stromal, and TME scores were estimated using the ESTIMATE packages. The Tumor Immune Estimation Resource 2 portal (TIMER2) databases’ immune infiltration data, including CD4 T cells and CD8 T cells, were downloaded using the CIBERSORT, MCPCOUNTER, EPIC, and QUANTISEQ algorithms. Immune inhibitor genes, immune cell score, and the Spearman correlation between Based on the genome mutation maf files obtained from TCGA-LIHC, the tumor mutational burden (TMB) score was calculated. The Tumor Immune Dysregulation and Exclusion (TIDE) was used to predict the effectiveness of immunotherapy and indicated immune dysfunction and escape. A low TIDE score indicated good efficacy.The CancerSEA database  and ThorThe reverse phase protein array (RPPA) protein expression data were downloaded from the TCPA database. The GSCA database was used to download the pathway activity scores for the cancer-related pathways TSC/mTOR, RTK, RAS/MAPK, PI3K/AKT, hormone ER, hormone AR, EMT, DNA damage response, cell cycle, and apoptosis.Based on the transcriptomic profiles and drug sensitivity data in PRISM and CTRP2, we first used the oncoPredict package to train a drug sensitivity prediction model with tenfold cross-validation. The AUC value for each drug was then predicted using the fitted model and comparable transcriptomic data.A one-class logistic regression machine learning algorithm (OCLR), which built the model using training the human stem cell data from Progenitor Cell Biology Consortium (PCBC), calculated the mRNAsi scores of the TGCA-LIHC samples.Human Protein Atlas , GSE140228, GSE146115, GSE98638, GSE166635, GSE140228 and GSE125449 provided the scRNA datasets for HCC. The Seurat version 4 R package was used to process the scRNA datasets.https://www.proteinatlas.org/), a database of the human proteome. CBX2 and CEP55 immunohistochemical and immunofluorescent staining images in https://www.<soft-enter/>proteinatlas.org/). Detailed website information is provided in the corresponding Supplementary Figure Legend.Information on the distribution of proteins in human tissue and cells was available in the Human Protein Atlas  and GSE140845 , performed differential gene analysis, and found 6980 DEGs  in TCGA-LIHC and 843 DEGs . Finally, we established the shared genes responsible for the upregulation of 28 DElncRNA, 150 DEmRNA, and 6 DEmiRNA in HCC compared to 24 DElncRNA, 130 DEmRNA, and 2 DEmiRNA in adjacent tissue among these DEGs in two cohorts . HeatmapIn order to create the refined regulatory network, we first tried to create a basic regulatory network using one of two methods: (1) up-regulated lncRNA up-regulated mRNA by down-regulating miRNA; and (2) down-regulated lncRNA down-regulated mRNA by up-regulating miRNA. By at least one database of lnc2base, ENCORI, and miRWalk2.0, the predicted lncRNA-miRNA pairs in the initial network were verified. While at least two databases - ENCORI, miRWalk2.0, miRWalk3.0, and miRNAtap -were used to confirm the miRNA-mRNA pairs. Finally, using 37 DElncRNAs , 61 DEmRNAs , and 8 DEmiRNAs , the initial regulatory network was completed . We creaCBX2/CEP55/MCM2) and the second strategy (ESR1)  . Through1 in HCC . Fortuna miRanda   were significantly higher in the CBX2high tumor than the CBX2low tumor and the CEP55high tumor than the CEP55low tumor  between all genes and CBX2 or CEP55 in LIHC and LIRI-JP cohorts. We also identified the cell cycle pathway, DNA replication pathway, ribosome pathway, cell cycle-related hallmark , EMT, apoptosis, and WNT beta-catenin signaling using GSEA with the KEGG and hallmark gene set based on the PCC , GSE112227 (mouse long bone), and GSE156413 (mouse embryonic fibroblast). We discovered the cell cycle pathway and cell cycle-related pathways  were enriched in the CBX2 WT group  between the two genes and the cancer-related pathway score revealed that  pathway . Throughmote AML . EMT maymote AML .CBX2 and CEP55 may regulate the cell cycle process to support HCC.The overall result showed that CBX2 knockout reduced CEP55 expression , whose marker gene MKI67  strongly connected with CBX2 and CEP55, consistently with ECM enrichment , and tumor mutation burden (TMB) were considered new indicators of immunotherapy efficacy . Since Totherapy . In ordeotherapy . The Tumotherapy –13D. Addresponse  , Binimetinib (a MEK inhibitor), Voreloxin (a topoisomerase II inhibitor), Floxrudidine (oncology antimetabolites), Lenvatinib, Regorafenib, Paclitaxel, and Indisulam but not Sorafenib, demonstrating CBX2 affects the effectiveness of medications , in addition to miRNA miR-424-5p  and CBX2 . We used ChIP-seq in HepG2 to investigate how CBX2 improved the cell cycle. Surprisingly, we discovered that CBX2 could bind to the promoter of CEP55 and CTNNB1, increasing the expression of both genes (The cell cycle was kept in working order by th genes , 5D, 5E.th genes . This deCBX2high tumors, destroying the p53-p21-DREAM axis or the p53-p21-RB-E2F axis, as evidenced by the higher TP53 mutation rate [CBX2, not DREAM targets [CBX2 knockout prevented CCND2 expression, and CBX2 showed a negative correlation with CDKN1A, which encodes p21 (Furthermore, the p53 was dysregulated in ion rate , 37. Surodes p21 . This suCBX2 and CEP55 had an impact on the function states of cancer, especially the cell cycle. Additionally, CBX2 and CEP55 positively regulated the following processes: proliferation, metastasis, EMT, invasion, differentiation, hypoxia, wound healing, and apoptosis (CBX2 and CEP55 overexpression may lead to cancer stem cell-like phenotype and is supported by higher expression of CBX2 and CEP55 in cancer stem cells [CBX2high tumors (CBX2high tumors could be treated with Voreloxin, a Topoisomerase II inhibitor that targets TOP2A, whose expression was strongly correlated with CBX2 (As anticipated, poptosis , 14B. Them cells  (Supplemh tumors , 12E, inith CBX2 .CBX2high or CEP55high tumors displayed a highly complex tumor environment with both activated and suppressor cells (CBX2 was primarily found in hepatocytes or cancerous cells, where it drove the remodeling of the extracellular matrix and was significantly correlated with cancer-associated fibroblast. CBX2 or CEP55’s effects on TME need to be specifically investigated.Previous research suggested that immune infiltration may affect the patient’s prognosis . We founor cells , 14. AddCBX2 and CEP55 in HCC. Additionally, CBX2 may facilitate the cell cycle by directly working with co-regulators to control CEP55 and CTNNB1 or by indirectly competing with the DREAM complex. The phenotype that resembles cancer stem cells may be enhanced by the overexpression of CBX2 and CEP55. CBX2 and CEP55 may serve as potential drug targets and important genes for the effectiveness of immunotherapy. The triple regulatory networks predicted by the databases, the true impact of CBX2 on CEP55 or CTNNB1, and the repressive or active mechanism of CBX2 are a few limitations to be aware of. Therefore, additional fundamental research is needed to investigate the direct functional mechanism.In conclusion, using the refined regulatory network and dependable prognostic model, we verified the abnormal Supplementary FiguresSupplementary Table 1Supplementary Table 2Supplementary Tables 3 and 4


A Cox proportional-hazards regression analysis was conducted. Serum levels of IL-6, IL-1β, and IFN-γ were associated with lung cancer risk. IL-6 was associated with lung cancer risk, regardless of the histological type. IL-1β and IFN-γ had associations with adenocarcinoma and squamous-cell carcinoma, respectively. This study, targeting an Asian population, adds causality to association between proinflammatory cytokines and lung cancer risk by replicating previous findings. Furthermore, it suggests that specific cytokines, depending on the histological type of lung cancer, may serve as potential biomarkers.Recent studies have shed light on alterations in the proinflammatory tumor microenvironment as a significant carcinogenic mechanism. We carried out a case-cohort study to investigate associations between proinflammatory cytokines and lung cancer risk, considering histological types in the Korean general population . Serum levels of proinflammatory cytokines  were measured using Quantikine® ELISA. A Cox proportional-hazards regression analysis was conducted. In this study, serum levels of IL-6, IL-1β, and IFN-γ were associated with lung cancer risk. IL-6 was associated with lung cancer, regardless of the histological type. IL-1β had an association only with adenocarcinoma, while IFN-γ had an association only with squamous-cell carcinoma. This study shows associations between serum levels of IL-6, IL-1β, and IFN-γ and lung cancer risk, underscoring the potential of these cytokines to act as risk biomarkers. The utilization of these biomarkers for risk prediction may hold the promise of facilitating the identification of the high-risk population.Recent studies have shed light on alterations to the proinflammatory tumor microenvironment as a significant carcinogenic mechanism. Despite previous studies on associations between proinflammatory cytokines and lung cancer risk, few studies have been conducted in Asian populations. This study aimed to investigate associations between proinflammatory cytokines and lung cancer risk, considering histological types, in the Korean general population. We carried out a case-cohort study on the Korean National Cancer Center Community (KNCCC) cohort . Pre-diagnostic serum levels of proinflammatory cytokines  were measured using Quantikine Worldwide, lung cancer is the most common cause of cancer-related deaths, constituting 21.5% of all deaths in men and 13.7% of them in women, and it accounts for 14.3% of new cancer cases among men and 8.4% among women : 8.54 [3.30–22.11]), 3rd quartile . In addition, the lung cancer risk was associated with higher levels of IL-1β and IFN-γ than the LOD value  .After stratification analysis with the cigarette smoking status, the effect of serum proinflammatory cytokines on the lung cancer risk was more prominent in never-smokers. IL-6, IL-1β, and IFN-γ had consistent associations with the lung cancer risk , whereas, in ever-smokers, only IL-6 had an association with the lung cancer risk (HR [95% CI]:1.38 [1.16–1.64]) .The hazard ratios (HRs) are presented for each unit increase in natural log-transformed proinflammatory cytokines, derived from Cox proportional-hazards models. Model 1 was unadjusted; Model 2 was adjusted for sex; Model 3 was further adjusted for year of enrollment, region, education achievement, cigarette smoking status , number of pack-years (continuous variable), alcohol consumption status , and BMI (continuous variable). The following abbreviations are used: IL-6, interleukin 6; TNF-α, tumor necrosis factor-α; IL-1β, interleukin; IFN-γ, interferon gamma; and IL-10, interleukin 10.Our study demonstrates the association between serum levels of IL-6, IL-1β, and IFN-γ and the lung cancer risk. These associations persist even after adjustment for various potential confounding variables and are also strengthened by stratification for cigarette smoking status. Furthermore, in this study, the utilization of serum samples collected prior to the diagnosis of lung cancer negates the possibility of reverse causation, which implies that the presence of undiagnosed lung cancer might have influenced the serum levels of proinflammatory cytokines. It is important to note that we excluded cancer occurrences within 1 year of follow-up, and since the serum samples for measurement were collected a median of 6.79 years before the diagnosis of lung cancer, the robustness of the association between serum levels of proinflammatory cytokines and the lung cancer risk is supported.These findings replicate the observations of previous studies in Western populations and extend them to a general population in Korea. A study in the United States reported that both the National Cancer Institute—Maryland (NCI-MD) study and the Prostate, Lung, Colorectal, and Ovarian (PLCO) study reported increased odds ratios (ORs) for IL-6  and IL-8  . In a st7 and 1.4The findings of this study, as reported in previous studies ,13,14,15The role of inflammation and immune mechanisms in tumorigenesis is intricate. The disruption of the balance between Th-1 and Th-2 cytokines leads to increased inflammation, which results in elevated oxygen and/or nitrogen free radicals, which is linked to cancer development . Our stuDespite the meaningful findings of this study, there are some limitations. First, we only obtained serum samples at the time of enrollment in the cohort, and serum levels of proinflammatory cytokines were assessed only once. Consequently, we could not account for changes in proinflammatory cytokines induced by lifestyle modifications, such as smoking habits , alcohol consumption, dietary patterns, physical activity, and weight changes etc. Nevertheless, the associations among IL-6, IL-1β, and IFN-γ and the lung cancer risk in this study may not have been distorted, as these were not taken into account in both lung cancer cases and the subcohort. Second, our study encompasses several potential confounders, as occurs in most observational studies. To address this limitation, we conducted adjustments for age, sex, region, education achievement, cigarette smoking status, number of pack-years, and BMI. However, our findings might still be influenced by biases associated with other unmeasured confounders owing to the use of self-reported data and the potential existence of an unobserved confounder. Furthermore, in spite of our efforts to minimize the selection bias, the possibility for bias may persist. Third, the KNCCC cohort might not fully represent the Korean general population. However, it is important to emphasize that prospective cohort studies play a critical role in determining causal relationships between risk factors and diseases and may not necessarily require complete representativeness . FinallyIn conclusion, the findings of this study provide clinically valuable insights beyond the conventional questionnaires on tobacco history . In line


Neurological disorders affect the nervous system. Biochemical, structural, or electrical abnormalities in the spinal cord, brain, or other nerves lead to different symptoms, including muscle weakness, paralysis, poor coordination, seizures, loss of sensation, and pain. There are many recognized neurological diseases, like epilepsy, Alzheimer's disease (AD), Parkinson's disease (PD), multiple sclerosis (MS), stroke, autosomal recessive cerebellar ataxia 2 (ARCA2), Leber's hereditary optic neuropathy (LHON), and spinocerebellar ataxia autosomal recessive 9 (SCAR9). Different agents, such as coenzyme Q10 (CoQ10), exert neuroprotective effects against neuronal damage. Online databases, such as Scopus, Google Scholar, Web of Science, and PubMed/MEDLINE were systematically searched until December 2020 using keywords, including review, neurological disorders, and CoQ10. CoQ10 is endogenously produced in the body and also can be found in supplements or foods. CoQ10 has antioxidant and anti-inflammatory effects and plays a role in energy production and mitochondria stabilization, which are mechanisms, by which CoQ10 exerts its neuroprotective effects. Thus, in this review, we discussed the association between CoQ10 and neurological diseases, including AD, depression, MS, epilepsy, PD, LHON, ARCA2, SCAR9, and stroke. In addition, new therapeutic targets were introduced for the next drug discoveries. Neurological diseases endanger human health and lifestyle. Neurological diseases affect a large number of people all over the world H oxidoreductases in the plasma membrane, like NAD(P)H: quinone oxidoreductase 1, NADH-cytochrome b5 reductase, or NADPH-CoQ reductase  and ethanol (EtOH), before and during exposure to the parkinsonian neurotoxin, was rescued by both βNF and EtOH treatments. Neuroprotective effect of CYP inducers was due to a decrease in ROS production, restoration of mitochondrial fusion kinetics, and mitochondrial membrane potential  mutations result in secondary CoQ10 deficiency. CoQ5 and CoQ9 proteins are found in many mitochondrial protein complexes in human 143B cells and CoQ9 and CoQ5 knockdown inhibits CoQ10 levels  activity, while PDSS2 levels showed a positive correlation with malignancy. Also, lower mitochondrial DNA-encoded cytochrome c oxidase subunit 2 levels showed no association with a higher malignancy degree and lower CoQ protein levels. Mitochondrial abnormalities are linked to defected CoQ10 maintenance in the progression of human astrocytoma , protein oxidation, and mutagenesis . CoQ molecules are dynamically divided in a pool attached to and engulfed by the super-complexes I + III, and another pool related to complex II or other mitochondrial enzymes using CoQ as a cofactor. Such a CoQ-free pool can be applied by enzymes linking the MRC to other pathways, like the fatty acid β-oxidation and amino acid catabolism, pyrimidine de novo biosynthesis, proline, arginine, and glyoxylate metabolism, glycine metabolism, and sulfide oxidation metabolism that some of them are attached to metabolic pathways in other compartments  cerebellar ataxia, (2) severe infantile multisystemic disease, (3) encephalomyopathy, (4) isolated myopathy, and (5) nephrotic syndrome. In some cases, pathogenic mutations are observed in genes associated with the CoQ10 biosynthesis (primary CoQ10 deficiencies) or those not directly associated with CoQ10 biosynthesis (secondary CoQ10 deficiencies). The pathogenesis of primary CoQ10 deficiencies has been linked to respiratory chain defects, ROS generation, and apoptosis variably  and lipid peroxide concentrations in cultured cells, and antioxidants, like vitamin E with a protective role in neurons against Aβ-related cytotoxicity  of the hippocampal synaptic transmission  reduced LPO and elevated GSH levels and total antioxidant capacity (TAC) values, and SOD and glutathione peroxidase (GPx) activities in both brain areas of mice. The suppressed neuro-inflammatory response in the prefrontal cortex (PFC) and HIP was observed, evidenced by reduced NF-kB, p38, and JNK concentrations in the CoQ10 groups GSK-3β, CREB, pCREB, and BDNF protein expression in rats receiving chronic unpredictable mild stress (CUMS) for 6 weeks  on behavioral dysfunction and CoQ10 levels in the rat brain. A significant difference was found between the depression induced by streptozotocin (STZ) and control groups tested by the splash test and FST 24 h after STZ treatment. In addition, according to the validated and accurate high-performance liquid chromatography (HPLC), reduced CoQ10 levels were found in the brain of the STZ group  or/and fluoxetine (10 mg/kg/day) was assessed on mRNA expression, 5-HTEpilepsy is the commonest neurological disease worldwide, in which spontaneous unusual electrical discharges of neurons are observed throughout the brain Patel, . Temporain vitro  is a regulator of seizure activity because of its various anticonvulsant  increased the total number of spike-wave discharges (SWDs) but did not change the mean duration of SWDs. CoQ10 (100 and 200 mg/kg) increased both the total number and the mean duration of SWDs abolished by coadministration of 7-nitroimidazole . Coadministration of l-arginine   enhanced the CoQ10 effect on the total number of SWDs but not on its mean duration. Therefore, the effect of CoQ10 on seizure was attenuated by the NOS inhibitor. Furthermore, based on the electrophysiological evidence, CoQ10 administration increased the absence seizures through the stimulation of the neuronal NOS  enhanced time latencies to the onset of myoclonic jerks and clonic seizures induced by intraperitoneal pentylenetetrazole (PTZ) and at the doses of 25 mg/kg or more augmented the seizure threshold provoked by intravenous injection of PTZ. Subchronic doses of CoQ10 (50 mg/kg or more) reduced the tonic seizures induced by PTZ or electroshock. This antiseizure effect of subchronic CoQ10 was attenuated by the NOS inhibitor , neural and axonal injury, and oxidative stress are the causes of MS , progressive-relapsing MS, primary progressive MS, and secondary progressive MS are different MS types  by CoQ10 compared to controls. Also, the TNF-α level showed a significant decrease following CoQ10 administration (10 mg/kg/three weeks) vs. IL-10. The TH1/TH2 ratio in CoQ10-treated animals showed a significant decrease than in non-treated animals (P < 0.01)  and a reduction in MDA concentrations (P = 0.003) than controls. Although CoQ10 supplementation significantly affected plasma TAC values (p = 0.010), no significant difference was found between both groups. CoQ10 supplementation had no effect on GPx activity  neurons in the SNpc. Over 95% of PD patients are sporadic and PD is developed in cases older than 60 years  administration is the initial treatment for PD Nutt, . DiminutCurrently, PD cannot be cured; however, neuroprotectants reduce the rate of neurodegeneration and improve the quality of life  was assessed in the DJ-1/1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model of PD through histochemical and behavioral approaches. Application of Ubisol-Q10 could remarkably offset the neurotoxicity and ameliorate motor impairment by MPTP  showed significant effectiveness in chlorpromazine -induced Parkinsonism-like alterations in mice  , Kearns-Sayre syndrome (KSS), and other mitochondrial encephalomyopathies  gene as an ortholog of yeast coq8 (Lagier-Tourenne et al., The purpose of this article was to review the studies using experimental and clinical treatments . ExperimCoQ10 as an antioxidant and neuroprotective agent can play a role in the treatment of neurological disorders. Although neurological diseases cannot be treated effectively, CoQ10 deficiency is involved in the pathogenesis of epilepsy, stroke, MS, depression, PD, AD, LHON, ARCA2, and SCAR9. More clinical and experimental studies are needed using electrophysiological and behavioral evaluation, genetic targeting, and molecular imaging.SB, RH, SS, MK-A, AM, MR, and AK literature review and drafting the manuscript. SB and AK critically revision of the manuscript. All authors read and approved of the final manuscript.


Targeting kinase activity is considered to be an attractive therapeutic strategy to overcome acute myeloid leukemia (AML) since aberrant activation of the kinase pathway plays a pivotal role in leukemogenesis through abnormal cell proliferation and differentiation block. Although clinical trials for kinase modulators as single agents remain scarce, combination therapies are an area of therapeutic interest. In this review, the author summarizes attractive kinase pathways for therapeutic targets and the combination strategies for these pathways. Specifically, the review focuses on combination therapies targeting the FLT3 pathways, as well as PI3K/AKT/mTOR, CDK and CHK1 pathways. From a literature review, combination therapies with the kinase inhibitors appear more promising than monotherapies with individual agents. Therefore, the development of efficient combination therapies with kinase inhibitors may result in effective therapeutic strategies for AML. Acute myeloid leukemia (AML) is a clonal disease that is derived from abnormally and, occasionally, poorly differentiated cells of the hematopoietic system . ImproveAccumulated evidences suggests that abnormal activation of signal transduction pathways plays a pivotal role in leukemogenesis, through the blocking of differentiation and abnormal cell proliferation . Kinase To follow this issue in the present review, the author summarizes attractive kinase pathways for therapeutic targets and the combination strategies for these pathways. During a search of the literature on PubMed with the keywords “kinase”, “inhibitor”, “combination”, “AML”, “therapy”, and “clinical”, more than 500 papers were found to have been published to date. Based on the search, the FLT3, PI3K/AKT/mTOR, MAPK, AXL, CDK, and CHK1 pathways appear to be common pathways targeted by kinase inhibitor combination therapies. In the present review, the author summarizes these attractive kinase pathways for therapeutic targets and the combination strategies for these pathways.Mutations of FLT3 comprise one of the most frequently identified types of genetic alterations in AML . As the p = 0.002) and reduced relapse rates (p = 0.02); it was also remarkably advantageous for FLT3-ITD high-allelic ratio patients (2-year OS of 82%) [Several resistance mechanisms for FLT3 inhibitor therapy have been shown, such as additional mutations in the kinase domain in internal tandem duplication (ITD) patients , or anot of 82%) .FLT3 inhibitors are tyrosine kinase inhibitors and are classified into first- and second-generation inhibitors based on their kinase specificity and potency . First-gAs there are many excellent reviews on FLT3 inhibitors and their combinations ,27,28,29This combination was shown to be well tolerated in older patients with untreated FLT3-ITD AML , or undeThe phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin (PI3K/AKT/mTOR) signaling pathway is one of the key aberrant intracellular axes in AML . ChemothBertacchini et al.  investigAnother kinase pathway related to AKT/mTOR signaling involves PIM kinases and p38a (MAPK14) . SignaliMeja et al.  found thRecently, metformin, a classic hypoglycemic drug for diabetes, was reported to synergistically sensitize AML cells to Ara-C through mTORC1/P70S6K pathway inhibition . However, concerns about combinations with PI3K/AKT inhibitors have been raised. Liang et al.  revealedClinical trials on mTOR inhibitors have been conducted. In the early 2010s, the results of the phase Ib GOELAMS study of the mTOR inhibitor RAD001  and the One of the most aberrantly activated oncogenic pathways in AML is the RAS-RAF-MEK-ERK (MAPK) pathway . HoweverMAPK-interacting kinases 1 and 2 (MNK1/2) are downstream effectors of this pathway that control the activation of eukaryotic translation factor 4E (eIF4E)  Figure . eIF4E wAntiapoptotic Bcl-2 family members are critical for the survival of AML cells. The combination of venetoclax and tomivosertib showed synergistic anti-leukemic responses in AML cells . In a siAXL, named after the Greek word “anexelekto”, meaning uncontrolled, is a member of the TAM family of receptor tyrosine kinases. AXL potentially drives cell proliferation through effector molecules in the PI3K/AKT/mTOR, RAS/RAF/MEK/ERK, JAK/STAT, and NF-kB signaling pathways . InhibitC Cyclin-dependent kinases (CDKs) 1, 2, 4, and 6 are mainly involved in regulation of the cell cycle, while CDK7, 8, and 9 play roles in regulating transcription to further influence survival and cell proliferation by driving the target gene expressions . Among tCDK9 is a transcriptional regulator of myeloid cell leukemia-1 (MCL-1) that can influence apoptosis induction ,74. CDK9Recently, histone methyltransferase EZH2 loss and the subsequent reduction in trimethylation of histone H3K27 were reported to result in the de-repression of HOX genes as a novel pathway for acquired resistance to tyrosine kinase inhibitors and cytotoxic drugs in AML . SpecifiThe DNA damage checkpoint is regulated by two signaling pathways, ATM-CHK2-p53 and ATR-CHK1-cdc25A, of which the ATM-CHK2-p53 pathway is impaired in many cancers  Figure . ConsequA major cause of treatment failure is resistance to chemotherapeutic agents, and one strategy to overcome such chemoresistance is to target the antiapoptotic Bcl-2 protein. The Bcl-2-selective inhibitor ABT-199 showed encouraging preclinical results ,89. Mcl-


Background: Gastric cancer (GC) is one of the most common malignancies, with a low 5-year survival rate. However, if diagnosed at an early stage, it can be cured by endoscopic treatment and has a good prognosis. While gastrointestinal X-ray and upper endoscopy are used as national GC screening methods in some GC high-risk countries, such as Japan and Korea, their radiation exposure, invasiveness, and high cost suggest that they are not the optimal tools for early detection of GC in many countries. Therefore, a cost-effective, and highly accurate method for GC early detection is urgently needed in clinical settings. DNA methylation plays a key role in cancer progression and metastasis and has been demonstrated as a promising marker for cancer early detection.Aims and methods: This review provides a comprehensive overview of the current status of DNA methylation markers associated with GC, the assays developed for GC early detection, challenges in methylation marker discovery and application, and the future prospects of utilizing methylation markers for early detection of GC. Through our analysis, we found that the currently reported DNA methylation markers related to GC are mainly in the early discovery stage. Most of them have only been evaluated in tissue samples. The majority of non-invasive assays developed based on blood lack standardized sampling protocols, pre-analytical procedures, and multicenter validation, and they exhibit insufficient sensitivity for early-stage GC detection. Meanwhile, the reported GC DNA methylation markers are generally considered pan-cancer markers.Conclusion: Therefore, future endeavors should focus on identifying additional methylation markers specific to GC and establishing non-invasive diagnostic assays that rely on these markers. These assays should undergo multicenter, large-scale prospective validation in diverse populations. Helicobacter pylori and serum pepsinogen), have insufficient sensitivity and specificity (Gastric cancer (GC) is a major global health concern, it caused 1,089,103 new cases and 768,793 new deaths in 2020, ranking fifth for incidence and fourth for mortality among all cancer types globally . Howevercificity .SPET9 methylation test (Epi proColon) (DNA methylation is the most widely studied epigenetic modification which plays a significant role in cancer progression and metastasis . Many ofroColon) , and theroColon) , were suroColon) . MoreoveroColon) . PromoteroColon) , thus DNThis review provides a comprehensive overview of the DNA methylation changes in GC, and summarized the achievements, challenges and possible further direction for DNA methylation as a potential tool for GC early detection.In this review, we performed an extensive search and analysis of previously published DNA methylation markers to evaluate their effectiveness in the detection of GC across different sample types ; Table 3The methods used for analyzing DNA methylation markers included methylation-specific PCR (MSP), nested MSP, quantitative methylation-specific PCR (qMSP), Sanger sequencing, combined restriction digital PCR (CORD), methylation-sensitive melt curve analysis (MS-MCA), pyrosequencing and bisulfite sequencing ; Table 3P16, RNF180, RUNX3, SEPT9, E-cadherin, Reprimo, ELMO1, TFPI2, RASSF1A, SFRP2, RPRM, ZIC1, and DAPK) were reported at least 2 times. Among these markers, 37 out of them were analyzed in blood samples (plasma or serum), and the repeatedly evaluated markers were also assessed in blood samples at least once. In addition to blood samples, this review also included markers evaluated in tissue and gastric washes/juice. For instance, Sox17  were solely evaluated in tissue samples (fresh frozen tissue or FFPE). The remaining six markers  were evaluated simultaneously in both tissue and gastric wash samples  and specificities (94.6% and 97.0%) in 0.2 mL plasma, while another study demonstrated that RASSF1A had only 33.3% sensitivity with a specificity of 100.0% in 2 mL plasma  and intestinal metaplasia (IM)  . Other m)  of 0.84 for GC detection, indicating its potential as a non-invasive blood-based method for early detection of GC  and esophagogastric junction cancer (EJC) together, and achieved 67.9% and 71.0% sensitivities for GC and all cancer types, respectively, with a specificity of 90.0%. RASSF2 and SFRP2 were combined using for detection of CRC and GC in 0.1 g stool samples , and pyrosequencing  Kurdyuk. The entSEPT9 is approved by the FDA and NMPA as a plasma marker for CRC detection (SFRP2 (KCNQ5 (C9orf50 (CLIP4 (TFPI2 (ZNF582 (ZNF569 (RASSF1A has been approved by NMPA as a lung cancer early detection marker (In the field of GC early detection, the current methylation markers being used are predominantly pan-cancer markers rather than GC-specific markers. For example, etection , while Sn (SFRP2 , KCNQ5 (2 (KCNQ5 , C9orf50(C9orf50 , CLIP4 (0 (CLIP4  and TFPI4 (TFPI2  were fou (ZNF582  and ZNF5 (ZNF569  showed hn marker . MeanwhiCompared to DNA methylation in CRC early detection , the recIn conclusion, DNA methylation as a robust and sensitive marker has also been widely studied in GC samples, numerous potential DNA methylation markers have already been identified in GC, and some of them have been developed as commercial kits. But the lack of early-stage GC sensitivity and specificity should be improved in the future, and the standardized sampling, pre-analytical, cfDNA isolation and conversion procedures must be considered in development of the assay. With the rapid development of new technology and the discovery of more methylation markers, it is expected that DNA methylation will become a cost-effective and non-invasive tool for GC early detection in the near future.


The dysfunction of astrocytes in response to environmental factors contributes to many neurological diseases by impacting neuroinflammation responses, glutamate and ion homeostasis, and cholesterol and sphingolipid metabolism, which calls for comprehensive and high-resolution analysis. However, single-cell transcriptome analyses of astrocytes have been hampered by the sparseness of human brain specimens. Here, we demonstrate how large-scale integration of multi-omics data, including single-cell and spatial transcriptomic and proteomic data, overcomes these limitations. We created a single-cell transcriptomic dataset of human brains by integration, consensus annotation, and analyzing 302 publicly available single-cell RNA-sequencing (scRNA-seq) datasets, highlighting the power to resolve previously unidentifiable astrocyte subpopulations. The resulting dataset includes nearly one million cells that span a wide variety of diseases, including Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), multiple sclerosis (MS), epilepsy (Epi), and chronic traumatic encephalopathy (CTE). We profiled the astrocytes at three levels, subtype compositions, regulatory modules, and cell–cell communications, and comprehensively depicted the heterogeneity of pathological astrocytes. We constructed seven transcriptomic modules that are involved in the onset and progress of disease development, such as the M2 ECM and M4 stress modules. We validated that the M2 ECM module could furnish potential markers for AD early diagnosis at both the transcriptome and protein levels. In order to accomplish a high-resolution, local identification of astrocyte subtypes, we also carried out a spatial transcriptome analysis of mouse brains using the integrated dataset as a reference. We found that astrocyte subtypes are regionally heterogeneous. We identified dynamic cell–cell interactions in different disorders and found that astrocytes participate in key signaling pathways, such as NRG3-ERBB4, in epilepsy. Our work supports the utility of large-scale integration of single-cell transcriptomic data, which offers new insights into underlying multiple CNS disease mechanisms where astrocytes are involved. Astrocytes, one of the most abundant glia cell types in the central nervous system (CNS), have been implicated in the pathogenesis of multiple CNS disorders . AccumulAmong these investigations, high-throughput single-cell RNA sequencing (scRNA-seq) has been the leading method to characterize the cellular heterogeneity in neurological disorders, and its adoption has led to the generation of a plethora of datasets ,10,11,12Large-scale data integration can capture the comprehensive phenotypic features at single-cell resolution to empower the understanding of disease mechanisms ,15,16. OWe collected single-cell transcriptomic, spatial transcriptomic, and proteomic datasets originating from 17 well-generated studies published from June 2019 to March 2022. Some of these samples were donated by healthy individuals, and some were taken from autopsy tissues. In single-cell transcriptome data, except for Wheeler2020, which used scRNA-seq, all others used single-cell nuclear sequencing (snRNA-seq), which ensured the stability of the sequencing method. Similarly, except for Chancellor2021, which used the inDrops sequencing platform, the rest of the data were sequenced using the 10x Genomics platform. The age of the patients we collected ranged from 24 to 102 years old. As our study covers multiple brain regions, even the cortex alone includes more fine-grained subregions such as occipital, occipitotemporal, somatosensory, entorhinal, and dorsolateral cortexes. However, since some of the papers did not specify the exact brain region, we grouped them into four categories, cortex (CTX), hippocampus (HIP), white matter (WM), and basal ganglia (BG), for the analysis. The basal ganglia mainly include the three brain regions of caudate, putamen, and substantia nigra. We named the original brain region sources as ‘source.tissue’, and the new classification is annotated as ‘current.tissue’ in the paper. See We downloaded all raw scRNA-seq matrix data from different sources . We follp-value distribution for each principal component. Our initial results from K-nearest neighbor clustering, using FindNeighbors and FindClusters with the resolution = 0.2, and UMAP clustering, using RunUMAP with dim parameter set to 20, were 53 clusters. We found a significant batch effect in the merged datasets, so we performed a subsequent correction for the batch effect.After preprocessing and quality control, we merged all datasets using the Seurat::merge function. The UMI count matrix was log normalized, and the 3000 most variable features were identified. The linear dimensionality was reduced using the RunPCA function in Seurat. For graph-based clustering, we used the first 30 principal components determined by the ElbowPlot function, which displayed the GFAP and AQP4 for astrocyte; ABCB1 and FLT1 for endothelial cells; RBFOX3 and GRIN1 for neurons; CAMK2A and CBLN2 for excitatory neurons; GAD1 and GAD2 for inhibitory neurons; DOCK8 and CSF1R for microglias; MBP and PLP1 for oligodendrocytes; PDGFRA and VCAN for OPCs; and PDGFRB for pericytes.To remove batch effects and non-biological technical biases, the Harmony algorithm in Seurat was used. We have used default values for the other parameters of Harmony . This wap-value < 0.05 using the Bonferroni correction) was used to determine unique and/or highly enriched DEGs in one cluster compared to all other clusters. These cluster-specific features were used as reference genes of GO pathway analysis with the R package clusterProfiler  [We re-clustered astrocytes  and identified eight transcriptionally unique subpopulations. To identify the enriched genes in each astrocyte subtype, the FindAllMarkers function  . Data weASPG, S1PR3, CXCL10, VIM, GFAP, CD44, CP, OSMR, TGM1, FKBP5, TIMP1, HSBP1, and STEAP4); A1, A1-reactive astrocytes ; A2, A2-reactive astrocytes ; HM, homeostasis ; and PC, phagocytosis (MEGF10 and MERTK) [We assessed signature gene scores of astrocytes using the AddModuleScore function in Seurat. These genes originate from previously published studies including PR, pan-reactive astrocytes (d MERTK) ,26,27.We downloaded the recently published 10x Visium dataset from whole hemisphere saline or LPS-injected mice (N = 3 per condition) (data available at the Gene Expression Omnibus repository: series GSE165098) . In shorMef2c, Mef2d), hippocampus , white matter , and ventricle modules  [https://www.gsea-msigdb.org/gsea/msigdb/, accessed on 11 April 2023).We assessed module scores of classical region-specific genes using the AddModuleScore function in Seurat. These modules include cortex  ,30,31,32To highlight clusters identified in our single-cell RNA-seq data, we defined gene signatures for each astrocyte cluster using the top 50 marker genes (genes ranked by log2FC) from our earlier enriched genes in each astrocyte subtype . The specific steps are as follows. We first filtered DEGs of each astrocyte cluster . We used these genes to construct a normalized expression matrix (genes x cells) for each sample. If the number of astrocytes per sample was greater than 300, we set the number of decomposition rank to 8, and if it was between 100 and 300 then it was set to 6, which resulted in a total of 1556 submodules across the 294 samples. The 1556 submodules were compared by hierarchical clustering using the Pearson correlation coefficient over all gene scores as a distance metric. Based on the clustering dendrogram and cutree function in R, seven clusters of modules were identified manually. We used the top 20 genes of each module to define the module. For a single cell, if it expressed ≥70% of the genes within a given module, it was considered as a cell expressing the activated module, namely, a module cell. The samples with less than 100 astrocytes were excluded. For each of the 294 samples with at least 100 astrocytes, we used non-negative matrix factorization (NMF) to extract 7 co-expression submodules with the NNLM R package (version 0.4.4)  package (version 1.42.0) with standard settings . To assehttps://string-db.org, accessed on 11 April 2023) and used Cytoscape (version 3.9.1) to visualize the networks.We used the top 50 M2 and M4 modular genes to construct the interaction networks by STRING analysis  and used Cytoscape (version 3.9.1) to visualize the networks. PCA was performed based on the top 200 DEGs for each disorder using the scatterplot3d package (version 0.3-41).To avoid the spatial bias, the DEG analysis was firstly conducted between disease and controls from the same brain region. The number of unique and overlapping up- and down-regulated DEGs among different disorders are shown using UpsetR (version 1.4.0) . We usedp-values were indicated by a heatmap.We downloaded a recently published TMT-MS dataset, which has been normalized . We perfLigand-receptor analysis and visualization were performed using CellChat (version 1.1.3) . CellChaThe overall strategy includes collecting human scRNA-seq, spatial transcriptomic, and proteomic datasets to avoid the limitations by single methodology A–C. BrieADGRV1, AQP4, SLC1A2, and GFAP , suggesting their important putative protective roles in maintaining CNS homeostasis. In addition, cluster 0 was also involved in transition metal ion homeostasis , and expressed APOE and CLU involved in amyloid metabolism and clearance [SNHG14 and MIR100HG, were enriched in cluster 1. SNHG14 has previously been reported to provide neuroprotection by inhibiting inflammatory responses during AD progress [MEGF10, MERTK) were significantly enriched in cluster 1, which has previously been reported as a novel role for astrocytes in mediating synapse elimination to achieve precise neural connectivity [In total, 169,730 astrocytes were identified by showing high expression of markers such as and GFAP E. We souand GFAP F–H. The and GFAP G. Their and GFAP . We founlearance ,39,40. Iprogress . In addiectivity ,43. TherGFAP and CD44) were enriched in cluster 2 and 7, representing astrocyte activation [FOS, JUN/JUNB, UBC, and ID3) and the extracellular matrix  and associated with proteostasis . These reactive genes presenting in the same astrocyte cluster suggest their putative close interaction. Cluster 5 was enriched for inflammation/A1 astrocyte signatures and highly expressed immune response genes including DOCK8, APBB1IP, C3, SPP1, RUNX1, and PLXDC2. Previously published studies have reported complement component 3 (C3) in a specific subtype of reactive astrocytes that respond to inflammation and mediate microglia–astrocyte crosstalk in a range of neurodegenerative diseases such as AD, HD, MS, and amyotrophic lateral sclerosis (ALS) [MBP and PLP1) and higher OLIG2 (oligodendrocyte transcription factor 2) compared with other clusters  and was involved in synaptic maintenance. Together, we identified eight transcriptionally distinct astrocyte subtypes, suggesting that integrated data enabled us to detect novel subpopulations that had previously been missed.Pan-reactive astrocyte signatures  ,46. Clustrocytes . Therefo neurons ,48. LastRORB and TSHD7A were enriched, which have previously been reported to show cortex-specific expression in astrocytes and neurons [Recent findings showed astrocytes varied across the CNS regions ,11,49. W neurons  , hippocampus , white matter , midbrain , and ventricle modules  [TNC, CD44, and APOE), cluster 7 also highly expressed LUZP2 (log2FC = 2.48) compared to other clusters  ,30,31,32and Ttr) ,51. Meanand Ttr) . Therefoclusters . This isf the BG ,55.In addition, we also explored whether neuroinflammation induced by the endotoxin lipopolysaccharide (LPS) affects the expression of module signatures. We found that all the module signatures, except for cluster 4, were significantly upregulated in neuroinflammatory mice compared to normal controls E. TogethTo investigate how astrocytes contribute to disease onset, we preliminarily analyzed the relationship between astrocyte subtypes and disease . We founUnfortunately, we could not find statistical differences in AD compared with control groups, which is contrary to previous studies ,56. We hGPC5, RYR3, SLC4A4, and GABRB1). M1 module cells were involved in calcium ion transport, inorganic ion transmembrane transport, and regulation of cellular PH pathways , which comprise two AD-risk genes . M6 module cells also displayed small molecule catabolic process, lipid catabolic process, and amyloid beta clearance  and protein kinases (DCLK1/2) that were involved in radial migration and axon growth of cortical neurons and associated with neurodevelopmental and neuropsychiatric disorders [GLIS3 in the M2 module has been studied to influence the glioma cells’ invasion, migration, and proliferation and upregulate the NF-κB signaling pathway [HSPB1, HSP1A1, and HSPBB) and cellular stress . M4 stress cells showed activation of the response to oxidative stress, P53 pathway, apoptosis, and TNFα signaling (We used non-negative matrix factorization (NMF) for 294 human brain scRNA-seq datasets to uncover coherent sets of genes that were preferentially co-expressed by subsets of astrocytes. We identified seven expression modules (M1–M7) with different functions and cell statuses  and defipathways D. The M6learance D. These isorders ,59. Addi pathway . The M4 ignaling D. IL1RAPL1, CTNNA3, PTPRD, and DLG2) and ensheathment of neurons , in line with displaying regulation of axonogenesis and presynaptic organization pathways  and performed immune activation functions, including positive regulation of phagocytosis and the antigen receptor-mediated signaling pathway [CFAP43/54, DNAH7/9, HYDIN, and RFX3) and microtubule proteins . Together, we identified seven co-expression modules of astrocytes from multiple samples, which perform specific functions.In contrast, other modules were expressed in different astrocyte clusters in a special-cluster manner, such as M3 in cluster 3, M5 in cluster 5, and M7 in cluster 4 C. The M3pathways B,D. The  pathway ,62. The p < 0.05, We next explored how these modules are co-regulated in pairwise using Spearman correlation analysis E. We ideTo study whether astrocytes have contextual changes across different CNS disorders, we next analyzed differentially expressed genes (DEGs) and identified both common and disorder-specific transcriptomic changes across different CNS disorders including HD, PD, AD, Epi, CTE, and MS. To avoid spatial bias, the DEG analysis was conducted between disease and controls from the same brain region. In total, we identified 2294 upregulated DEGs and 3407 down-regulated DEGs across all disorders that were presumably driven by disorders . PrincipGFAP, FOS, and heat-shock genes. This network also suggested that many astrocytic genes are associated with multiple pathological processes including MAFG, CLU, APP, and ITM2B/C. Specifically, a recent study identified that MAFG-driven astrocytes promote CNS inflammation and repress antioxidant and anti-inflammatory transcriptional modules in MS and its preclinical model, experimental autoimmune encephalomyelitis (EAE) . Here, we further revealed that in addition to MS, HD, and PD, astrocytes also highly express MAFG. Given that comparatively less is known about the role of MAFG in other CNS diseases, we hypothesize that the same pathological mechanisms may exist in the other two diseases. We further explored transcriptomic differences across different CNS disorders and found that transcriptional changes associated with astrocyte reactivity are highly heterogeneous in a disorder-specific manner. The number of up-regulated DEGs varies greatly across disorders: MS (1315), epilepsy (1203), PD (1082), HD (639), AD (84), and CTE (76) B. Their EPHB1, PTN, NRP1, NRXN1, and GABRA2, which are involved in synapse organization, axonogenesis, and positive regulation of glial cell differentiation (EphB1) in a mouse ALS model and pleiotrophin (PTN) in MS could reduce pro-inflammatory signaling in astrocytes and promote neuronal survival following inflammatory challenge [For the down-regulated genes, PD, Epi, HD, and MS had more DEGs compared to AD and CTE disorders C. A totantiation . The lowhallenge ,72. We fTo explore whether there are some associations between the previously constructed modules and the diseases, we firstly calculated co-expressed module scores for the top 20 genes of each module by case status. Then, we compared the module scores between disease and control groups in the same brain region D. NotablTo confirm our transcriptome-based results at the protein level, we re-analyzed the tandem mass tag mass spectrometry (TMT-MS) dataset in AD . We inclThe module scores were correlated with amyloid plaque load (CERAD score), tau neurofibrillary tangle burden (Braak stage), cognitive function , and mass spectrometry molecular measurements  B. We obsWe associated our astrocyte modules with diagnostic classifications of AD C. In genAstrocytes intensively interact with surrounding cell types through signaling crosstalks, which are significantly perturbed in diseases. We explored the changes in intercellular signaling in different diseases with CellChat . We firsTo further investigate the critical disease-related cellular communication in which astrocytes are involved, we compared the significant changed ligand-receptor pairs among different disorders and target cell types D. SpecifWidespread molecular and functional diversity are recognized to exist in astrocytes across different brain regions and CNS disorders. The development of scRNA-seq enables a high-resolution view of astrocyte heterogeneity, and many studies demonstrated the molecular features of astrocytes in various diseases ,81,82,83+ astrocyte subpopulations located mainly in white matter. The new subtype highly expressed oligodendrocyte-associated genes. Although the existence of this subtype is controversial in many studies, we have confirmed the possibility of its expression of oligodendrocyte-associated genes from our aspect [We leverage the integrated dataset to better detect novel subpopulations and regulatory modules that could originally not be discovered in small datasets. For example, we identified cluster 3, OLIGO2r aspect ,11. Thror aspect ,48. We ar aspect ,51. SingSince we found that the conventional astrocyte composition analysis did not fully explain the occurrence of some diseases, we believe that explaining disease occurrence solely from the perspective of cell subtype abundance or functional modules is insufficient, and a combination of both is necessary. Some diseases may be related to multiple subpopulations, and it is unclear whether these subpopulations collectively contribute to disease occurrence. These factors cannot be explained by a single subpopulation alone, and we must examine them from the perspective of multi-subpopulation functional co-clustering modules. Therefore, we applied NMF analysis to construct seven functional specific co-expression gene modules. The M2 ECM and M4 stress modules showed evident signs of co-regulation, which generated a complex network of interactions. Furthermore, we validated that these two modules were significantly elevated in AD at both the transcriptome and protein levels. Notably, the M2 module was significantly elevated in the early stages of AD onset, which could furnish potential markers for AD early diagnosis. For example, M2 hub protein CD44 has been reported to have the potential as a fluid biomarker for the early diagnosis of AD by a recent proteomic study . MAFG, CD44, and DCLK1.These astrocyte transcriptomic modules recapitulate the core astrocyte features, which facilitate elucidating diseases mechanisms from a biological perspective. We identified aberration of the M3 synapse and M5 immunity in epilepsy, which are consistent with recent studies ,86. The Finally, we performed cellular communication analysis using integrated data and identified disease-specific signaling pathways that could contribute to disease pathogenesis. HD, PD, and Epi disorders correspond to highly disease-specific pathways. Particularly, the NRG3-ERBB4 signaling pathway from astrocytes to InN was significantly elevated in epilepsy. Therefore, we speculate that this signaling pathway of astrocytes may act as a protective function in epileptic pathology and NRG3 may be a new target for anticonvulsant drugs in epilepsy ,80. FutuIn summary, we integrated published datasets and identified novel and putative functional populations of astrocytes. Future functional studies are needed to evaluate these subpopulations and their potential effectiveness for modulation by therapies. How heterogeneity of astrocytes arises when subjected to different pathogenic stimuli is still an open question. We also hope that continued integration of our current data with future scRNA-seq and spatial datasets will add greater insights into solving these problems and making new discoveries.


Our recent study found that another purine derivative, paraxanthine, has neuroprotective effects similar to those of caffeine and UA. These purine derivatives can promote neuronal cysteine uptake through excitatory amino acid carrier protein 1 (EAAC1) to increase neuronal glutathione (GSH) levels in the brain. This review summarizes the GSH-mediated neuroprotective effects of purine derivatives. Considering the fact that GSH depletion is a manifestation in the brains of AD and PD patients, administration of purine derivatives may be a new therapeutic approach to prevent or delay the onset of these neurodegenerative diseases.Numerous basic studies have reported on the neuroprotective properties of several purine derivatives such as caffeine and uric acid (UA). Epidemiological studies have also shown the inverse association of appropriate caffeine intake or serum urate levels with neurodegenerative diseases such as Alzheimer disease (AD) and Parkinson’s disease (PD). The well-established neuroprotective mechanisms of caffeine and UA involve adenosine A A variety of purine derivatives are produced from purine nucleotides  by metabolic processes occurring mainly in the liver. For instance, adenine nucleotides are metabolized to produce ATP, ADP, AMP, adenosine, inosine, hypoxanthine, and xanthine, and finally uric acid (UA). Caffeine , a naturally occurring purine derivative, is metabolized into several dimethylxanthines including paraxanthine, theophylline, and theobromine, and finally into UA derivatives.2A receptor (A2AAR) antagonizing activity that is involved in neuroprotection [Each purine derivative has a particular physiological activity, and several purine derivatives such as adenosine, guanosine, caffeine, paraxanthine, theophylline, theobromine, and UA have been shown to possess neuroprotective activities ,6,7,8,9.otection . The neuotection ,11. The otection ,13,14,15otection ,17,18,19otection ,21. GSH otection ,23. Therotection ,21. Increasing GSH levels is a neuroprotective mechanism that increases antioxidant activity in neurons. It is widely accepted that neuroprotective processes can be classified into three major mechanisms: (i) suppressing excessive neuronal stimulation, (ii) maintaining antioxidant activity in neurons, and (iii) detoxification of xenobiotics . Intrace−  [The intracellular presence of the antioxidants such as ascorbate, UA, and α-tocopherol help to preserve neuronal GSH levels by doing the work of protecting cells against oxidative stress through other means ,41,42,43: 23657)   . A polymHigh serum UA levels cause some critical diseases such as gout, cardiovascular disease, hypertension, and renal disease , while iAlthough some conflicting results have been reported , a signiA strong correlation of plasma antioxidant levels with both mild cognitive impairment and AD has been observed ,86. FurtAs mentioned above, some studies have supported the potential role of hyperuricemia to prevent AD, while other studies have reported conflicting results or have failed to demonstrate any significant association ,96. AlthBy treatment with urate-lowering agents, patients with gout maintain lower serum UA levels over long periods of time. One might think that low serum UA levels would impair neurons by increasing oxidative stress in the brain. However, use of urate-lowering drugs  is not correlated with any increase in the risk of dementia . TreatmeSerum UA levels are maintained by food digestion, purine synthesis, metabolism, and purine excretion into urine. The net excretion of UA in urine is determined by the balance between UA re-absorption and secretion within the proximal tubule, and each process is mediated by its specific transporter . Since uUA is also produced during ATP metabolism through the following steps : (1) ATP2 mediate hypoxanthine oxidation to form xanthine and the end metabolite, UA, concomitant with super oxide anions (O2−). Therefore, the lack of XO in the brain may protect neurons from such oxidative stress. In vivo preclinical study has shown that UA protects hippocampal neurons after ischemia reperfusion in rats [In 1981, Granger et al. and McCord et al. ,108 hypo in rats .In terms of the mechanism by which UA in blood is involved in neuroprotection in the brain, Amaro et al.  argued t+/K+ ATPase and synaptic transmission [Other than nucleic acids, the typical intracellular purine derivative is ATP, which acts as an energy source for driving neuronal activity. Huge amounts of ATP  are produced in nervous tissues to maintain energy for Nasmission ,112,113.smission ,115,116 smission  and is asmission ,119,120 ATP is converted to ADP, AMP, adenosine, inosine, hypoxanthine, and xanthine and finally yields UA in the peripheral tissues . These p1 receptor (A1AR) and A2AAR  ,122,123.As described in the Introduction and illustrated in UA has antioxidant activity ,129,130 UA both acts as a scavenger of ROS and peroxynitrite  and prev2AAR receptor antagonism , while that of caffeine is AThere are numerous examples of UA’s direct neuroprotective roles. UA protects PC12 cells from 6-OHDA-induced cell injury by increasing levels of both GSH and superoxide dismutase (SOD) protein . UA treaUA also has an indirect neuroprotective activity by increasing GSH levels in astrocytes. UA treatment activates Nrf2 and leads to upregulation of gene expression of the GCLM, which enhances GSH production in astrocytes  Table 3Table 3. 2AAR antagonism is observed in the developing retina [Nrf2 activation is another indirect mechanism by which purine derivatives provide neuroprotection. Nrf2 activation was observed when caffeine was treated as a neuroprotective agent in lipopolysaccharide (LPS)-induced oxidative stress model mice  and cadmg retina , it is s1AR and A2AAR antagonisms, because an A2AAR antagonist, SCH58261, did not increase GSH levels in hippocampal slices [1AR  and UA (1 to 10 µM) provided neuroprotection by increasing GSH levels in the brain. The upregulation of GSH synthesis is mediated by increasing Cys uptake via EAAC1 in hippocampal slices . In addil slices , and becIn astrocytes, guanosine upregulates GCL, glutamine synthetase (GS), and GSH reductase (GR), resulting in elevated GSH levels ,137 Tab. The inc1AR and A2AAR , A2AAR (IC50 = 0.150 µM), adenosine A2B receptor  (IC50 = 15.4 µM), and adenosine A3 receptor  (IC50 = 6.5 µM), which are variously activated, depending on adenosine concentrations [1AR is the most abundant in the brain. Adenosine reduces neuronal excitability and firing rate via A1AR stimulation [1AR stimulation by adenosine induces neuroprotection in cultured neurons and ischemia model of rats [1AR stimulation in animal models of ischemia [1AR stimulation inhibits Ca2+ entry into the presynaptic terminal, resulting in a reduction in neurotransmitter release [2AAR stimulation in the brain induces neuronal excitability and synaptic transmission [2AAR antagonists such as KW-6002 (istradefylline), which is xanthine-based compound, protect nigral dopaminergic cells from damage induced by 6-OHDA in rats or by MPTP in mice [2AAR lowered the dopaminergic neurotoxicity in PD model animals [1AR agonism and A2AAR antagonism [Adenosine is another purine that has neuroprotective activities. Since the 1940s, adenosine has been used clinically for cardiac protection and vasodilation . Adenositrations ,148,149 IC50 = 0. µM, adenmulation ,152. A1Aischemia . A1AR st release ,155,156  release ,158. In smission ,159. Spe in mice . Genetic animals . These r2AAR receptor, as well as its previously described category (ii) role of increasing intracellular GSH levels via Cys uptake. Methylxanthines such as caffeine and theophylline are known for their bronchoprotective effects. The molecular mechanisms include adenosine receptor antagonism  and phosphodiesterase inhibition  [1AR antagonism is involved in the stimulus effects of caffeine [2AAR antagonism [1AR and A2AAR [1AR or the A2AAR, suggesting that this effect is independent of AR activity. It is accepted that the neuroprotective activities of caffeine are mediated by A2AAR inhibition [Caffeine is another purine that offers category (i) neuroprotection through the A 400 µM)  (Table 2 400 µM) , it is e 400 µM) ,162. A1Acaffeine . Of thestagonism , and thend A2AAR ,166. Thehibition . Caffeinhibition . MechaniThere are a few other, less pervasive mechanisms responsible for neuroprotective activities of purines. UA promotes pro-survival pathways such as activating PI3K/Akt, and MEK/ERK in the brain. For example, UA treatment (200 µM UA) prevents dopamine (DA) neuron loss and behavioral deficits in 6-OHDA treated rat by recovery of Akt/GSK3β signaling  Table 3Table 3. Ya et al.  have shoParaxanthine at a concentration of 800 µM has a neuroprotective activity for dopaminergic neurons . Its neuATP itself has no neuroprotective activity corresponding to categories (i) and (ii) in 2AAR, increasing GSH levels via Cys uptake may play an important role in the neuroprotective activities of purine derivatives. As described above, the neuroprotective mechanisms of purine derivatives are mainly due to adenosine receptor modulation and upregulation of the synthesis of proteins that are involved in increasing antioxidative activity or activating pro-survival pathways in cells. Many receptor activities develop tolerance to their antagonists , i.e., cSeveral purine derivatives exhibit neuroprotective activity mediated by increasing neuronal GSH levels, which can be due to not only induction of GSH synthesis-related enzymes, but also the upregulation of GSH levels explained by promoting EAAC1-mediated Cys uptake, which is a commonly seen after treatment with caffeine, UA, or paraxanthine.The upregulation of GSH levels may enhance antioxidative activity in neurons and appears to be effective in preventing the incidence and progression of neurodegenerative disease. Further studies to investigate how purine derivatives increase GSH levels by promoting EAAC1-mediated Cys uptake would provide valuable insights into neuroprotection by purine derivative treatment. These studies of purine derivative biochemistry could lead to novel preventions and treatments for the growing burden of neurodegenerative disease in aging societies worldwide.