Cardiorespiratory fitness (CRF) and physical activity (PA) are well-established predictors of morbidity and all-cause mortality. However, CRF is not routinely measured and PA not routinely prescribed as part of standard healthcare. The American Heart Association (AHA) recently presented a scientific case for the inclusion of CRF as a clinical vital sign based on epidemiological and clinical observation. Here, we leverage genetic data in the UK Biobank (UKB) to strengthen the case for CRF as a vital sign and make a case for the prescription of PA.We derived two CRF measures from the heart rate data collected during a submaximal cycle ramp test: CRF-vo2max, an estimate of the participants' maximum volume of oxygen uptake, per kilogram of body weight, per minute; and CRF-slope, an estimate of the rate of increase of heart rate during exercise. Average PA over a 7-day period was derived from a wrist-worn activity tracker. After quality control, 70,783 participants had data on the two derived CRF measures, and 89,683 had PA data. We performed genome-wide association study (GWAS) analyses by sex, and post-GWAS techniques to understand genetic architecture of the traits and prioritise functional genes for follow-up.We found strong evidence that genetic variants associated with CRF and PA influenced genetic expression in a relatively small set of genes in the heart, artery, lung, skeletal muscle and adipose tissue. These functionally relevant genes were enriched among genes known to be associated with coronary artery disease (CAD), type 2 diabetes (T2D) and Alzheimer’s disease (three of the top 10 causes of death in high-income countries) as well as Parkinson’s disease, pulmonary fibrosis, and blood pressure, heart rate, and respiratory phenotypes. Genetic variation associated with lower CRF and PA was also correlated with several disease risk factors ; a typical T2D profile ; increased risk for CAD and T2D; and a shorter lifespan.Genetics supports three decades of evidence for the inclusion of CRF as a clinical vital sign. Given the genetic, clinical and epidemiological evidence linking CRF and PA to increased morbidity and mortality, regular measurement of CRF as a marker of health and routine prescription of PA could be a prudent strategy to support public health.The online version contains supplementary material available at 10.1186/s13073-021-00994-9. Cardiorespiratory fitness (CRF) and physical activity (PA) are well-established predictors of morbidity and mortality , 2. CRF Sedentary lifestyles are a significant public health problem. For example, physical inactivity and poor diet were the second leading modifiable, behavioural cause of death in the US in 2000 and are expected to overtake tobacco to become the leading cause . Worldwih2=0.25–0.65 ), the most prevalent of the top 10 global causes of death. Diagnoses were potentially received both before and after the cycle ramp test and the week in which participants wore the physical activity tracker, and we made no distinction between incident and prevalent disease. This analysis was performed separately in males and females, since there are differences in both disease prevalence and in phenotype distribution by sex.The top 10 global causes of death in high-income countries in 2016, as reported by the World Health Organization (WHO), are shown in Additional file The full UKB sample was genotyped on one of two custom Applied Biosystems genotyping arrays: 49,950 on the UK Biobank Lung Exome Variant Evaluation (UK BiLEVE) Array, and 438,427 on the UK Biobank Axiom Array. The genetic data underwent standard GWAS QC and were imputed using IMPUTE4 to three reference panels by the UKB: the Haplotype Reference Consortium (HRC) and the merged UK10K and 1000 Genomes phase 3 reference panels. General genotyping considerations, raw genotype data QC and genetic imputation procedures in the UKB are described in detail elsewhere . Details2, sex , array, centre and the first 10 principal components calculated on subsamples with data on the trait of interest. Additionally, for CRF phenotypes, we included the covariates risk category (corresponding to CRF test protocol) and number of trend entries (number of observations). Full details of GWAS QC and association parameters, alternative models including BMI, and alcohol and smoking as covariates, and post-GWAS analyses including transcriptome-wide association (TWAS) and colocalisation, gene-set enrichment and genetic correlation, are included in the supplementary methods. We used the standard p=5 × 10-8 as our genome-wide significant threshold, a conservative Bonferroni-corrected p value for TWAS significance correcting for all genes tested in all tissues , and a false-discovery rate (FDR) correction for gene-set enrichment and genetic correlation accepting FDR-adjusted p values<0.05 as significant.From UKB-supplied sample QC information, we excluded gender mismatches, samples with putative sex chromosome aneuploidy or excess relatives, and heterozygosity and missingness outliers. We performed linear mixed model association on the residual of each trait regressed on the covariates age, ageFor data analysis and figure creation, we used both R  and PythTable S4). Correlations between CRF and PA phenotypes, and age and BMI were all modest and negative (r=−0.02–−0.27), except for CRF-slope and age which had a small positive correlation in females  . Combined-sex sample correlations between CRF-vo2max and CRF-slope, and SES were not significant; the PA and SES, and sex-specific correlations for all three traits were significant but small (| r |<=0.03). In the combined-sex sample, phenotypic correlations between our three traits were in the directions expected. CRF-vo2max and PA had a modest positive correlation . CRF slope and PA were not significantly correlated . The two fitness phenotypes, CRF-vo2max and CRF-slope were negatively correlated .Descriptive statistics for the CRF and PA phenotypes showed higher values for CRF-vo2max in males and higher CRF-slope and PA in females  and had a near zero correlation with CRF-slope .As a proxy for true resting heart rate, heart rate at the beginning of the exercise phase of the cycle ramp test was negatively correlated with CRF-vo2max (Table S5), which contributed little to trait variance in linear regression models of all three phenotypes . We chose not to adjust for the effects of smoking and drinking status in order to obtain unbiased estimates of the genetic factors on CRF and PA. There is evidence that correcting for heritable covariates can lead to biassed estimates of the influence of genetic factors [Smoking and alcohol drinking status are sometimes considered confounding factors in analyses of CRF and PA. In our data, there were small differences in mean CRF-vo2max, CRF-slope and PA by smoking and drinking status , With increasing levels of CRF-vo2max, there were reductions in diagnosis frequency for ischaemic heart disease (CAD) , stroke , chronic obstructive pulmonary (COPD) , lower respiratory tract infection (LRTI) , diabetes mellitus  and kidney diseases . The largest differences were observed for CAD and diabetes, with the starkest reductions for all diseases often seen within the lowest 4 deciles of CRF-vo2max.CRF-slope showed reduction in disease for CAD  and diabetes . The relationships with other disease diagnoses were noisier.Disease diagnosis by level of PA showed similar patterns to CRF-vo2max, with a continued decile-to-decile reduction in disease across the range of PA from least to most physically active. With increasing PA we observed reductions in CAD , stroke , COPD , LRTI , diabetes mellitus  and kidney diseases . Like CRF-vo2max, the greatest reductions were seen in CAD and diabetes mellitus.Figures S3–S37 are the regional plots for each sex-combined GWAS signal peak and sex-specific GWAS signal peaks where there was evidence of heterogeneity.Genome-wide association studies were performed for CRF-vo2max, CRF-slope and PA with results for male, female, and the combined sample Fig. . An annor2=0.78 in 1000 Genomes EUR .We found twelve significant SNPs for CRF-vo2max and eight independent SNPs for CRF-slope, in the combined sample Table . For PA,r2=0.79 with rs6801957, chr3), rs111299422 (chr5), rs1006545  and rs41317306 (chr14), however showed no evidence for association in females (p=5.1 × 10-1), or the combined sample (p=1.6 × 10-4), and significant effect size heterogeneity . However, the regional plot for rs41317306 did not show support from correlated SNPs For CRF-vo2max, we found four male genome-wide significant SNPs, three of which showed no evidence of heterogeneity and were associated (directly or indirectly through LD) in the combined sample: rs9809798 . There were two male-specific SNPs: rs1006545 (chr10) was indirectly associated in the combined sample (r2=0.97 with 10:102554618_AT_A) and showed no evidence of heterogeneity. rs1741294 (chr20) showed no evidence of association in females (p=6.0 × 10-1) or the combined sample (p=4.2 × 10-4), and significant effect size heterogeneity .For CRF-slope, the one female signal rs111299422 (chr5) was significant in the combined sample, but showed evidence of heterogeneity , and showed no evidence of heterogeneity. The two female signals on chromosome 16 signals rs75986475 and rs13329850 were uncorrelated (r2=0.0005), showed significant heterogeneity , and no evidence of association in males  or the combined sample . The male-specific SNP identified in PA, rs78661713 (chr2), showed no evidence for association in females (p=6.1 × 10-1) or the combined sample (p=5.9 × 10-4), and significant heterogeneity .For PA, there were four SNPs associated in the female sample, two of which were associated in the combined sample, rs62055696 (chr17) and 17:44262581_A_C  in the aorta, heart , lung and skeletal muscle; MAPT (microtubule-associated protein tau) in the brain and lung; LRRC37A2 (leucine-rich repeat-containing 37A2) in and LRRC37A (leucine-rich repeat-containing 37A) in the adipose , artery , heart , and skeletal muscle and CRHR1 (corticotropin-releasing hormone receptor 1) in the adipose  and skeletal muscle. Notable GWAS catalogue associations for this set of genes include disease , lung fillation , peak exillation ), heart illation , cardiacillation ), and blillation , haemoglillation , haematoillation  and red illation ).ACHE (acetylcholinesterase), implicated by rs4582488 (52.87kb upstream), was significantly associated with expression in the adipose , artery , and heart  tissue. Each of these tissue associations had a high probability of a shared SNP driving both the CRF-slope association and expression (PP4 range=0.79–0.84). ACHE has notable reported associations with resting heart rate [Positional candidate art rate , electroart rate , diastolart rate , and diaart rate .TRIP6 (thyroid hormone receptor interactor 6) with a high probability of a shared SNP driving both CRF-slope association and expression in the whole blood, and PINX1 (PIN2 (TERF1) interacting telomerase inhibitor [ERI1 (exoribonuclease 1) had significant expression in the lung, with strong evidence the same variant drive expression and the CRF-slope association (PP4 range=0.71–0.98). TRIP6, PINX1, and ERI1 have notably been associated with blood pressure [Three functional candidates were also significant in the gene-based association: pressure , triglycpressure , HDL chopressure , diabetepressure , and athpressure .SRRT  in the lung and heart  tissue, RP11-62H7.2 in the heart , FAM86B3P (family with sequence similarity 86 member B3) in the skeletal muscle, FAM85B (family with sequence similarity 85 member B) in the lung and coronary artery, and AF131215.2 in the artery  (PP4 range=0.74–0.92). SRRT and FAM86B3P are notably associated with heart rate response to exercise and recovery [Additional TWAS significant functional candidates with a high probability of the GWAS variant driving expression included recovery , electrorecovery , diabeterecovery , and oberecovery .L3MBTL2 on chromosome 22. The number of loci and the best candidates should be interpreted with caution as this is a gene-dense region with many copy number variants, and all the chromosome 17 candidates were within or near the large 17q21.3 inversion polymorphism [All PA functional candidates were on chromosome 17, except morphism , 45. NonPLEKHM1 (plekstrin homology domain containing M1) in the lung, skeletal muscle and peripheral blood with low probability of the GWAS variant driving expression (PP4 range=0.30–0.33); NSF  in the coronary artery (PP4=0.54); MAPT in the subcutaneous adipose (PP4=0.91), artery , and brain, left ventricle, lung, adipose (PP4 range=0.93–0.99); LRRC37A2 in the adipose , artery , heart , lung ; KANSL1 (KAT8 Regulatory NSL Complex Subunit 1) in the skeletal muscle (PP4=0.92); CRHR1 in adipose  and skeletal muscle (PP4≥0.98); ARL17A (ADP ribosylation factor like GTPase 17A) with high probability of a shared variant driving PA association and gene expression in left ventricle (PP4=0.88) and skeletal muscle (PP4=0.97); ARHGAP27 (rho GTPase activating protein 27) in visceral omentum (PP4=0.92); and L3MBTL2 (L3MBTL histone methyl-lysine binding protein 2) in subcutaneous adipose (PP4=0.67).Ten significant genes from the gene-based association analysis were identified as functional candidates: Previous notable associations with these ten functional candidates include disease , CRF-slope (10 genes) and PA , as well as female PA (22 genes: 21 with recognised Ensembl ID). TWAS analyses of the other sex-specific traits did not yield sufficient gene associations to perform enrichment analyses. Enrichment of TWAS-identified functional candidates in gene sets defined by genes identified in previous GWASs, reported in the GWAS catalogue, are shown in Additional file 6 genes, p range=6.9 × 10-3–2.0 × 10-5), as well as lung function  and blood phenotypes . Enrichment for genes implicated in Alzheimer’s and Parkinson’s diseases is noteworthy given there is some evidence of an underlying insulin resistance pathology.Functional candidates for CRF-vo2max were enriched in GWAS gene sets for multiple diseases including multiple system atrophy, Alzheimer’s disease, Parkinson’s disease, idiopathic pulmonary fibrosis and primary biliary cirrhosis (FDR-adjusted PAI-1 (plasminogen activator inhibitor-1: FDR-adjusted p=5.0 × 10-6), heart rate response to exercise and recovery (FDR-adjusted p=1.2 × 10-5) and resting heart rate (FDR-adjusted p=1.4 × 10-3).CRF-slope functional genes were enriched in GWAS gene sets for the thrombosis and atherosclerosis risk factor p range=2.7 × 10-2–2.0 × 10-8), and mental health traits . Also noteworthy was enrichment of female PA-associated functional candidates in FEV1 (FDR-adjusted p=4.1 × 10-4) and FVC (FDR-adjusted p=3.1 × 10-3) given their use in the diagnosis of obstructive and restrictive pulmonary diseases.GWAS gene sets enriched for PA functional candidates included Parkinson’s disease, idiopathic pulmonary fibrosis, Alzheimer’s disease, multiple system atrophy and primary biliary cirrhosis , suggesting genetic variants that contribute to greater fitness are also associated with a lower rate of increase of heart rate during exercise. PA had a moderate positive correlation with CRF-vo2max , and a modest negative correlation with CRF slope . These two genetic correlations suggest that higher levels of activity have a shared genetic contribution with greater fitness and with a lower rate of increase of heart rate during exercise. The within-sex between-phenotype genetic correlations were similar to those for the sex-combined results  and higher fasting insulin , impaired beta-cell function , hyperglycaemia  and hypertriglyceridemia . In the PA combined sample, as well as genetic correlations with fasting insulin , insulin resistance  and triglycerides , we observed a positive correlation with HDL cholesterol  and a direct negative correlation with T2D . Genetic variation associated with lower levels of physical activity was associated with increased risk of T2D.Genetic variation associated with lower CRF-vo2max was correlated with features of the typical type-2 diabetes (T2D) profile: higher insulin resistance . Like T2D, genetic variation associated with lower CRF-vo2max and lower PA was associated with increased risk for coronary artery disease. Additional file Both CRF-vo2max and PA were negatively correlated with coronary artery disease , such that genetic variation associated with greater CRF-vo2max was associated with lower levels of the GlycA biomarker. Elevated blood concentrations of GlycA are a strong predictor for long-term risk of morbidity and mortality from diverse diseases [CRF-vo2max was significantly genetically correlated with glycoprotein acetyls, GlycA . Genetic variation associated with greater CRF-vo2max and PA was associated with higher levels of HDL. PA was also significantly correlated with acetate  and apolipoprotein A-I —the major protein component of HDL which enables fat efflux from within cells for transport to the liver for excretion. Additional file For PA, we observed positive FDR-significant genetic correlations with several related HDL metabolites. The largest HDL metabolite correlation with PA was total cholesterol in medium HDL , and for BMI-adjusted leptin  , birth weight  and own birth weight . All other FDR-significant anthropometric correlations were negative : body fat, waist circumference, measures of obesity, BMI, waist-to-hip ratio and hip circumference. CRF-slope had FDR-significant negative genetic correlations with childhood obesity  and childhood height , BMI , birth weight  and own birth weight . For PA, all FDR-significant body measures including height were negative : body fat, waist circumference, measures of obesity, BMI, waist-to-hip ratio and hip circumference, and height. Genetic variation associated with greater CRF-vo2max and higher levels of PA was associated with lower body fat, BMI, obesity, and waist and hip circumferences. Additional file CRF-vo2max was positively genetically correlated with height , forced expiratory volume in 1 s  and peak expiratory flow . CRF-slope was negatively correlated with FVC  and peak expiratory flow . For PA, we found modest positive genetic correlations with FVC  and FEV1 . Genetic variation associated with both fitness and activity is associated with improved lung function.CRF-vo2max was positively associated with lung function: forced vital capacity . Genetic correlation associated with greater acceleration of heart rate during exercise was associated a lower resting heart rate. Additional file Only CRF slope was genetically correlated with resting heart rate rg=−0.3, se=0.07rg=0.27, se=0.07). A genotype associated with greater levels of activity associated with smoking cessation.We observed a positive genetic correlation between PA and former vs current smoking status  and parents’ ages at death . FDR-significant genetic correlations with smoking behaviour, cancer and longevity reported above are highlighted in Additional file Finally, we found that genetic variation associated with greater CRF-vo2max and PA was also significantly associated with longevity, as measured by father’s age at death  and age at birth of first child . CRF-slope had a corresponding negative genetic correlation with years of schooling . FDR-significant genetic correlations with education and reproductive phenotypes are highlighted in Additional file CRF-vo2max and PA were positively correlated with years of schooling . The two derived fitness traits, CRF-vo2max (vo2max) and CRF-slope (rate of increase of heart rate during exercise), were negatively correlated, phenotypically and genetically. Greater levels of fitness corresponded to a slower rate of increase of heart rate during exercise. CRF-vo2max was positively correlated with PA; greater fitness was associated with higher levels of physical activity.We found that higher levels of CRF-vo2max and PA were associated with better health outcomes in a dose-dependent way. The frequency of CAD and diabetes mellitus diagnosis was substantially lower among individuals with higher levels of CRF-vo2max and PA, even after controlling for BMI and SES. In this large UK population-based sample, we find the same associations between PA and CRF-vo2max observed in epidemiological and clinical studies of coronary heart disease and cardiovascular events  and T2D WNT3, MAPT, LRRC37A2, LRRC37A, CRHR1). These five genes have previously been associated with GWASs for atrial fibrillation [Our functional genetic analyses identified genes associated with CRF and PA, with known associations in disease and clinical vital sign-related phenotypes. For CRF-vo2max, we found functional candidates associated with expression in the heart, artery, lung, brain, adipose and skeletal muscle. All functional candidates were within 17q21.31, with five genes also identified in the gene-based association analysis , heart illation , cardiacillation  and blooillation , haemoglillation , haematoillation  and red illation ). SubsetACHE within 7q21.1, also identified as a positional candidate 52.87kb upstream of rs4582488, was significantly associated with expression in adipose, artery and heart tissue and has previously been associated with resting heart rate [TRIP6, PINX1 and ERI1) were significantly associated with expression change in the whole blood and lung and have previously been associated with blood pressure [SRRT, RP11-62H7.2, FAM86B3P, FAM85B and AF131215.2) were associated with expression in the lung, heart, skeletal muscle and coronary and tibial artery and have been associated in previous GWASs with heart rate response to exercise and recovery [PAI-1, heart rate response to exercise and recovery, and resting heart rate.For CRF slope, functional candidates fell within one of two cytogenic regions, 7q21.1 and 8p23.1. art rate , electroart rate , diastolart rate  and diabart rate . The genpressure , triglycpressure , HDL chopressure , diabetepressure  and athepressure . Other precovery , electrorecovery , diabeterecovery  and obesrecovery . We founMAPT, LRRC37A2, LRRC37A, KANSL1, CRHR1, ARL17A and ARHGAP27) and 22q13.2 (L3MBTL2) with a high probability of the same variant driving PA association and expression in the adipose, brain, heart, lung and muscle; have previously been associated with disease  anthrop disease ), blood  disease , red blo disease , eosinop disease ), lung f disease , FEV1 [5 disease ), and he disease , cardiac disease ). We fouGenetic correlations complement epidemiological and clinical observations. Because inherited genetic risk cannot be due to confounding, genetic correlations limit the number of potential confounders linked through genotype. The shared genetic architecture of both CRF-vo2max and PA clearly implicated the lifestyle-related chronic diseases obesity, T2D and CAD, and their body composition and metabolic risk factors. CRF-vo2max and PA were negatively genetically correlated with obesity, body fat, body mass index and waist-to-hip circumference, triglycerides, the satiety hormone leptin, fasting insulin, insulin resistance, and T2D (significant only for PA), and CAD. These negative genetic correlations corroborate the epidemiological associations, previous research findings and clinical observations of CRF and PA and body composition , 78, lipCRF-vo2max was negatively genetically correlated with blood levels of glycoprotein acetyls (GlycA). GlycA is associated with both acute and chronic inflammation and is a strong predictor for long-term risk of morbidity and mortality from a wide range of diseases .Additionally, CRF-vo2max and PA had positive genetic correlations with age at birth of first child and level of education, respectively. a correlate and component of SES. CRF slope showed a corresponding negative correlation with years of education. This is consistent with population-based observations of a socioeconomic gradient in PA and CRF: people living in higher socioeconomic status areas tend to be more active and more fit and from a public health perspective particularly important as ‘reduced PA and fitness levels may contribute to social inequalities in health’ .N=24,925 compared to body fat N=100,716), and many metabolite correlations failed to pass the multiple testing threshold. As well as this sample size (or power of each discovery GWAS) limitation, genetic correlations cannot resolve reverse causation. Another limitation is correlated lifestyle risk factors not considered, in particular eating behaviours and nutrition. Eating habits, diet and exercise behaviours tend to go hand in hand, in large part influenced by the environment, e.g. sedentary time spent watching television and eating high-fat foods [We note several limitations. The gold standard for CRF measurement involves exercise to volitional exhaustion with measurement of gas exchange . A submaximal CRF test with extrapolation of VO2max scores is however a much more practical solution, especially for the collection of a large sample in the age ranges studied. The objective PA monitoring only captured a 7-day window, effectively a snapshot of participant behaviour, and we do not know if this reflects their habitual behaviour. We also used raw acceleration and did not attempt to tease apart PA intensity, recognised as important for health outcomes. All genetic associations, especially the sex-specific SNPs identified, will require replication in an independent sample. In our genetic correlation analyses, some of the largest correlations were observed with blood metabolites but these were based on relatively small GWAS reference samples , Results , Tables S1 – S9, and Figures S1 – S52.


KTI12, is closely associated with plant biomass and yield. KTI12 is involved in developmental processes of most organs, including the leaf, root, flower, and seed, through regulating cell division and differentiation. Previous work has shown that in upland cotton (Gossypium hirsutum), GhKTI12 regulates plant height, flowering, and tolerance to salt and drought stress. However, little is known about the molecular regulation mechanism of GhKTI12 in plant developmental processes. In this study, we identified the main GhKTI12 (Gh_D02G144400) gene and transformed it into tobacco (Nicotonia tabacum cv NC89). From seven transgenic lines, we obtained three  with high expression of GhKTI12; compared with wild type plants, these three lines exhibited larger plant size, later flowering, and higher seed yield. Microscopic observation revealed that the number of leaf epidermal cells and stem parenchyma cells was increased by ~55%. Biochemical analysis showed that chlorophyll content and starch accumulation were significantly increased in younger leaves at the top canopy of transgenic plants, which may contribute to improved photosynthetic rate and, in turn, increased seed yield. To understand the molecular mechanism of GhKTI12 in transgenic plants development, two lines (OE6 and OE8) with higher expression levels of GhKTI12 were used as representative plants to conduct RNA-seq analysis. Through transcriptome analysis of the plant’s shoot apical meristematic tissue of these two lines, we identified 518 upregulated genes and 406 downregulated genes common to both overexpression lines. A large number of cellular component genes associated with cell division and differentiation, such as RD21, TET8, KTN80, AOX1, AOX2, CP1, and KIC, were found to be upregulated, and genes showing the most downregulation included MADS-box genes related to flowering time, such as MADS6, AP1, AP3, AGL8, AGL6, SEP1, and SEP2. Downregulation of these genes caused delayed flowering time and longer vegetative stage during development. Combined with the upregulation of the yield-related gene RD21, the GhKTI12 transgenic plants could produce a higher seed yield. We here show that the overexpression of GhKTI12 could positively improve key agronomic traits in tobacco by regulating cell proliferation, photosynthesis, and organ development, and suggest that homologs of GhKTI12 may also be important in the genetic improvement of other crop plants.Crop molecular breeding primarily focuses on increasing the trait of plant yield. An elongator-associated protein,  KTI (Killer Toxin Insensitive or Kluyveromyces lactis Toxin Insensitive) gene encodes an elongator-associated protein. Three KTI genes  were initially isolated from Saccharomyces cerevisiae [KTI12 is a highly conserved ATPase, and is necessary for the tRNA-modification activity of the eukaryotic Elongator complex [KTI12 homologs have been identified in many plants, such as Arabidopsis thaliana, Oryza sativa, Zea mays, Brassica napus, Populus [The revisiae . KTI12 i complex . KTI12 h Populus .Yeast KTI12 and plant orthologs contain an N-terminal P-loop motif  and a puAtKTI12 and AtELP1 in Arabidopsis were shown to reduce organ growth and early flowering, respectively [Plant KTI12 homologs are localized to the nucleus  and reguectively .DEFORMED ROOTS AND LEAVES1 (DRL1) is a KTI12 homolog that can modulate cell division and differentiation to regulate meristem activity and organ growth in Arabidopsis [AtDRL1/KTI12 mutants in Arabidopsis have defective shoot and root meristems [KTI12 with Arabidopsis and rice drl1 mutant showed rescued growth retardation [ELP mutants in Arabidopsis show pleiotropic phenotypes including narrow leaves, enlargement of hypocotyl, retarded primary root growth, decreased seed germination, delayed flowering time, and reduced apical dominance [bidopsis ,8. AtDRLeristems ,15. Compominance ,9,16,17.Nicotiana tabacum L.) is an economically important plant, and the variety NC89 has been widely used as a model for gene functional analysis. In a previous study, we screened functional genes related to plant architecture, abiotic stress tolerance, and yield in cotton using RNA-seq data and the virus-induced gene silencing (VIGS) method. We found that KTI12 affects plant height, flowering, and tolerance to salt and drought stress in upland cotton. There are two homologous genes, Gh_A03G123100 and Gh_D02G144400, in the cotton genome. From gene expression profiles and stress-induced characteristics, we found that Gh_D02G144400 is likely the gene primarily responsible for KTI12 function. However, very few studies have functionally characterized the GhKTI12 gene. In this study, we transformed GhKTI12 (Gh_D02G144400) into the tobacco variety NC89. Transgenic tobacco plants with high expression of GhKTI12 were identified and used to analyze gene function based on phenotypes. The results of this study provide a better understanding of KTI12 gene function in key plant traits and lay an experimental foundation for genetic crop improvement.Tobacco  medium Petri plates and incubated at 25 °C in a growth chamber with a 16 h photoperiod for 15 days. Precultured leaves were used as explants for Agrobacterium tumefaciens-mediated genetic transformation .GhKTI12 into the empty vector pCAMBIA2300-35S between KpnI and SmaI sites. The pCAMBIA2300-35S plasmid was provided by Biotechnology Research Institute, Chinese Academy of Agricultural Sciences in Beijing, China. The overexpression vector was transformed into Agrobacterium strain GV3101. Transgenic tobacco plants were created by the Agrobacterium-mediated leaf disc method [GhKTI12 gene. The PCR program was as follows: pre-denaturation at 95 °C for 3 min; 30 cycles of amplification ; followed by 5 min at 72 °C. T2 homozygous lines were generated to conduct further experiments.The overexpression vector pCAMBIA2300-35S-GhKTI12 was constructed by inserting the coding sequence of c method . After o−ΔΔCT method was used to calculate relative expression level [NtACT7 was used as the internal reference gene. Primers are shown in For transcript expression analysis, RNA from different tissue samples was extracted using a FastPure Plant Total RNA Isolation Kit . First-strand cDNA synthesis was conducted with HiScript II Q RT SuperMix for qPCR (+gDNA wiper) kit , following the manufacturer’s instructions. The 2on level , and NtAMorphological traits of transgenic and WT plants were recorded for the T2 generation including leaf size, plant height, number of leaves, number of internodes, flowering time, capsule number, and seed yield at flowering time. The fresh and dry weight of vegetative tissues  were measured. Transgenic plants were separately harvested and dried at 70 °C after harvesting.g for 10 min. Absorbance of the supernatant was measured at 649 and 665 nm with a Spark plate reader .Chlorophyll content in transgenic and WT plants was determined as described by Parry (2014) . BrieflyLeaf tissue and stem internode samples were collected from 45- to 60-day-old GhKTI12 transgenic and WT plants, then fixed in a solution of 3:1 ethanol: acetic anhydride with a drop of Tween-20 for 1 h . SamplesGhKTI12 overexpressing lines at the juvenile vegetative stage (approximately 60 days after sowing). A total of 18 leaf samples of three individual plant were selected from each transgenic line. Starch and sucrose content were measured using the β-Amylase Assay Kit and Sucrose Assay Kit  according to the manufacturer protocols.For sucrose and starch content analysis, leaves were harvested from different positions  of three plants for the WT and each of the p-value < 0.05 were considered significant. The expression profile of DEGs were visualized with HemI  [Total RNA was extracted from the shoot apical meristem of 40-day-old plants with three samples for each line  using the RNeasy Plant Mini Kit . Libraries for RNA-seq analysis were constructed from 3 mg of total RNA per sample using the RNA Library Prep Kit for Illumina . Constructed libraries used Illumina high throughput sequencing platform NovaSeq 6000 for sequencing. Differentially expressed genes (DEGs) were identified and gene ontology (GO) enrichment analysis was performed using TBtools  and agrir, v1.0) .p-value. Differences between transgenic lines and WT were considered if p < 0.05. Significantly, differences between transgenic lines and WT were considered if p < 0.01.Data were analyzed using SPSS 20.0. Means were compared within transgenic and WT groups using Tukey’s test at the 5% and 1% level. All of the morphological data for T2 transgenic progeny are presented as the mean ± standard error and GhKTI12 homologs in upland cotton, and Gh_D02G144400 is the main functional gene. The full-length coding sequence of GhKTI12 (Gh_D02G144400) is 912 bp and encodes a protein of 303 amino acids in length with a molecular weight of 34 kDa and an isoelectric point of 8.9 , rice ( Oryza sativa subsp. japonica and indica), maize (Zea mays),upland cotton, diploid cotton (Gossypium raimondii), island cotton (Gossypium barbadense), Gossypium arboretum, tobacco, arabica (Coffea arabica), white poplar , Leprosy tree (Jatropha curcas), sesame (Sesamum indicum), potato (Solanum tuberosum), and rape (Brassica campestris)  a. The phGhKTI12 overexpression vector . The average leaf length in transgenic plants was 4.83 cm, 4.79 cm, and 4.60 cm at the 5th, 8th and 12th position, respectively. This was ~36%, 41%, and 59% higher than the corresponding leaves from WT plants a. AlthouKTI12 played a positive role in regulating leaf length by increasing cell proliferation rather than cell expansion.Plant organ development size was primarily determined by cell proliferation and cell expansion. To analyze the mechanism of leaf size changes in transgenic plants at the cytological level, we observed cell number and size in the third leaf of 45- to 60-day-old plants under the microscope. The average number of epidermis cells was significantly increased and average cell size was decreased in transgenic plants compared to the WT c; cell nGhKTI12 is involved in the positive regulation of leaf epidermal cell division and proliferation for leaf area enlargement in transgenic tobacco plants.Taken together, our results support that GhKTI12 could promote root development to increase root volume.All of the OE plants were larger than WT plants at the reproductive stage , particuOE-5, OE-6 and OE-8 also produced more flowers and capsules in the reproductive stage. The average capsule number per ten plants in OE-5, OE-6 and OE-8 was 114, 128 and 142, which contributed to an increase of 35%, 40% and 44% in net seed yield, respectively d,e, compGhKTI12 could enhance chlorophyll production during leaf development in transgenic tobacco plants.In OE plants, leaves appeared greener from the middle to the 10th leaf upper positions than those of WT plants, but had a comparable yellow color in the 6th leaf lower position. Chlorophyll contents were increased approximately 12% and 35% at the 6th and 10th position, respectively, in transgenic plants a. ChloroGhKTI12 could increase chlorophyll content and carbohydrate accumulation, promoting plant development and high yield.In order to understand the mechanism of higher yield and the accumulation of photosynthetic products in transgenic plants, sucrose and starch contents were analyzed in the 5th–10th leaves of 60-day-old OE and WT plants. The leaf sucrose content was gradually increased in the 5th to 8th leaf of transgenic plants, and declined to 13 mg/g fresh weight (FW) in transgenic younger leaves at the 9th position, approximately 16% lower than in the 9th leaf of WT plants b. ConverKTI12 interacts with elongator complex to regulate gene expression. Previous results revealed that the overexpression of GhKTI12 could positively affect plant development. However, there was no significant difference in morphology traits between different transgenic lines . To explore the molecular mechanism of global gene expression changes in transgenic plants, the OE-6 and OE-8 with most high expression of GhKTI12 gene were selected to do transcriptome analysis. The transcriptome analysis samples were shoot apical meristematic tissue (SAM) of 60-day-old plants which considered to be the tissue with the most vigorous cell division and differentiation. We identified 11,168 differentially expressed genes (DEGs) from the RNA-seq data comparison of OE-6 and OE-8 with WT plants. Among those DEGs, 2677 (24%) were identified as core DEGs  to gain insight into core DEG functions. The 518 core upregulated genes were enriched in 28 GO functional groups; these included six cellular components, 11 biological processes, and 11 molecular functions . The terp-value < 0.05, FDR < 0.001) respiratory chain (GO:0070469), lysosome (GO:0005764), endoplasmic reticulum lumen (GO:0005788), lytic vacuole (GO:0000323), and “microtubule organizing center part” (GO:0044450). Significantly enriched biological processes were related to immune and heat response, namely regulation of systemic acquired resistance (GO:0010112) and response to heat (GO:0009408). Significantly enriched molecular functions were unfolded protein binding (GO:0051082), glutathione transferase activity (GO:0004364), and α-L-fucosidase activity (GO:0004560) (Enriched cellular components included the highly significant (0004560) .AOX) genes , four cytochrome c-dependent genes , two ATPase inhibitor proteins (At2g27730 and F1F0-ATPase inhibitor protein), and the transmembrane protein TETRASPANIN 8 (TET8). There were five lysosome-stored cysteine protease genes: Cathepsin B-like cysteine proteinase 5 (Cpr-5), senescence-associated gene (SAG12-2), thiol protease SEN102 (SEN102), cathepsin B-like cysteine proteinase (AT4G01610) and cathepsin F-like cysteine proteinase (RD21). The three microtubule-associated proteins (MAPs) were Katanin p80 (KTN80), Ca2+-binding protein 1(CP1), and Ca2+ binding protein with one EF-Hand Motif (KIC). Additionally, one gene of α-L-fucosidase activity is FUC1.From this enrichment analysis of DEGs, we identified nineteen upregulated genes related to the respiratory chain, lysosome-stored cysteine proteases, microtubule-associated protein and α-L-fructosidase activity b. There F1F0, TET8, CYTC-2, COX, SAG12-2, RD21, Cpr-5, CP1, KIC, KTN80, AOX2, AOMI and AOX1A) were used to validate gene expression patterns in a mixed sample of OE-6 and OE-8 SAM by qRT-PCR. Compared to relative expression levels in the WT mixed samples, all measured genes were significantly upregulated in transgenic samples  family, and AP1 (APETALA1), AP3 (APETALA3), SEP1 (SEPALLATA1), SEP2 (SEPALLATA2), MADS6, TDR4 (a AGL8 homolog), and CMB1  from 10 significantly downregulated genes were selected for expression level quantification in a mixed sample of OE-6 and OE-8 SAM with qRT-PCR. The results show that those eight genes were significantly downregulated in the OE plants compared with the WT . A total of 2677 genes  were identified as core differentially expressed genes (DEGs) in  tobacco . Among t tobacco . AnalysiGhKTI12 transgenic tobacco plants, nineteen genes were significantly upregulated and related to plant development which belong to respiratory chain, microtubule organizing center part, lysosome and α-L-fucosidase activity  [KIC in Arabidopsis could regulate leaf trichome morphogenesis through facilitating the microtube-stimulated KCBP protein [2+ levels could promote primary root growth by mediating auxin signaling in KIC transgenic plants [GhKTI12 transgenic tobacco lines maybe caused by the upregulation of microtubule organizing center part related genes  ,48,49. O protein . Otherwic plants ,52,53. Sed genes . MeanwhiC1A involved in multiple physiological processes such as plant growth seed germination, another development, programed cell death (PCD), abiotic stress, and immunity [GhKTI12 transgenic tobacco lines, five cathepsin proteins genes were upregulated, such as Cpr-5, SAG12-2, SEN102, AT4G01610 and RD21A. These genes could regulate leaf senescence and plant growth. For example, the senescence-specific cysteine protease gene SAG12 negatively regulates leaf cell death. The downregulation of OsSAG12-1 resulted in stress-induced cell death in transgenic rice RNAi lines [AtSAG12 promoter delays senescence in Arabidopsis, lettuce, and wheat [NtSAG12 protein in GhKTI12 transgenic tobacco plants. A prolonged life stage with increased chlorophyll and a delay in leaf senescence could increase the photosynthetic rate and promote plant development in tobacco overexpressing GhKTI12.In the lysosome, the cathepsins are cysteine proteases of papain-like immunity . In our Ai lines . Severalnd wheat ,57,58. Ind wheat . TherefoGhKTI12 in tobacco could positively regulate cellular component-related genes to promote cell proliferation and plant development. We propose that these upregulated genes promote plants’ development of larger leaves, increased plant height, and larger root systems compared to the WT.Combined gene expression profiles with morphology data demonstrated that overexpression of GhKTI12 transgenic tobacco plants, 10 downregulated genes were identified by RNA-seq analysis in OE-6 and OE-8 lines. Eight of the downregulated genes were MADS-box transcription factor family proteins involved in flowering time regulation, such as AP1, AP3, AGL6, AGL8, AGL14, SEP et al. The gene expression pattern was validated by qRT-PCR (OE-5, OE-6 and OE-8 plants displayed late flowering, delayed by 28–30 days compared with the WT d. This i qRT-PCR .AP1 and AP3 can act either alone or in combination to determine the activation of the floral organ-specific gene in specific regions of the developing flower [AP1 plays a crucial role in the transition from vegetative to reproductive phase and floral development [AP3 determines the identity of petals and stamens [The g flower ,61. AP1 elopment , and act stamens . Overexp stamens , apple [ stamens  and soyb stamens .AGL6, AGL8 and AGL14 are pivotal in promoting floral meristem development by limiting stem cell proliferation [AP1 and SEP in Arabidopsis [AGL6 in Arabidopsis and overexpression of the SEP-like gene TaMADS1 in wheat [xal2/AGL14 causes early flowering, whereas a knockout mutant shows late flowering in Arabidopsis [Furthermore, feration ,68. AGL6bidopsis . Accordiin wheat  exhibitebidopsis .GhKTI12 transgenic plants were caused by the downregulation of MADS-box genes such as AP1, AGL6, AGL14, and SEP genes. Based on the molecular mechanism of morphology and later flowering phenotype in transgenic plants, we hypothesize that the overexpression of GhKTI12 in tobacco could create a strong basis for plant reproduction and yield by increasing plant size and delaying the flowering time with a longer vegetative stage.These results proved that a delayed flowering time and longer vegetative stage in GhKTI12 OE lines. The average dry weight of leaves and stems increased by ~34% in OE-5, OE-6 and OE-8 (GhKTI12 promotes whole plant development to increase biomass. Transgenic plants additionally produced more flowers and capsules; the average capsule number per ten plants in OE-5, OE-6 and OE-8 was 114, 128 and 142, contributing to an increase of 35%, 40% and 44% net seed yield, respectively (Biomass and seed yield were significantly increased in and OE-8 , and theand OE-8 . These rectively d,e. Howeectively .GhKTI12 could increase starch accumulation via sucrose breakdown in younger leaves of the top canopy before floral transition, which may contribute to an improved photosynthetic rate, and in turn to an increased seed yield.As the main product of photosynthesis, sucrose is transported from the source organ to the sink organ in plants and is related to biomass and yield . We founRD21 physically interacts with water-soluble chlorophyll proteins, which are involved in embryo development and seed formation via controlling cell death during the flower development progress [GhKTI12 overexpressing tobacco may also be caused by the upregulation of RD21. Taken together, these results suggest that the overexpression of GhKTI12 could improve seed yield and biomass production by regulating gene expression involved in various biological processes including cell division, leaf senescence, programed cell death, photosynthesis, and embryo development.Furthermore, the cellular component-related protein progress . TherefoGhKTI12 in tobacco led to significant variation in agriculturally important phenotypes, increasing cell proliferation and organ growth, delaying flowering time, and increasing plant height, seed yield, and biomass production. The OE-5, OE-6 and OE-8 lines with significantly improved phenotypes illustrate that GhKTI12 acts as a super-gene to positively regulate plant development and yield. Thus, GhKTI12 has broad prospects for the genetic improvement of crops in the future.In conclusion, the overexpression of


PPARD, ACOX1, ADIPOQ, CPT1A, FABP5, ASBG2, LPL, PLIN2 and related miRNAs, including mir-200b-3p, mir-130b-3p, mir-215-5p, mir-122-5p, mir-223 and mir-125b-5p, and played an important regulatory role in biological processes such as lipid metabolism, adipocyte proliferation and differentiation. This study described the dynamic expression profile of lncRNAs in the abdominal fat of Gushi chickens for the first time and constructed the DE-lncRNA interaction regulatory network. The results expand the number of known lncRNAs in chicken abdominal fat and provide valuable resources for further elucidating the posttranscriptional regulatory mechanism of chicken abdominal fat development or deposition.Chickens are one of the most important sources of meat worldwide, and the growth status of abdominal fat is closely related to production efficiency. Long noncoding RNAs (lncRNAs) play an important role in lipid metabolism and deposition regulation. However, research on the expression profile of lncRNAs related to the development of abdominal fat in chickens after hatching and their interaction regulatory networks is still lacking. To characterize the lncRNA expression profile during the development of chicken abdominal fat, abdominal adipose tissues from 6-, 14-, 22-, and 30-week-old Chinese Gushi chickens were herein used to construct 12 cDNA libraries, and a total of 3,827 new lncRNAs and 5,466 previously annotated lncRNAs were revealed. At the same time, based on the comparative analysis of five combinations, 276 differentially expressed lncRNAs (DE-lncRNAs) were screened. Functional enrichment analysis showed that the predicted target genes of these DE-lncRNAs were significantly enriched in pathways related to the posttranscriptional regulation of gene expression, negative regulation of cell proliferation, cell adhesion and other biological processes, glycosphingolipid biosynthesis, PPAR signaling, fatty acid degradation, fatty acid synthesis and others. In addition, association analysis of the lncRNA transcriptome profile was performed, and DE-lncRNA-related lncRNA-mRNA, lncRNA-miRNA and lncRNA-miRNA-mRNA interaction regulatory networks were constructed. The results showed that DE-lncRNA formed a complex network with PPAR pathway components, including  Abdominal fat is the main type of fat tissue in chickens. In the past few decades, the growth rates and body weights of chickens have increased, resulting in the excessive deposition of body fat, especially abdominal fat, thereby reducing feed utilization and the rates of egg production, fertilization and hatching among other negative effects . TherefoC/EBPa, PPARγ, AdipoQ, and FABP4, thereby promoting the differentiation of preadipocytes into mature adipocytes  approved by the Animal Care and Use Committee of Henan Agricultural University, China.The Gushi chickens used as the experimental animals in this study were obtained from the Animal Center of Henan Agricultural University. A total of 200 one-day-old female Gushi chickens were raised in cages in the same environment under standard conditions used for the pure breeding conservation of Gushi chickens. In this study, three healthy chickens were randomly selected at 6, 14, 22, and 30 weeks of age, and twelve chickens were thus used in this study. The feeding method is the same as our previous research . The chi® Ultra™ Directional RNA Library Prep Kit for Illumina® . The products were then purified using a TruSeq RNASample Prep Kit v2 , and library quality was assessed on an Agilent Bioanalyzer 2100 system . Finally, the libraries were sequenced using an Illumina HiSeq 2500 platform, and paired-end reads were generated.Total RNA was extracted from the abdominal adipose tissues of Gushi chickens aged 6, 14, 22, and 30 weeks. The quality of RNA was evaluated by Microspectrophotometer and Agarose gel electrophoresis, and then used to construct cDNA library. An initial total amount of 3 μg of RNA was used for the construction of each library. First, ribosomal RNA was removed with the Epicentre Ribo-zero™ rRNA Removal Kit , and sequencing libraries were then generated using rRNA-depleted RNA with the NEBNextftp://ftp.ensembl.org/pub/release-104/fasta/gallus_gallus/dna/). An index of the reference genome was constructed using Hast2, and paired-end clean reads were aligned to the reference genome using Hast2  value < −1.5 and a q-value < 0.05 indicated downregulated DE-lncRNAs  analysis was performed using DESeq2 software V1.22.2. The thresholds of a log2 fold change (FC) value > 1.5 and a -lncRNAs .p-value  < 0.05 were considered to be significantly enriched  > 0.7 was used to predict the regulation of trans target genes by lncRNAs. Subsequently, gene ontology (GO) and KEGG pathway analyses of the predicted target genes of each group of DE-lncRNAs were performed using the Clusterprofiler package, and GO terms and KEGG pathways with a corrected enriched .The transcriptome (PRJNA551368) and small RNA (PRJNA528858) library were constructed using the same RNA samples collected from the abdominal adipose tissues of Gushi chickens at the four developmental stages in our previous research. The association analysis of DE-lncRNAs was performed using these data . miRanda® 96 qRT-PCR system  with ChamQ Universal SYBR qPCR Master Mix . β-actin were used as internal reference genes for mRNA. The relative expression levels were analyzed with the 2−ΔΔCt method. Experiments were repeated at least three times, and all data are presented as the mean ± standard error of the mean. Data were analyzed and plotted with Prism software . The qRT-PCR primer sequences are listed in To verify the accuracy of the data obtained by high-throughput sequencing, four lncRNAs were randomly selected, and the results were confirmed by qRT-PCR for nine randomly selected lncRNAs. The RNAs were reverse transcribed into cDNA with HiScript II Q RT SuperMix for qRT-PCR (+gDNA wiper) . qRT-PCR was performed on a LightCyclerp < 0.05 was taken as the significant difference.The data were analyzed by single factor analysis of variance (ANOVA) with SPSS26.0. The difference significance test was carried out by Duncan multiple comparison. A total of 12 cDNA libraries were constructed using abdominal adipose tissues from Gushi chickens at four developmental stages. After sequencing and strict quality evaluation, each library contained 13.53–16.88 GB of clean bases, with the CG content ranging from 46.20 to 51.56%. Comparative analysis showed that 91.97–95.55% of the reads (83.08–90.16% of the number of species) were located in the reference genome. Details regarding the sequencing and quality evaluation of each library are presented in StringTie merge software was used to merge the transcripts sequenced from each library, and the merged transcripts were then screened and annotated. Finally, a total of 5,466 known lncRNAs and 3,827 new lncRNAs were identified from the 12 libraries . Among tq-value < 0.05) at four different developmental stages, namely, 6 weeks (W6), 14 weeks (W14), 22 weeks (W22), and 30 weeks (W30). Among the five different comparison groups, there were 62, 79, 140, 58, and 62 DE-lncRNAs in the W14 vs. W6, W22 vs. W6, W30 vs. W6, W30 vs. W14, and W30 vs. W22 comparison groups, respectively. However, DE-lncRNAs were not identified in the W22 vs. W14 comparison group. Venn diagram analysis showed that no DE-lncRNAs were common among the five comparison groups  were identified, and the target relationships between the DE-lncRNAs and DE-miRNAs were predicted. In total, 4,058 lncRNA-miRNA interaction pairs  were obsASBG2, LPL, and FABP5 are involved in fatty acid transport. They are related to three miRNAs , the latter two of which form 51 ceRNAs and 36 lncRNAs. In addition, PLIN2 was related to adipocyte differentiation and forms 11 ceRNAs with gga-miRNA-125b-5p and 11 lncRNAs. In addition, PPARD, gga-miR-130b-3p and 16 lncRNAs also formed 16 ceRNAs.Based on the whole-transcriptome data association analysis of abdominal adipose tissue samples from Gushi chickens at the four developmental stages, 1,740 differentially expressed lncRNA-miRNA-mRNA pairs  were scrPPARγ2 or PPARγ and inhibit the differentiation of 3T3-L1 adipocytes  of Gushi chickens at 6, 14, 22, and 30 weeks of age were 0.63 ± 0.42, 1.95 ± 1.08, 3.25 ± 1.26, and 2.96 ± 0.35, respectively. At 30 weeks, Gushi chicken had reached body maturity and was peak egg production. Due to the large energy consumption of egg production and less energy consumption for abdominal fat deposition, therefore, 14–22 weeks of age is an important stage for abdominal fat deposition in Gushi chickens. Therefore, the dynamic lncRNA expression profile revealed in this study truly reflects the molecular characteristics of Gushi chicken abdominal fat development.ACSBG2, ACOX1, PPARD, ADIPOQ, CPT1A, ACSL4, PLIN2, LPL, FABP3, FABP5 and many other key genes related to lipid metabolism or fat deposition have been confirmed to play a role in fat formation in chickens (PPARD can enhance the conversion of chicken immature adipocytes to mature adipocytes by stimulating fatty acid oxidation (CPT1A is a key factor that regulates fat deposition during chicken embryonic development and promotes chicken intramuscular growth by downregulating fatty acid oxidation and fat deposition (PLIN2 participates in the regulation of fat deposition and is significantly related to the abdominal fat weight and percentage of abdominal fat in chickens (FABP3 is significantly related to the intramuscular fat content in Beijing oil chickens and regulates intracellular fatty acid transport via a targeted interaction with lncRNA ENSGALG00000021686 (Studies have shown that lncRNAs can directly interact with genes related to lipid metabolism or with miRNAs to regulate lipid metabolism. Through these regulatory methods, lncRNAs can increase the precision and complexity of posttranscriptional lipid metabolism regulation. To reveal this complex regulatory relationship of lncRNAs during the development of Gushi chicken abdominal fat, we constructed interactive regulatory networks between DE-lncRNAs and their target gene mRNAs and miRNAs. Among the genes in the constructed lncRNA-mRNA network, chickens . For exaxidation , and CPTposition . PLIN2 pchickens . Finally00021686 . Among t00021686 ]. The abPPARD, PLIN2, FABP5, ACSBG2, LPL, miR-200b-3p, miR-130b-3p, miR-215-5p, miR-122-5p, miR-223, and miR-125b-5p, most of which have been proven to be related to fat formation. Among them, FABP5 is a member of the FABP gene family and participates in the absorption, metabolism, and transport of long-chain fatty acids (PPARG (The regulatory mechanism of lncRNA is very complex. It has been found that lncRNA located in the cytoplasm can be used as a molecular sponge of miRNA to adsorb miRNA to remove the inhibition of miRNA on its target genes. . Togethety acids , miR-130s (PPARG , and miRs (PPARG . These rIn summary, we revealed the dynamic expression profile of lncRNAs related to abdominal fat development in Gushi chickens and identified 9,293 lncRNAs and 276 DE-lncRNAs. At the same time, a DE-lncRNA-related interaction regulation network was constructed, revealing that lncRNAs mainly affect lipid metabolism or deposition, adipocyte proliferation and differentiation and other biological processes through a complex interaction network, thereby regulating the development of chicken abdominal adipose tissue. These results provide a valuable resource for better understanding the posttranscriptional regulatory mechanism underlying chicken abdominal fat development and a basis and clues for additional research on the functional mechanism by which lncRNAs regulate chicken abdominal fat deposition.


JMJD6 is a member of the Jumonji (JMJC) domain family of histone demethylases that contributes to catalyzing the demethylation of H3R2me2 and/or H4R3me2 and regulating the expression of specific genes. JMJD6-mediated demethylation modifications are involved in the regulation of transcription, chromatin structure, epigenetics, and genome integrity. The abnormal expression of JMJD6 is associated with the occurrence and development of a variety of tumors, including breast carcinoma, lung carcinoma, colon carcinoma, glioma, prostate carcinoma, melanoma, liver carcinoma, etc. Besides, JMJD6 regulates the innate immune response and affects many biological functions, as well as may play key roles in the regulation of immune response in tumors. Given the importance of epigenetic function in tumors, targeting JMJD6 gene by modulating the role of immune components in tumorigenesis and its development will contribute to the development of a promising strategy for cancer therapy. In this article, we introduce the structure and biological activities of JMJD6, followed by summarizing its roles in tumorigenesis and tumor development. Importantly, we highlight the potential functions of JMJD6 in the regulation of tumor immune response, as well as the development of JMJD6 targeted small-molecule inhibitors for cancer therapy. It also forms trimers, tetramers, and pentamers based on the JMJC catalytic domain. Human JMJD6 contains one conserved JMJC domain, five nuclear-localization sites (NLS), one DNA binding domain (AT-hook like motif), one predicted SUMOylation site, and one polyserine domain (141-Arg305) is the catalytic region of JMJD6 and forms double stranded -β helix (DSBH) fold of Fe (II)- and 2-OG-dependent dioxygenases  may facilitate the entry of functional proteins import into the nucleus  may link to the protein stability and protein-protein interactions  binds to Fe (II) and Thr184, Lys204, and Asn277 residues in the central region.JMJD6 is a 47-55kDa molecular weight member of the JMJC domain family. It is made up of 403 amino acids, and its genes are located on the 17qter chromosome . JMJD6 cygenases . Five nu nucleus . AT-Hooke domain 1-Arg305 d/or RNA . Althougof JMJD6 . The preof JMJD6 . As showmono-methylation and subsequent hydroxylation of arginine methyl groups on H3R2 and H4R3; 2) Subsequently, formaldehyde is produced through the formylation reaction, which facilitates the separation of methyl groups  database. Then R software (Version 3.6.4) was used to calculate the difference in expression between human tumor tissues and the adjacent normal tissues. Significantly upregulated levels of JMJD6 gene were observed in 17 types of tumor samples, including glioblastoma multiforme (GBM), uterine corpus endometrial carcinoma (UCEC), breast invasive carcinoma (BRCA), lung adenocarcinoma (LUAD), esophageal carcinoma (ESCA), stomach and esophageal carcinoma (STES), kidney renal papillary cell carcinoma (KIRP), pan-kidney cohort  (KIPAN), colon adenocarcinoma (COAD), colon adenocarcinoma/rectum adenocarcinoma esophageal carcinoma (COADREAD), stomach adenocarcinoma (STAD), head and neck squamous cell carcinoma (HNSC), kidney renal clear cell carcinoma (KIRC), lung squamous cell carcinoma (LUSC), liver hepatocellular carcinoma (LIHC), rectum adenocarcinoma (READ), cholangiocarcinoma (CHOL)  in triple negative breast cancer (TNBC) cells, then autophagy regulated by JMJD6- H2A.XY39ph axis promoted cell growth -V7. A phase II clinical trial showed that DNA-repair defects are more common in AR-V7-positive prostate cancer, which enables AR-V7-positive tumors more sensitive to immune checkpoint blockade , 48. Intvia the regulation of JMJD6-mediated H2A.XY39ph  promoter and inhibited its expression in the absence of PRMT1 , whereasvia downregulating H4K16ac, and ultimately modulates the DNA damage response in radiation-induced osteosarcoma cells , head and neck squamous cell carcinoma (HNSCC), esophageal squamous cell carcinoma (ESCC), osteosarcoma, acute myeloid leukemia (AML), and renal cell carcinoma (RCC). JMJD6-IL4 axis may prompt changes in clinical phenotypes of OSCC cancer stem cells, JMJD6 accelerates self-renewal activity, migration/invasion, and drug resistance . High lema cells . JMJD6 pma cells . Besidesma cells . In addima cells . In RCC, H3K27ac .Increasing evidence suggests that epigenetic events and epigenetic genes are activated or suppressed in a variety of immune cells or immune responses, leading to the occurrence of different types of diseases , 65. DatArginine methylation is a common post-translational modification that regulates a variety of functions, including cell cycle regulation of RNA processing and DNA replication. Meanwhile, arginine methylation affects gene transcription and splicing, and signal transduction, including some immune regulatory pathways . Tumor nhttps://xenabrowser.net/)), and extracted the expression data of JMJD6 and 150 immunoregulatory genes, 60 immune checkpoint-related genes as well as 22 tumor-associated immune cells in pan-cancers. Bioinformatics analyses aimed at describing the immune role of JMJD6 are pivotal in determining different types of cancers that might benefit from the anti-JMJD6 immunotherapy. As we all know, TME is a very complex environmental system related to tumor cell survival, including immune cells, fibroblasts, vascular endothelial cells, extracellular matrix and various signaling molecules. It promotes the expression of some oncogenes and inhibits immune cells from performing their immune functions. Bioinformatics analysis results uncovered that JMJD6 was negatively associated with a series of immunoregulatory genes  in LUSC, HNSC, UCS, MESO, THYM, DLBC, UCEC, LIHC, and SARC  and 2-OG-dependent manner , 71, 72.As an epigenetic factor, the role of JMJD6 in tumors has been proved to be related to the regulation of some crucial target molecules, including p53, Wnt, MAPK, Ras, ERK, TGF-β, CARM1, c-Myc, CDK4, ZFPM2-AS1, AR-V7, estrogen signal, etc. These targets modulate immune responses in TME on different levels. For example, p53 is closely related to the function and immune response of the body’s immune system . On the JMJD6 is a dioxygenase dependent on 2-OG and Fe (II). Its central JMJC domain has specific catalytic sites that target 2-OG/Fe (II) and regulate transcription and splicing by catalyzing the demethylation of histone/non-histone arginine or lysine hydroxylation , 20. The50 value of 0.78 μM against JMJD6, was firstly reported by Zheng et al. , which has good selectivity (>100 fold)  . Howevervia increasing sVEGFR-1 production of tumor-associated or GM-CSF treated macrophages  may be potential JMJD6 inhibitors . AdditioFurthermore, it also was reported that 2,4 PDCA, 2,4 BPDCA, Daminozide, the structurally related succinyl hydroxamic acid derivative 3, IOX1, and 8-hydroxyquinoline derivative 2, maybe potential inhibitors of JMJD6 . Among tin vitro and tumor inhibition in mice with neuroblastoma. These findings suggest that THZ1 and panobinostat are nonspecific inhibitors of JMJD6 (Co-inhibition of THZ1 (the CDK7/super-enhancer inhibitor) and panobinostat (the histone deacetylase inhibitor) could synergistically suppress JMJD6, resulting in increased cell apoptosis of JMJD6 .In summary, JMJD6 is involved in transcriptional chromatin structural epigenetic and genomic integrity regulation through selective demethylation. A large number of studies have shown that JMJD6 expression is increased in many tumors and contributes to the development and progression of tumors. In this article, we have discussed the biological role of JMJD6 and its important role in tumorigenesis, as well as immune response Figure 6KW drafted the manuscript and drew the original artwork. KW, CY, ZX, ZM, and HW revised the manuscript. HL, XL, and MZ edited the manuscript and contributed to the systematic evaluation of articles and literature. All authors made substantial contributions to the manuscript and approved the submitted version.This work was supported by Luzhou Science and Technology Bureau (2021-JYJ-55), the Research Fund of Southwest Medical University , and the Start-Up Research Funding of Southwest Medical University (41/00040179).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Eosinophils are important effector cells involved in allergic inflammation. When stimulated, eosinophils release a variety of mediators initiating, propagating, and maintaining local inflammation. Both, the activity and concentration of secreted and cytosolic phospholipases (PLAs) are increased in allergic inflammation, promoting the cleavage of phospholipids and thus the production of reactive lipid mediators. Eosinophils express high levels of secreted phospholipase A2 compared to other leukocytes, indicating their direct involvement in the production of lipid mediators during allergic inflammation. On the other side, eosinophils have also been recognized as crucial mediators with regulatory and homeostatic roles in local immunity and repair. Thus, targeting the complex network of lipid mediators offer a unique opportunity to target the over-activation and ‘pro-inflammatory’ phenotype of eosinophils without compromising the survival and functions of tissue-resident and homeostatic eosinophils. Here we provide a comprehensive overview of the critical role of phospholipase-derived lipid mediators in modulating eosinophil activity in health and disease. We focus on lysophospholipids, polyunsaturated fatty acids, and eicosanoids with exciting new perspectives for future drug development. Eosinophils are multifunctional leukocytes involved in the host defense against helminth infections, tissue homeostasis, and tissue repair ,2,3. EosRecent breakthroughs in eosinophil targeting have been made in the field of biologics and have been discussed in detail elsewhere . The appRecently, however, it has been recognized that eosinophils are crucial for local immunity and repair (LIAR hypothesis), by promoting immunoregulation and tissue homeostasis ,2,10. An2 in comparison to other leukocytes, indicating their involvement and contribution to lipid mediator production during allergic inflammation .,42.2–sPLof IL-33 . Even thoduction .2 have long been recognized as vital for eicosanoid production, since inherited cPLA2 deficiency leads to widespread decreases in eicosanoid levels in humans [2 -IV in eosinophil over-activation have been demonstrated in preclinical models, and cPLA2 inhibition prevented both eosinophilic infiltration and subsequent airway hyperresponsiveness after antigen challenge [2-IV is involved in the augmented release of leukotriene C4 from isolated eosinophils [Cytosolic phospholipases An humans . Furthern humans . The invhallenge . Moreoveinophils .2 inhibitors for treatment of asthma and allergic diseases has been underway for the past two decades, with many orally-bioavailable compounds entering human clinical trials [2 inhibitor ZPL-521 has now completed a phase IIb clinical trial for treatment of atopic dermatitis [2 is their high lipophilicity, which leads to unfavorable absorption, distribution, metabolism, and excretion [2-IV oxo-ester inhibitors with a favorable lipophilic profile was developed and characterized by Kokotou et al. [Drug development of specific cPLAl trials ,85,86 , thus producing lyso-PAF  . A minorPAF was shown to act on a specific G protein-coupled receptor, inducing eosinophil activation, calcium flux, and prostanoid secretion ,92. FurtThe most common lysophospholipid produced by the action of phospholipases is lysophosphatidylcholine (LPC), which can reach plasma concentrations of up to 150 µM already under physiological conditions . LPC is Studies investigating the effects of LPC on immune cells are contradictory . SpecifiIn fact, Kwatia et al. reported that a combined action of both secretory phospholipases and eosinophil-derived lysophospholipases is needed to induce a loss in surfactant activity as observed in asthma . Moreove2+ flux and kinase phosphorylation [2, which may be another anti-inflammatory mechanism [Saturated LPC species added to human eosinophils in the presence of the physiological carrier, serum albumin, strongly suppressed upregulation of CD11b and degranulation . The antrylation . Short-trylation . Interesechanism .Consistent with the anti-inflammatory effects observed with LPC , the sta10 receptor [10 is highly expressed during late stage eosinophil differentiation and in eosinophils isolated from severe asthmatics, it offers the possibility of targeting eosinophil degranulation without affecting their survival [Recently, another lysophospholipid has been investigated in the context of eosinophil activation. Hwang et al. demonstrated that lyosphosphatidylserine is capable of inducing eosinophil degranulation via its action on P2Yreceptor . A subsereceptor . Becausesurvival ,117.The most recognized polyunsaturated fatty acid (PUFA) produced by the action of phospholipases is ω-6 acid arachidonic acid  . ArachidOther important PUFAs involved in inflammation and allergy are the ω-3 acids eicosapentaenoic acid  and docosahexaenoic acid  . The supEicosanoids are a diverse group of short-lived bioactive mediators mainly metabolized from arachidonic acid, which control a wide range of physiological processes such as vasoconstriction and vasodilatation, gastric motility, and bronchoconstriction. Cells of the innate immune system such as neutrophils, macrophages, mast cells, and eosinophils are capable of producing large amounts of eicosanoids, which may potentiate or attenuate local inflammatory processes. Moreover, eicosanoids were reported to play a vital role in a variety of diseases ranging from allergy to autoimmunity and cancer. Eicosanoids predominantly bind to and activate G protein-coupled receptors (GPCRs) and therefore represent a readily targetable group of lipid mediators ,155,156.SPMs consist of E-series resolvins, D-series resolvins, protectins, maresins, as well as AA-derived lipoxins. This group of anti-inflammatory mediators inhibits polymorphonuclear leukocyte infiltration and promotes resolution of inflammation via macrophage clearance ,158,159.1 and inhibiting IL-17 and IL-23 cytokine release. This results in decreased immune cell infiltration into the airways and improved lung function parameters [4) were found to be decreased in severe asthma and associated with aspirin sensitivity in aspirin-exacerbated respiratory disease [4 receptor are decreased in severe asthma [4 [Eosinophils are a major source of resolvin E3 produced via the 12/15-LOX pathway, whereas neutrophils produce mainly resolvin E1 and resolvin E2 via the 5-LOX pathway . Eosinoprameters . Followirameters . Interesrameters ,144,159.rameters ,144,159. disease ,162,163.e asthma . Previousthma [4 .4 analogs have demonstrated safety and clinical efficacy as a topical solution for infant eczema (15(R/S)-methyl-lipoxin A(4)) [Since then, clinical studies with LXAin A(4))  and for in A(4)) .4 (LTA4). Finally, LTA4 can be further metabolized via three different pathways into cysteinyl leukotrienes , LTB4 and into anti-inflammatory lipoxins (LXA4 and LXB4) [Leukotrienes are produced via enzymatic lipid peroxidation from arachidonic acid by the action of lipoxygenases (LOX). First, diverse LOX  species convert arachidonic acid into hydroperoxyeicosatetraenoic acids (HpETEs) and HETEs. 5-LO next converts the intermediate HpETE into a transient epoxide intermediate leukotriene And LXB4) ,165.1, CysLT2), as well as GPR99, GPR17 and P2Y12 [1, CysLT2, GPR99 and P2Y12 [1 on eosinophils was found to increase their transendothelial migration, superoxide generation, and degranulation [Cysteinyl leukotrienes have been reported to bind to 5 different receptors with varying affinities–cysteinyl leukotriene receptors 1 and 2  is generated, which serves as a substrate for various synthases that form PGD2, PGE2, PGI2, PGF2α, and thromboxane [Prostaglandins are bioactive mediators formed from arachidonic acid via the cyclooxygenase pathway. First, an intermediate prostaglandin Homboxane . Since tomboxane , this se2 is metabolized from PGH2 via the action of hematopoietic PGD synthase (hPGDS) [2 is reported to bind to two receptors DP1 and DP2  [2 following stimulation [2 and express higher levels of hPGDS [In the peripheral tissues, PGD (hPGDS) ,196. PGDs–CRTH2) . Both, ts–CRTH2) ,198,199.mulation . Furtherof hPGDS . Similarof hPGDS . Finallyof hPGDS .2 on its receptors DP1 and DP2 on eosinophils have been reviewed in detail by Peinhaupt et al. [2 or selective agonists to DP2 promotes Ca2+ flux, CD11b upregulation, migration and degranulation of human eosinophils [2 was found to prime eosinophils for other chemoattractants [4 secretion from eosinophils [The multiple pro-inflammatory actions of PGDt et al. . Brieflyinophils ,119. Morractants ,121. Actractants ,122. Morinophils ,204.2 pathway in allergic inflammation, mainly DP2 antagonists have been used, while only a few reports studied the effects of hPGDS inhibition. In a recent preclinical study of allergic rhinitis, the hPGDS inhibitor TAS-205 in combination with montelukast showed a significant additive inhibitory effect on eosinophil infiltration and nasal obstruction in the late phase, when compared to treatment with either active agent alone [For targeting the PGDnt alone . Moreovent alone . CurrentIn contrast, DP receptors have been studied in detail in multiple clinical studies with mixed results ,205,206.2 is produced from PGH2 by the action of prostaglandin E synthases [2 (together with LXA4 and PGI2) is hypothesized to be an arachidonic acid-derived pro-resolving mediator [2 is considered detrimental in the tumor microenvironment [2 were found to be increased in asthma and non-asthmatic eosinophilic bronchitis, while on the other hand, Aggarwal et al. reported an inverse relationship between PGE2 levels and eosinophil sputum counts [2 is reported to activate four receptors . Of those, eosinophils were reported to express EP2 and EP4 receptors [PGEynthases . While Pmediator , it alsomediator . Moreoveironment ,210,211.m counts ,206,212.eceptors ,213,214.eceptors .2 inhibits their effector responses such as chemotaxis and degranulation as well as CD11b upregulation [2 was also found to promote survival of eosinophils [2-EP4 axis is important for the interactions of eosinophils with endothelial cells [2 prior to allergen challenge significantly improved lung function parameters and attenuated airway inflammation and reduced eosinophil counts in sputum [2 suppresses eosinophil activation, PGE2 can activate other immune cell types and therefore may promote the development of diseases such as atherosclerosis, rheumatoid arthritis, cancer, and influenza virus infections [2-induced impaired immune response, male sex, and the risk of cardiovascular complications in COVID-19 patients [Upon binding to EP2 and EP4 on eosinophils, PGEgulation ,124. Surinophils ,126. By al cells . Finallyal cells , and in al cells ,217. In n sputum . Althougfections ,221,222.patients . Thus, a2) is synthesized from PGH2 via the action of prostaglandin I synthase [2–iloprost is currently indicated for treatment of pulmonary arterial hypertension [2 on the vasculature have been thoroughly studied in the past, its immunomodulatory functions have only recently been recognized and are currently being studied in more detail [Prostacyclin , preventing house dust mite-induced allergic inflammation [Overall, IP-receptor deficient mice exhibited exacerbated allergic inflammatory responses and airway remodeling, while inhalation of iloprost improved features of asthma in a preclinical model ,226,227.ammation .5-oxo-ETE is a lipid mediator produced from AA via oxidation of the 5-LO intermediate 5-HETE by the 5-hydroxyeicosanoid dehydrogenase (5-HEDH). 5-HETE has been shown to induce neutrophil and eosinophil migration and activation in vitro and in vivo and increased 5-HETE release has been observed in alveolar macrophages from asthmatic patients . Compare5-HETE and 5-oxo-ETE were so far reported to bind to and activate the GPCR OXER1  ,233. ExpIn contrast to research in the field of leukotrienes and prostaglandins, very few studies have been performed on 5-oxo-ETE and its receptor. This is mainly due to the absence of 5-oxo-ETE receptor and its orthologs in rodents limiting preclinical research. However, recently novel selective 5-oxo-receptor antagonists were characterized and tested ,239,240.2) at the phosphodiester bond between the glycerol backbone and the phosphate group. PLC produces diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3) in response to cell activation signals [3 and DAG are crucial molecules regulating signal transduction during cell activation and play a role in development of various diseases [2+ flux, resulting from an increase in IP3 concentration [Phospholipase C hydrolyzes phosphatidylinositol 4,5-bisphosphate  [In contrast to soluble IPl (2-AG) . Eosinopl (2-AG) .1, CB2). Moreover, 2-AG may be converted to arachidonic acid via the monoacylglycerol lipase (MGL) pathway leading to eicosanoid production [1 and CB2 receptors as well as MGL [2 receptor [2 receptor is increased in eosinophils from symptomatic allergic donors and upregulated following ovalbumin challenge in mice, while CB1 expression was found to be upregulated in tonsils from allergic rhinitis patients [2-AG belongs to the endocannabinoid system consisting of endocannabinoid ligands, endocannabinoid receptors, and enzymes related to their synthesis and degradation. While many studies depict the roles of the endocannabinoid system in allergic diseases, their actual contribution remains controversial . 2-AG bioduction . Human ll as MGL ,252,253.receptor ,251,253 receptor ,255. Furpatients ,256,257.patients ,258.2 receptor is known to mediate anti-inflammatory effects in many diseases, such as atherosclerosis and stress-induced neuroinflammation [2 receptor agonist S-777469 exhibited efficacy for treatment of atopic dermatitis in preclinical studies, supposedly via blockade of endogenous 2-AG binding to CB2. Unfortunately, results from a phase Ib/IIa clinical study in patients (NCT00697710) have not yet been made available [The CBammation ,259,260.ammation ,258. Thevailable .Another possibility of targeting the 2-AG metabolism in eosinophils arises from specific blockade of its hydrolyzing enzyme MGL. A recent report by Abohalaka et al. suggested that inhibition of MGL with the specific inhibitor JZL184 ameliorates lipopolysaccharide-induced airway inflammation . SimilarPhospholipases D cleave the phospholipid molecule yielding the phosphatidic acid and the phospholipid headgroup . Consequently, phosphatidic acid can be further metabolized into lysophosphatidic acid (LPA). Another pathway resulting in LPA production involves the hydrolysis of LPC via lysophospholipase D  . Multipl1–6)—the so-called canonical LPA receptors [1 and LPA3 receptor, but lack LPA2 receptor expression [2+ flux, and CD11b upregulation [2, which reportedly suppresses airway hyperresponsiveness and immune cell infiltration in the BALF and lung tissue [Lysophosphatidic acid is reported to bind to and activate six different GPCRs  or mice lacking RAGE expression) [Additionally, LPA is reported to activate non-canonical receptors such as peroxisome proliferator-activated receptor (PPAR)-γ, vanilloid receptor 1 channel as well as the receptor for advanced glycosylation products (RAGE) ,256,257.ression) ,269. In ression) ,271,272.The role of autotaxin and LPA in immune regulation and asthma was recently reviewed by Kim et al. . In thei2·−,·OH, H2O2, hypochlorous and hypobromous acids [Exposure to allergic stimuli as well as gaseous pollutants, chemicals, viruses, and bacteria leads to the recruitment of eosinophils and neutrophils, which have a high capacity for reactive oxygen species (ROS) production. Activation of these inflammatory cells leads to the so-called ‘respiratory burst’, which is characterized by an increased production of Ous acids ,274,275.us acids ,279,280.Eosinophils are reported to release up to two times more ROS compared to neutrophils . MoreoveLipid peroxidation is generally considered as the main molecular mechanism involved in oxidative damage to cell structures. PUFAs, in particular, are prone to non-enzymatic lipid peroxidation due to the presence of the double bond near the methylene bridge, leading to autooxidation by the attack of free radicals . BioactiEosinophils contribute to oxidative stress in mild asthma and targeting oxidative stress could be considered as a treatment option during asthma exacerbations . PreclinPhospholipases produce multiple reactive cleavage products, including lysophospholipids, polyunsaturated fatty acids, and eicosanoids. The highly complex network of lipids and lipid mediators modulates a variety of eosinophil functions in health and disease. The discovery of novel subclasses of lipid mediators and their receptors opens new possibilities for drug targeting. Because lipid mediator levels and the activity of their metabolizing enzymes are dysregulated in allergic inflammation and eosinophil-associated diseases, they offer a unique opportunity to target the over-activation and ‘pro-inflammatory’ phenotype of eosinophils. Importantly, this could be achieved without compromising the survival and functions of tissue-resident and homeostatic eosinophils. Fortunately, most lipid mediators function through GPCRs, which are highly drug-responsive receptors that can be readily targeted with orally bioavailable small molecule drugs.