Adaptation to environmental variation caused by global climate change is a significant aspect of fisheries management and ecology. A reduction in ocean salinity is visible in near-shore areas, especially in the Baltic Sea, where it is affecting the Atlantic cod population. Cod is one of the most significant teleost species, with high ecological and economical value worldwide. The population of cod in the Baltic Sea has been traditionally divided into two subpopulations (western and eastern) existing in higher- and lower-salinity waters, respectively. In recent decades, both Baltic cod subpopulations have declined massively. One of the reasons for the poor condition of cod in the Baltic Sea is environmental factors, including salinity. Thus, in this study, an oligonucleotide microarray was applied to explore differences between Baltic cod subpopulations in response to salinity fluctuations. For this purpose, an exposure experiment was conducted consisting of salinity elevation and reduction, and gene expression was measured in gill tissue. We found 400 differentially expressed genes (DEGs) involved in the immune response, metabolism, programmed cell death, cytoskeleton, and extracellular matrix that showed a subpopulation-dependent pattern. These findings indicate that osmoregulation in Baltic cod is a complex process, and that western and eastern Baltic cod subpopulations respond differently to salinity changes. Gadus morhua L.).Global climate change due to human activities has an impact on many environmental parameters, including salinity. Salinity fluctuation is a significant climate change stressor affecting the abundance, diversity, distribution, growth, osmolarity, and metabolic activities of fish and other marine organisms . SeawateThe Atlantic cod is a key euryhaline fish species in the North Atlantic Ocean, with significant commercial and ecological value that will be threatened by the expected climate-related decline in salinity in the Baltic Sea ,11. Cod −1, Baltic cod is exposed to variable salinities [atp1a) and heat shock protein 70 (hsp70), aquaporins, the solute carrier gene family, and receptors of prolactin (prl) [Adult Baltic cod often undertake complex migrations to feeding and spawning grounds depending on the area . Accordilinities . For suclinities . Due to linities , sperm mlinities , hemoglolinities , osmoreglinities , and diflinities . Studiesin (prl) ,31,32. Fin (prl) ,18,33. Tin (prl) . Our prein (prl) ,34. Howein (prl) ,35. The Although many studies have investigated the response of the fish transcriptome to salinity changes using next-generation sequencing (NGS) ,37,38 anThe experimental procedures were carried out according to the EC Directive 2010/63/EU for animal experiments and were approved by the Local Ethics Committee on Animal Experimentation of Gdansk Medical University (decision no. 60/2012).Gadus morhua juvenile individuals were caught in November 2012 in Schleifjord, Kiel Bight (KIEL)—ICES SD 22, with an average size of 33.95 ± 0.32 cm, and the Gulf of Gdańsk (GDA)—SD 26, with an average size of 30.7 ± 5.27 cm, which represent the western and eastern Baltic cod subpopulations, respectively , in a natural photoperiod, and under salinity conditions similar to those of the collection sites . They were fed fresh herrings once a day and acclimated for more than 14 days until their typical behavior was restored. Next, fish from each localization were randomly divided and transferred to separate tanks termed control, salinity decrease, and salinity increase groups, and again acclimated as described above . The conSamples of gill were collected from 81 fish: KIEL (n = 48) and GDA (n = 33) and stored in RNAlater, according to the manufacturer’s instructions  . Total Rp-value < 0.05, and a false discovery rate (FDR) < 0.05. The Pearson correlation between experimental groups was analyzed using the R package. The Wilcoxon rank sum test (in R) was used to compare the means of the GDA and KIEL groups. Normalized data were analyzed using a two-factor model implemented in the limma package in R  to assess the time course of responses to salinity changes in subpopulations .Microarray hybridization was performed in the Department of Physiological Sciences of Warsaw University of Life Sciences (SGGW) with the Agilent-0048047 Genome-wide Atlantic cod microarray, second version_ACIQ2  using the Gene Expression Hybridization Kit , according to the manufacturer’s protocol. This DNA oligonucleotide 4 × 44 k microarray includes 60 mer probes for unique transcripts from the Ensembl and Unigene databases . Normalihttp://www.ensembl.org/Gadus_morhua, accessed on 1 February 2020). Genes without annotations were searched against proteins from the NCBI non-redundant (nr) database using the Basic Local Alignment Search Tool (blastx) implemented in BLAST+ (v.2.2.29) [−5, using sequences to which probes were designed [Kyoto Encyclopedia of Genes and Genomes (KEGG) Orthology-Based Annotation System (KOBAS v.3.0) for the human data  [Available gene annotations were downloaded from the Ensembl database  , with andesigned . Gene sy < 0.05) .The microarray data were deposited in the National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO) under the accession number GEO: GSE195878.p-value < 0.001; The length and weight of each fish used in this experiment were measured and compared between the GDA and KIEL groups using the Wilcoxon rank sum test. Statistical analysis revealed that the body parameters of the GDA fish were significantly lower than those of the KIEL group  in both the KIEL and GDA groups . In summary, 756 DEGs and 881 DEGs were identified in the GDA and KIEL subpopulations, respectively . Of thesA groups b.p-value < 0.05; p-value < 0.001), following which salinity decreased . Salinity increased by 10 ppt caused similar magnitude expression changes in both the GDA and KIEL groups at each time point  performed for all identified DEGs from this study showed a clear division of the GDA and KIEL groups .cd59, complement factor H (cfh), chemokines , mucins , lectins toll-like receptor 2 (tlr2), nlr family card and pyrin domain containing 3 (nlrc3 and nlrp3), and natterin (aep1). Endocrine system pathways, such as thyroid hormone signaling, PPAR signaling, and estrogen signaling, were also enriched across subpopulation-dependent genes. Between these genes, there were several Ca2+-transporting ATPases (atp2a1 and atp2a2), protein kinase c alpha (prkca), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (pik3ca), notch receptor 2 (notch2), iodothyronine deiodinase 1 (dio1), and creb-binding protein (crebbp). The EBC and WBC subpopulations also showed differential regulation of genes involved in lipid metabolism , such as ceramides , phospholipase a and acyltransferase 3 and 4 (plaat3 and plaat4), phospholipase d family member 4 (pld4), prostaglandin i2 synthase (ptgis), farnesyl diphosphate synthase (fdps), fatty acid-binding protein 1 (fabp1), acyl-coa synthetase bubblegum family member 2 (acsbg2), and two genes of the cytochrome p450 family 1 subfamily a (cyp1a1 and cyp1a2). Furthermore, genes involved in programmed cell death , including caspases , bcl2-associated x, apoptotic regulators (bax), bcl2-like 14 (bcl2l14), bcl2-interacting protein 3 (bnip3), and members of the tnf receptor superfamily , were differentially expressed between these two subpopulations. The western and eastern subpopulations also showed differential regulation of genes related to tissue remodeling, such as type X collagen (col10a1), type 13 keratin (krt13), fibronectin 1 (fn1), lysyl oxidase (lox), tenascin n (tnn), myosin heavy chain 3 and 7 (myh3 and myh7), and myosin light chain 3 and 10 (myl3 and myl10), which are involved in the regulation of the actin cytoskeleton  showed differential expression between subpopulations FC > 2; . Of thesskeleton .bax, casp8, kinesin family member 5b (kif5b), nlrp3, pik3ca, and tnfrsf10b involved in apoptosis and necroptosis, cyp1a1, cyp1a2, and acsbg2 involved in fatty acid metabolism, and dio1 and notch2 related to the thyroid hormone signaling pathway. Additionally, several immune-related genes such as aep1, ccl20, and interferon-induced very large GTPase 1 (gvinp1), while fish-egg lectin, muc2, and piscidin were regulated over time , sam and hd domain containing deoxynucleoside triphosphate triphosphohydrolase 1 (samhd1), tnfrsf14, and tnfrsf15, as well as the stress response protein (hsp70) and genes involved in glycerophospholipid biosynthesis (plaat3 and plaat4) and haloacid dehalogenase-like hydrolase domain containing 5 (hdhd5) were differentially expressed only 1 h after the change in salinity. Further, 12 h after the salinity change, genes involved in neutrophil degranulation, such as rab37, members of the ras oncogene family (rab37), actin-related protein 2 (actr2), purine nucleoside phosphorylase (pnp), and tlr2, showed differences between the GDA and KIEL groups, as well as mhc class I anigen. Moreover, genes involved in the cell cycle, including the up-regulator of cell proliferation-like (urgcp) and nedd1 gamma-tubulin ring complex targeting factor (nedd1), and in tyrosine degradation, such as dihydropteridine reductase (qdpr) and asparaginase and isoaspartyl peptidase 1 (asrgl1), were only differentially expressed after 12 h. On the other hand, aldolase fructose-bisphosphate a  (involved in glycolysis), pld4 (involved in inositol phosphate metabolism), tripartite motif containing 39 (trim39), sialic acid-binding ig-like lectin 14 (siglec14), and killer cell lectin-like receptor b1 (klrb1) (involved in the innate immune system) were only differentially expressed 72 h after the salinity change. Of the 308 subpopulation-dependent genes related to salinity change, the highest number of DEGs was observed after salinity decreased (190) and the lowest after salinity increased to 33 ppt. Of these, 74 DEGs (23.94%) were shared between the salinity groups. According to the time course, salinity increased and decreased caused a differential expression of genes involved in the immune response , thyroid metabolism (dio1 and notch2) and ion channel transport  , col10a1, and fn1, were common between salinities. On the other hand, genes related to the cell cycle, including blm recq-like helicase (blm) and baf nuclear assembly factor 1 (banf1), response to elevated platelet cytosolic Ca2+ , immune-related genes , and plaat4 (involved in glycerophospholipid biosynthesis), were only differentially expressed at 3 ppt. Salinity increased by 10 ppt (to 18 ppt and 28 ppt) caused caspase expression changes (casp2 and casp8), bcl2l14 related to the apoptosis process, some immune-related genes (ccl2 and ccl20), immunoglobulin lambda constant 1 (iglc1), and apolipoprotein L domain-containing protein 1 (apold1) (involved in lipid metabolism). The lowest number of genes that showed a difference between the GDA and KIEL groups was detected at a salinity level increased to 33 ppt. Between identified genes were those involved in the notch signaling pathway (notch2 and crebbp), ppar signaling and lipid metabolism (acsbg2 and fabp1), ferroptosis  and the keratinization process, such as keratin (krt13), cornifelin (cnfn), and s100 calcium-binding protein a16 (s100a16).Of the total of 290 DEGs with time differences, 75 (25.86%) were shared between time points, including such transcripts as ver time a. A suit; trpv5) b. AdditiSalinity is an important abiotic factor that affects cod activity, distribution, and reproduction in the Baltic Sea. Due to climate change and increased precipitation in northern latitudes, salinity in the Baltic Sea has been expected to decrease . Areas wPrevious studies on the adaptation of Atlantic cod to low salinity in the Baltic Sea showed that the mechanisms may be different in divergent subpopulations of cod, which may contribute to a strong and effective reproductive barrier ,16,34. Hcsl2-like and sml-like) showed a subpopulation-dependent pattern during salinity fluctuations throughout the course of time, while fish egg lectins (fels) were regulated during salinity elevation. These results suggest that different types of CLR are activated during salinity fluctuations in Baltic cod. In addition to pattern recognition, lectins are involved in inflammation, agglutination, phagocytic reactions, cell proliferation, protein folding, RNA splicing, and molecular trafficking of molecules [nlrp3 and nlrc3) showed a subpopulation-dependent pattern under all salinities and times. NLRs play a crucial role in the inflammatory response by forming an inflammasome in the cytosol to protect the organism from pathogens and environmental factors [Ctenopharyngodon idella) [cfh, cd59, and plat (involved in complement cascade) showed different expression patterns between the EBC and WBC subpopulations. Complement components recognize, opsonize, phagocyte pathogens and promote the inflammatory response [Oncorhynchus kisutch) gill tissue, members of this pathway were both down- and up-regulated in response to salinity stress [lysozyme g showed differential expression between the EBC and WBC subpopulations. The lysozyme is a lytic enzyme that lyzes pathogens, and is involved in opsonization, phagocytosis, and activation of the complement cascade [Takifugu fasciatus and common carp (Cyprinus carpio) revealed that salinity had an impact on lysozyme activity [lysozyme g activity was associated with complement activity in Tilapia (Oreochromis mossambicus) after transfer to saltwater [piscidin belonging to teleost-specific antimicrobial peptides (AMPs) showed a subpopulation-dependent pattern as salinity declined. Piscidin is involved in the innate immune response and exhibits antimicrobial activity against various bacteria, viruses, fungi, and parasites [piscidin can alter plasma membranes via pore formation [Muc2 was common among all salinities, I-muc showed subpopulation-dependent patterns at low salinity, while muc5ab and muc5b during salinity increased. Further, st3 beta-galactoside alpha-2,3-sialyltransferase 1 , involved in mucin type O-glycan biosynthesis, showed different expression patterns between the GDA and KIEL groups. Previous studies on Atlantic salmon  have revealed that the glycosylation process may differ within and between species [Aphanius dispar) [Anguilla japonica) after their transfer to seawater [aep1 were identified during salinity fluctuations, and most of them showed subpopulation-dependent patterns. Aep1 belongs to the pore-forming proteins (PFPs), widely distributed among teleost genomes, which efficiently kill targets due to the damage to membrane cells by forming a transmembrane pore [Danio rerio) showed that they function as innate immune molecules [Salvelinus alpinus) [The analysis of gene expression revealed many subpopulation-dependent genes involved in immune defense in WBC and EBC, thus indicating that salinity fluctuations have a complex effect on the immune system, which is in accordance with previous studies in teleosts ,40. Chanolecules . Further factors . The res idella) . In part idella) . In thisresponse . Previouresponse . In coho cascade . In accoactivity ,64. In aaltwater . Additioarasites . Previouormation , and itsormation . Togetheormation . The mucormation ,69. Othe species ,71. The  dispar) , and the dispar) . Mucus cseawater . In thisane pore ,75. Studolecules . So far,alpinus) . Togethecasp2, casp8, and casp13), proteins from the Bcl2 family , and members of the tumor necrosis factor receptor superfamily  showed a subpopulation-dependent response. Caspases are highly conserved intracellular cysteine-dependent proteases known for their critical role in mediating apoptosis and inflammatory responses [casp8 previously showed population-dependent regulation in killifish (Fundulus heteroclitus) [bax/bcl2 expression ratios can affect mitochondrial cytochrome c release [In the present study, salinity stress affected the expression of genes related to programmed cell death (PCD), including the apoptosis, necroptosis, and p53 signaling pathways. Between these genes, several caspases (esponses . Accordioclitus) . The memoclitus) . Differe release , thus in release . TNFSFs  release . The inf release  and Arab release . Accordi release ,84.col10a1, myh3, myh7, myl3, myl10, fn1, des, and lox, showed differences between the EBC and WBC subpopulations. Lox is an enzyme that plays a role in ECM remodeling by cross-linking collagen and elastin [krt13, cnfn, and s100a16) were differentially expressed in the KIEL and GDA groups in response to increased salinity. During the keratinization process, keratins accumulate inside epithelial tissue cells to form a barrier, thereby reducing water loss during dehydration, as seen in Arabian pupfish following salinity [Oncorhynchus mykiss) gill ionocytes in freshwater and seawater [In this study, genes encoding components of the cytoskeleton and extracellular matrix (ECM) involved in cell structure, motility, intercellular communication, and morphogenesis, such as  elastin . Additiosalinity . Similarsalinity . Gill epsalinity  and is aseawater .Actipenser baeri) highlighted the importance of the thyroid hormone signaling pathway in this process [dio1, prkca, pik3ca, notch2, and crebbp were differentially expressed between the EBC and WBC subpopulations. These results suggest that the impacts of salinity on thyroid hormone homeostasis and the activity of the presented genes depend on the cod subpopulation.Endocrine regulation, including thyroid hormones, is important in salinity acclimatization in euryhaline fish . Studies process , which h process . In thisacsbg2 and fabp1, involved in fatty acid metabolism, were differentially regulated during the decrease and increase. These genes were also altered in response to salinity in the delta smelt (Hypomesus transpacificus) [Cynoglossus semilaevis) [cers2, cers3, and cers4) showed a subpopulation-dependent trend at low and high salinity, depending on the transcript variant. Ceramide synthases reside at the center of sphingolipid metabolism by producing ceramides through de novo synthesis or degradative pathways [cers2 was down-regulated in Arctic charr [cyp1a1, cyp1a2, and cyp2d15 that encode cytochrome P450 (CYP) enzymes showed subpopulation-dependent patterns. CYPs are involved in the transformation and metabolism of many endogenous and exogenous compounds in fish, including the biodegradation of xenobiotics. In teleosts, cyp1a1 has been suggested as an indicator of chemical stress [Alterations in the metabolic pathways in response to salinity have been observed in various species ,91. The cificus)  and in tilaevis) . Moreovepathways . Previouic charr  and up-ric charr . In thisl stress , and itsl stress ,97. In sGene expression analysis using an oligonucleotide microarray showed the complexity of salinity adaptation in cod from the Baltic Sea, and revealed that eastern and western cod subpopulations respond differently to fluctuations in salinity. Among the subpopulation-dependent genes were those involved in the immune system, which could thus affect susceptibility to pathogens. Moreover, the EBC and WBC subpopulations differ in metabolism reprogramming, gill remodeling, and programmed cell death. This study provides the first general insight into the pathways and functional categories involved in the response of two Baltic cod subpopulations to salinity fluctuations.


The treatment provided for breast cancer depends on the expression of hormone receptors, human epidermal growth factor receptor-2 (HER2), and cancer staging. Surgical intervention, along with chemotherapy or radiation therapy, is the mainstay of treatment. Currently, precision medicine has led to personalized treatment using reliable biomarkers for the heterogeneity of breast cancer. Recent studies have shown that epigenetic modifications contribute to tumorigenesis through alterations in the expression of tumor suppressor genes. Our aim was to investigate the role of epigenetic modifications in genes involved in breast cancer. A total of 486 patients from The Cancer Genome Atlas Pan-cancer BRCA project were enrolled in our study. Hierarchical agglomerative clustering analysis further divided the 31 candidate genes into 2 clusters according to the optimal number. Kaplan–Meier plots showed worse progression-free survival (PFS) in the high-risk group of gene cluster 1 (GC1). In addition, the high-risk group showed worse PFS in GC1 with lymph node invasion, which also presented a trend of better PFS when chemotherapy was combined with radiotherapy than when chemotherapy was administered alone. In conclusion, we developed a novel panel using hierarchical clustering that high-risk groups of GC1 may be promising predictive biomarkers in the clinical treatment of patients with breast cancer. Breast cancer is the most prevalent cancer globally, with 2.26 million new cases diagnosed in 2020 . Breast Systemic treatment for non-metastatic breast cancer is based on molecular subtypes. For patients with hormone receptor-positive breast cancer, standard endocrine therapy is administered for 5–10 years, according to menopausal status . HER2-taClinically, prognostic factors can predict the population of at-risk patients, while predictive factors can identify a certain group that benefits from treatment. The strongest prognostic factors for breast cancer include tumor size, axillary lymph node involvement, histological grade, hormone receptor status, and age at diagnosis. The proportion of distant metastatic disease in breast carcinoma in situ was <1% at 10 years of age. The 5-year overall survival rates of stage I hormone receptor-positive breast cancer, HER2-positive breast cancer, and TNBC are >99%, >94%, and >85%, respectively [Currently, oncologists use molecular genomic profiling tests to establish precision medicine that can help personalize treatment, avoid over- or under-treatment, and maximize the outcomes for all subtypes of breast cancers. The following are several commercially available modalities of genomic assays that help clinicians decide on systemic therapy for early invasive breast cancer unrelated to the National Comprehensive Cancer Network, including Oncotype DX , MammaPrTo date, researchers have investigated the relationship between epigenetic alterations and tumorigenesis. Recent studies have shown that epigenetic modifications are important in human breast cancer ,20. ThisIt is thought that changes in the expression levels of these genes alter the expression of critical genes involved in cell cycle regulation, DNA damage repair, and apoptosis. These alterations may affect the sensitivity of cancer cells to chemotherapy and radiotherapy and contribute to the development of resistance. However, the mechanism behind the association of these genes with breast cancer patient outcomes and response to therapy is complex and not fully understood. In the past decade, the direction of cancer treatment is from a “one-size-fits-all” approach turn into focusing on the precision medicine based on genomic variants . TherefoA total of 486 patients were included in this study. DNMT3B/DNMT3A, HDAC2/DNMT3A, PRDM2/PRDM10, PRDM4/PRDM10, PRDM8/PRDM1, PRMT10/PRDM10, PRMT10/PRDM2, and PRMT7/PRMT1. A strong negative correlation (red) was observed between PRMT1/PRDM10, PRMT1/PRDM2, PRMT1/PRDM5, PRMT7/PRDM10, and PRMT10/PRMT1. The mRNA expression of the 31 candidate genes in the study population is shown in DNMT1, DNMT2, DNMT3A, DNMT3B, DNMT3L, HDAC1, HDAC2, HDAC3, PRDM7, PRDM9, PRDM12, PRDM13, PRDM14, PRDM15, PRMT1, PRMT2, PRMT5, PRMT6, PRMT7, and PRMT8. GC2 contained 11 genes: PRDM1, PRDM2, PRDM4, PRDM5, PRDM6, PRDM8, PRDM10, PRDM11, PRDM16, PRMT3, and PRMT10. Risk estimation subgroups according to overall patient characteristics, lymph node invasion status, and treatment were further analyzed.According to mRNA expression levels, the target genes were distributed into 2 groups, named gene clusters 1 and 2 (GC1/2). GC1 contained 20 genes, including p < 0.05), except for HDAC2, PRDM13, PRMT2, PRMT5, PRMT6, and PRDM11. In GC1, DNMT1, DNMT2, DNMT3A, DNMT3B, HDAC1, HDAC2, HDAC3, PRDM12, PRDM13, PRDM15, PRMT1, PRMT2, and PRMT5 showed positive values in the high-risk subgroup, whereas DNMT3L, PRDM7, PRDM9, PRDM14, PRMT7, and PRMT8 showed negative median values. In GC2, PRDM1, PRDM4, PRDM6, and PRMT3 had positive median values in the high-risk subgroup, whereas PRDM2, PRDM5, PRDM8, PRDM10, PRDM11, PRDM16, and PRMT10 had negative median values.The high-risk subgroups in GC1 and GC2 revealed higher mRNA expression of almost all target genes, as shown in p < 0.001). There were no significant differences in lymph node invasion, stage, or radiotherapy between the low- and high-risk groups within the two GCs. More than 90% of high-risk patients in GC1 received chemotherapy, with a significant difference (p < 0.001). Progressive disease was more frequent in the high-risk group of GC1 compared with the low-risk group .The distributions of the baseline characteristics according to hierarchical clusters are summarized in p = 0.009), with a lower PFS in the high-risk group. However, no significant differences were observed in GC2 (p = 0.543). In addition, the high-risk GC1 group with lymph node invasion showed worse PFS than the low-risk group (p = 0.003), as illustrated in p = 0.018). Interestingly, there was a trend toward better PFS in the high-risk subgroup of GC1 in the chemotherapy combined with radiotherapy setting, but the difference was not significant (p = 0.241) (p = 0.214) (The Kaplan–Meier plot illustrating the 10-year PFS is shown in = 0.241) . Additio= 0.214) .HDAC3, PRDM14, PRDM9, and PRMT6 (p = 0.061)  . Regardi= 0.061) . Combine= 0.061) .Breast cancer is a complex and multifactorial disease and the most common cancer in women worldwide . Currentp = 0.009). Another interesting finding was that combining chemotherapy with radiotherapy might result in a better survival rate in the high-risk subgroup of GC1. Hence, the panel of high-risk gene expression in GC1 may play a role as a promising predictive index and may also be effective in predicting treatment response to chemotherapy combined with radiotherapy in patients with TNBC. The risk estimation effects of GC1 maintained a consistent threshold in the TNBC and TNBC-LN-positive subgroups. Although no significant differences were found, the high-risk subgroup was more likely to achieve a poor PFS than the low-risk subgroup. The marginal effects of statistical significance may be because of the restricted sample size.In the present study, a serial comprehensive analysis was conducted based on hierarchical clustering results. Thirty-one genes were divided into two clusters, thereby fulfilling the optimal number. We found that the high-risk group in GC1 had a higher proportion of TNBCs than the low-risk group. Moreover, patients in the high-risk subgroup of GC1 displayed a significantly lower PFS than those in the low-risk subgroup  is a histone methyltransferase (HMT) that methylates glycine- and arginine-rich motifs (GAR motifs). PRMTs are classified into three groups according to their differences in catalytic activity: type I , type II (PRMT5 and 9), and type III (PRMT7). Next, we focused on discussing the relationship among PRMT2, PRMT8, PRMT7, and previous findings. The mRNA expression analysis of PRMT2 showed a negative median range in patients with breast cancer, and the high-risk group of GC1 reflected disputable consequences in this study. Previous studies also indicated that the role of PRMT2 in breast cancer progression remains controversial. A study suggested that spliced variants of PRMT2 are capable of binding to ERα, which further promotes EHMTRα-mediated EMT-related genes E-cadherin and Snail [PRMT2 in breast cancer. PRMT8 is an enzyme whose exact mechanism of action in breast cancer is poorly understood. Previous study indicated that PRMT8 was associated with a poor prognosis via z-score analysis, whereas the Kaplan-Meier plot showed a protective trend for breast cancer [PRMT8 expression correlated with worse survival in gastric cancer; however, increased survival was observed in patients with breast and ovarian cancer. These contrasting results may be attributed to variant-specific expression of PRMT8, including the novel PRMT8 variant 2 [PRMT8 in GC1 was negatively expressed in both the high- and low-risk subgroups. E-cadherin, an essential biomarker of EMT, is downregulated by PRMT7 at its promoter, with elevated H4R3me3 and decreased H3K4me3, H3, and H4Ac. PRMT7 interferes with E-cadherin expression through the PRMT7-YY1-HDAC3 complex, thus inducing cell migration and invasion in the MDA-MB-231 TNBC cell line [PRMT7 in the low-risk subgroup of GC1, which may indicate an insidious pathway or variations in PRMT7 enzyme activity.Protein arginine methyltransferase  in GC1, and half of them showed negative expression , whereas the rest showed positive expression  in the high-risk group. However, studies of PRDM7 and carcinogenesis are limited. Sorrentino et al. observed that PRDM7 was overexpressed in hepatocarcinoma specimens through pan-cancer reanalysis of TCGA datasets. PRDM9 has been reported to interfere with homologous recombination by binding DNA using its zinc fingers, and H3K4/36me3 occurs in surrounding nucleosomes [PRDM9 have yet to be correlated with some solid tumors, such as head and neck squamous cell carcinoma and bladder cancer [PRDM12 may act as a tumor suppressor in patients with CML [PRDM12 is upregulated in prostate and colon cancers compared to normal tissues [PRDM13 acts as an anti-proliferation regulator by upregulating INCA1, a CDK inhibitor, and ADAMTS12, thereby inhibiting cell invasion. Overexpression of PRDM13 in U87 glioma cells significantly inhibits cell migration and invasion by interacting with deleted liver cancer 1 (DLC1) and ARHGAP30 (Rho GTPase-activating protein 30) genes [PRDM13 overexpression in many cancers. These results are consistent with our study, in which PRDM13 expression was higher in the high-risk subgroup of GC1. PRDM15 could be a key regulator in diffuse large B-cell lymphoma (DLBCL), which sustained the activity of the PI3K/AKT/mTOR pathway and glycolysis [PRDM15 could also stabilize the naïve pluripotency of embryonic development by modulating the transcription of upstream regulators of WNT and MAPK-ERK signaling [PRDM14 in breast cancer, whereas low or no expression was found in non-tumorous cells. Moreover, its amplification has been correlated with more aggressive cases of breast cancer, including high-grade and distant metastases. Knockdown of PRDM14 suppressed cell growth by inducing apoptosis and increasing sensitivity to chemotherapy agents [PRDM14 mRNA expression was negative for both high- and low-risk GC1 in our study. Evidence has shown that significant PRDM14 methylation occurs in high-grade non-muscle invasive bladder, colon, and lung cancers. PRDM14′s role as a tumor suppressor—and whether its variant isoform exists—remains unknown [To date, the  repeats . Hierarcleosomes . It involeosomes ,43. Mutar cancer ,45. Prevwith CML . However tissues . PRDM13 0) genes . Howeverycolysis . In addiignaling . Previouy agents ,52,53. H unknown ,55,56.DNMT1, DNMT2, DNMT3A, DNMT3B, DNMT3L, HDAC1, HDAC2, HDAC3, PRDM7, PRDM9, PRDM12, PRDM13, PRDM14, PRDM15, PRMT1, PRMT2, PRMT5, PRMT6, PRMT7, and PRMT8, could be associated with an increased risk of poor PFS in patients with lymph node invasion. Additionally, the study suggested that the combination of chemotherapy and radiotherapy may be more effective in improving PFS compared to chemotherapy alone. We also tried to determine crosstalk between pathways with four gene families, including DNMT, HDAC, PRMT, and PRDM, which were illustrated using Ingenuity pathway analysis (IPA) systems . mRNA expression indicates the RPKM-normalized log2-transformed differential expression profiles  for the candidate genes. The mRNA expression of the candidate genes in all patients was visualized using a heatmap and the correlation between candidate genes were illustrated using a heatmap and tested using Pearson’s correlation test.k clusters was calculated to determine the optimal number of clusters for the candidate genes. Hierarchical clustering was conducted to generate dendrograms for candidate genes according to the gene similarity in mRNA expression. The greatest silhouette width was considered the optimal number of clusters to ensure the greatest dissimilarity between the identified n gene clusters. The cluster plot of candidate genes generated according to the assigned n gene clusters showed the similarity of the clustered genes in a two-dimensional distribution. Next, mRNA expression in the n gene cluster was used to dichotomize the study population into two risk estimation subgroups according to PFS status. The cutoff criteria for the two risk estimation subgroups were determined based on the distance metrics computed using a hierarchical algorithm based on the similarity of correspond mRNA expression. The computed distances were then used to establish cutoff values that separated the study population into two risk estimation subgroups, the subgroup with a higher proportion of disease progression was classified as the high-risk group, while the subgroup with a lower proportion was classified as the low-risk group.The mRNA expression levels of candidate genes were normalized into a range of 0 to 1. The average silhouette width in p-values were two-sided, and statistical significance was set at p < 0.05. All analyses were performed using the R 4.0.5 software .The baseline characteristics of all patients and the risk estimation subgroup determined by each gene cluster were summarized as frequency and percentage, and age at diagnosis was summarized by mean and standard deviation. The difference in baseline characteristic distribution between the risk estimation subgroups was estimated using an independent two-sampled t-test or chi-squared test. The mRNA expression of all candidate genes in all patients and the risk estimation subgroup were summarized using medians and ranges. The difference in mRNA expression between the risk-estimation subgroups was estimated using the Wilcoxon rank-sum test. The PFS between risk estimation subgroups in all patients, different lymph node invasion status, and treatment groups was estimated using the Kaplan–Meier method, and the survival difference between subgroups was tested using the log-rank test. All In this study, we aimed to identify an optimal predictive biomarker for TNBC. First, we searched the TCGA database to select 486 patients, and 31 candidate genes were identified. The hierarchical clustering analysis represented two clusters according to the optimal number. GC1 was significantly associated with patients with breast cancer. Second, Kaplan–Meier analysis showed shorter PFS in the high-risk group of GC1 and lymph node invasion subgroups. The trend of improved PFS in the chemotherapy combined with radiotherapy group suggests that the high risk in GC1 genes may indicate better prognosis under treatment. Herein, we propose that the high-risk group genes in GC1 may be promising predictive biomarkers and predict clinical benefits from chemotherapy combined with radiotherapy.


Myelodysplastic/myeloproliferative (MDS/MPN) neoplasms are blood disorders characterized by abnormal cell growth and development. These disorders encompass various subtypes, including chronic myelomonocytic leukemia, atypical chronic myeloid leukemia, and others. They are caused by genetic changes in different cell components and have distinct clinical features. Current treatment options mostly involve drugs that control the disease, including hypomethylating agents, ruxolitinib, lenalidomide, and venetoclax, but do not offer a cure. However, allogeneic bone marrow transplantation has the potential to cure these disorders. Several factors, such as overall health, spleen enlargement, and genetic alterations, can influence the outcome of transplantation. Future research is crucial to improving treatment approaches and patient outcomes for MDS/MPN neoplasms. This review provides an overview of the diagnosis, biology, and current and upcoming treatments, including bone marrow transplantation, for these complex blood disorders. This review will shed light on the complexities of MDS/MPN neoplasms and will inform future research for improved therapeutic strategies and patient care in the future.RUNX1), signaling molecules , splicing factors , and epigenetic regulators , as well as specific cytogenetic abnormalities . Clinical studies exploring therapeutic options for higher-risk MDS/MPN overlap syndromes mostly involve hypomethylating agents, but other treatments such as lenalidomide and targeted agents such as JAK inhibitors and inhibitors targeting PARP, histone deacetylases, and the Ras pathway are under investigation. While these treatment modalities can provide partial disease control, allogeneic bone marrow transplantation  is the only potentially curative option for patients. Important prognostic factors correlating with outcomes after allo-BMT include comorbidities, splenomegaly, karyotype alterations, and the bone marrow blasts percentage at the time of transplantation. Future research is imperative to optimizing therapeutic strategies and enhancing patient outcomes in MDS/MPN neoplasms. In this review, we summarize MDS/MPN diagnostic criteria, biology, and current and future treatment options, including bone marrow transplantation.Myelodysplastic/myeloproliferative neoplasms (MDS/MPN) are hematological disorders characterized by both proliferative and dysplastic features. According to the 2022 International Consensus Classification (ICC), MDS/MPN consists of clonal monocytosis of undetermined significance (CMUS), chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), MDS/MPN with SF3B1 mutation (MDS/MPN-T-SF3B1), MDS/MPN with ring sideroblasts and thrombocytosis not otherwise specified (MDS/MPN-RS-T-NOS), and MDS/MPN-NOS. These disorders exhibit a diverse range of genetic alterations involving various transcription factors (e.g.,  Myelodysplastic/myeloproliferative neoplasms (MDS/MPN) are a distinct group of hematological disorders characterized by overlapping features of myelodysplastic syndromes (MDS) and myeloproliferative neoplasms (MPN). Both elements are required to diagnose MDS/MPN, with the identification of the hyperproliferation of hematopoietic cells in conjunction with bone marrow dysplasia and ineffective hematopoiesis, which occasionally leads to cytopenia. In instances where cytopenia is not present, morphological analysis of the bone marrow may show dysplastic changes in one or more lineages, fulfilling the criteria for MDS/MPN diagnosis.WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues (WHO4), first released in 2008 and updated in 2016, which divided MDS/MPN into four subdivisions: chronic myelomonocytic leukemia (CMML), atypical chronic myeloid leukemia (aCML), myelodysplastic/myeloproliferative neoplasm with ringed sideroblasts and thrombocytosis (MDS/MPN-RS-T), MDS/MPN-unclassifiable (MDS/MPN-U), and Juvenile myelomonocytic leukemia (JMML) [In recent years, the classification and understanding of MDS/MPN overlap syndromes have evolved, shedding light on their complex biology and genetic abnormalities. Traditionally, the classification of MDS/MPN was based on the fourth edition of the a (JMML) . Given tThe frequency of MDS/MPN overlap syndromes varies, with CMML accounting for most cases, with an incidence ratio of 0.6 per 100,000 patients, followed by aCML  . All MDS9/L and ≥10% of the WBC) with cytopenia and the presence of <20% of blasts in the peripheral blood (PB) and bone marrow (BM) [9/L [9/L and the myelodysplastic group (MD-CMML) with lower WBC counts [CMML is defined as monocytosis (absolute monocytes ≥ 0.5 × 10row (BM) . A clonaBM) [9/L ,6. BM tyBM) [9/L . MoreoveBM) [9/L . Of noteBM) [9/L . FinallyC counts .9/L due to an increased number of neutrophils and their precursors, cytopenia, and blasts less than 20% of the composition of cells in PB and BM. Dysgranulopoiesis is present in PB, including hyposegmented and/or hypersegmented neutrophils and hypercellular BM with granulocytic proliferation and dysplasia [ACML is defined as leukocytosis 13 × 109/L and ≥10% of the WBC) with the presence or absence of cytopenia, in the presence of myeloid neoplasm-associated mutation [CMUS is a premalignant precursor to CMML defined by monocytosis (absolute monocytes ≥ 0.5 × 10mutation . No morpmutation . In the mutation .SF3B1 mutation (MDS/MPN-T-SF3B1) is defined as thrombocytosis  with anemia with minimal blasts (<1% in PB and <5% in BM) [S3FB1 mutation is necessary for diagnosis, with or without other myeloid neoplasm-associated mutations or abnormal cytogenetics, but always without BCR-ABL1 translocation or other genetic abnormalities of myeloid/lymphoid neoplasms with eosinophilia and tyrosine kinase gene fusion [MDS/MPN with thrombocytosis and 9/L) with minimal blasts (<1% in PB and <5% in BM) and anemia with erythroid-lineage dysplasia and >15% ring sideroblasts [S3FB1 mutation, BCR-ABL1 translocation, or other genetic abnormalities of myeloid/lymphoid neoplasms with eosinophilia and tyrosine kinase gene fusion should be identified. The presence of other MDS, MPN, or MDS/MPN needs to be excluded [MDS/MPN-RS-T, NOS is defined as thrombocytosis  and blasts in PB and BM < 20% . ClonaliTET2 and SRSF2 mutations are the most common founder mutations in CMML, while ASXL1/SETBP1 mutations co-occur frequently in aCML [ASXL1, EZH2, RUNX1, SETBP1, NRAS, STAG2), which were associated with a higher risk of AML transformation and worse survival [The genomic profile of patients with MDS/MPN defines the disease biology and strongly affects patients’ outcomes . Palomo  in aCML . The aut in aCML . Finally in aCML . Our grosurvival .TET2 and ASXL1; RNA splicing, including SRSF2; cell signaling, such as CBL and NRAS; and transcription and nucleosome assembly, including RUNX1 and SETBP1 [SRSF2, TET2, and ASXL1 are reported to be the most frequently mutated of these genes, present in approximately 40% of patients with CMML [TET2 is thought to be the initial driver mutation responsible for monocytosis, but it has not been shown to impact OS or LFS in CMML [ASXL1, along with DNMT3A, RUNX1, SETBP1, NRAS, KRAS, CBL, and JAK2, is associated with increased proliferation, dysplasia, and progression to AML [ASXL1 mutations are the only mutations independently and consistently associated with poor prognosis [Somatic genetic mutations are present in over 90% of patients with CMML, can aid in the confirmation of diagnosis, are predictive of the disease course, and present opportunities for developing novel therapeutics ,9. Some d SETBP1 ,15,16,17ith CMML . TET2 is in CMML ,19. The n to AML ,21,22. Nrognosis ,23,24,25rognosis .JAK2, CALR, and MPL, and the presence of SETBP1 and ASXL1 mutations can provide additional support for aCML diagnosis, according to ICC guidelines [ASXL1 mutation, found in the ancestral clone in 79% of cases [ASXL1 (28%), TET2 (16%), NRAS (16%), SETBP1 (12%), and RUNX1 (12%) as the most prevalent mutations [TET2, NRAS, and PTPN11 mutations, along with the presence of more than three mutations, were found to adversely impact survival in univariate analysis, while ASXL1, SETBP1, and ETNK1 were not found to impact prognosis [TET2 was the only mutation that retained association with a worse prognosis outcome in multivariate analysis [In aCML, the absence of MPN-associated driver mutations, such as idelines ,27. Palos [ASXL1 %, TET2  [p < 0.001), respectively [ASXL1 mutations were detected in 37% of patients with abnormal karyotypes, while SP3B1 mutations were detected in 46% of patients with normal karyotypes [Cytogenic abnormalities are present in approximately 30% of all patients with CMML ,26. An a= 0.001) . In this= 0.001) . Stratifectively . The Mayectively . High-riectively . ASXL1 mryotypes .In an analysis of 367 patients with different subsets of MDS/MPN, aCML and MDS/MPN-NOS were associated with the highest genomic instability: 42% and 47% of patients had chromosomal abnormalities, respectively . An analTo date, the mainstay of first-line chemotherapy for MDS/MPN includes hypomethylating agents (HMA). HMA, including 5-azacitidine (AZA) and decitabine (DAC), decrease oncogenesis-related DNA methylation by irreversibly binding to DNA methyltransferase and can also cause direct DNA damage ,35,36,37Since then, clinical trials of HMA in CMML-specific populations have been performed. A meta-analysis of fourteen pooled studies, including 600 patients with CMML, identified an estimated ORR that was very similar between the DAC and AZA groups, but with CR rates slightly higher in patients treated with DAC (23% in DAC vs. 10% in AZA) . DrummonHMA are also used as a first-line treatment in non-CMML MDS/MPN, with their rationale for use drawn mainly from CMML studies. Case reports have explored the use of DAC in aCML, with pooled data identifying a complete hematologic remission (CHR) in seven of eight patients, with two patients being bridged to transplant ,50,51,522 had similar DNA methylation, efficacy, and safety in the initial two cycles [2 [HMA must be given IV due to cytidine deaminase, an enzyme that breaks down DAC and AZA in the small intestine and liver, reducing HMA effectiveness when taken orally. This poses challenges for patients limited by transportation and can reduce quality of life due to the excess time spent in hospitalization receiving care . Cedazuro cycles . A phaseycles [2 .Other enhancements to HMA therapy include guadecitabine, a next-generation hypomethylating agent with prolonged metabolic activity. Guadecitabine has been tolerated in a study of high-risk myelodysplastic syndromes including 22 CMML patients . The ORRSETBP1, RUNX1, or EZH2 genes is associated with a worse response to HMA and inferior OS in a cohort of patients with MDS/MPN treated with AZA or DAC [ASXL1 gene mutations predicted lower ORR to HMA (OR = 0.85), while TET2mut/ASXL1wt was found to be predictive of a higher CR rate (odds ratio (OR) = 1.18) and a better OS in multivariate analysis [RUNX1 mutation was similarly associated with a worse OS, along with mutation in the CBL gene and higher WBCs [Overall, marginal response rates, the development of HMA resistance, and a lack of alterations in the disease course warrant the investigation of novel agents in MDS/MPN . SimilarA or DAC . Duchmananalysis . The RUNher WBCs ,64. FurtRuxolitinib is a JAK1/2 inhibitor with approved use in polycythemia vera, myelofibrosis, and acute and chronic graft-versus-host disease. A preclinical study of CMML found that the pro-neoplastic JAK1/2 pathway can be induced by the granulocyte-macrophage-colony-stimulating factor (GM-CSF) in CMML primary samples and can be successfully targeted by JAK inhibitors, including ruxolitinib . SeveralCSF3R) in aCML has prompted the consideration of ruxolitinib to halt the aberrant signaling through JAK inhibition [CSF3R, causes a lethal myeloproliferative disease in mice, and the degree of splenomegaly and leukocytosis has been demonstrated to be reversed by ruxolitinib [CSF3R-T618I, hydroxyurea-refractory aCML, the use of ruxolitinib led to a reduction in constitutional symptoms, leucocytosis, and the spleen size, as well as an improvement in anemia and thrombocytopenia [CSF3R mutation; two patients were non-responders, while four patients reached the end of cycle six of ruxolitinib treatment [CSF3R T618I or JAK2V617F mutations [The identification of mutations of the colony-stimulating factor 3 , small lymphocytic lymphoma (SLL), and AML patients who are not eligible for standard chemotherapy. Montalban-Bravo et al. retrospectively investigated the activity of venetoclax-based therapy in 27 CMML patients and 26 patients with AML-MRC with anteceding CMML . Ventoclp = 0.2) as well as the ORR  at 12 months for the entire cohort [Lenalidomide and thalidomide are immune-modulatory agents with several mechanisms of action, including antiangiogenic properties, the repression of IL-6 production, and the activation of apoptotic pathways . Lenalide cohort . The CMMe cohort .Lenalidomide has also been used in patients with MDS/MPN-RS-T . Based oBRCA1 and BRCA2, which prevent effective homologous recombination, can accumulate mutations, leading to reliance on PARP1 for DNA repair and survival [PARP inhibitors like veliparib are FDA-approved breast and ovarian cancer treatments and have demonstrated preclinical efficacy in primary MPN samples . PARP ensurvival ,91. Pratsurvival . This prsurvival . A totalsurvival . Patientsurvival . Six patsurvival .9 to 110 × 109 for 22 weeks [Trametinib is a mitogen-activated protein kinase 1 MEK1/MEK2 inhibitor that inhibits ERK phosphorylation in the RAS pathway and has demonstrated a reduction in NRAS-mutated AML proliferation in pre-clinical studies . Approxi22 weeks . A rando22 weeks ,98. No d22 weeks . In a ca22 weeks . RAS patHistone deacetylases inhibitors (HDACIs), including panobinostat (PAN), entinostat, and vorinostat, can lead to an increased expression of genes, inducing apoptosis, cellular differentiation, and cell-cycle arrest, that are aberrantly suppressed in oncogenesis . HDACIs In contrast, a phase II randomized trial  investigated entinostat with AZA, which resulted in a median OS of 22.2 months for treatment with AZA alone as opposed to 14.7 months with AZA and entinostat . The comRas/CBL/KAK2 signaling mutations [Other targeted therapies have been evaluated in a few clinical studies of MDS/MPN—mainly, CMML. Lenzilumab is a monoclonal antibody against GM-CSF, which induces apoptosis in cells with high GM-CSF receptor expression, which is a protein often found on myeloid progenitor cells ,108. As utations . A phaseutations . Prelimiutations .PANTHER, a randomized phase III clinical trial of pevonedistat and AZA vs. AZA monotherapy, 27 CMML patients demonstrated a similar OS and EFS between arms, with an ORR of 44% in the pevonedistat plus AZA arm compared to 36% in the AZA monotherapy arm [Pevonedistat is a small molecule inhibitor of the neural precursor cell (NEDD8)-activating enzyme leading to downstream protein ubiquitination . Pevonedrapy arm .Omacetaxine mepesuccinate (OM) is a semi-synthetic form of hemoharringtonine (HHT), a plant alkaloid with historical significance as an antineoplastic therapy in China . HHT prePEG-IFN-alpha was utilized in CML before imatinib development and may have some limited utility in MDS/MPN, including in aCML and MDS/MPN-RS-T. In a retrospective analysis of aCML patients, only one of seven patients treated with IFN-alpha responded  . In a phThe ABNL-MARRO study is an international collaboration that enables a framework for creating clinical trials for the heterogenous MDS/MPN population. The ABNL-MARRO study is currently enrolling patients in a trial investigating itacitinib, a selective JAK1 inhibitor with an oral combination of DAC and the cytidine deaminase inhibitor, cedazuridine, as the first investigation in an international clinical trial .The only therapeutic modality that is potentially curative for patients with MDS/MPN is allogeneic bone marrow transplantation . To date, many studies have explored the clinical outcomes of allo-BMT for patients with MDS/MPN. Most studies have evaluated a small subpopulation of CMML, in conjunction with a larger pool of MDS or AML patients and, to a lesser extent, by itself. One of the first retrospective studies evaluating allo-BMT outcomes in 21 patients with CMML was performed by Zang et al. in the Fred Hutchinson Cancer Research Center, reporting a 3-year disease-free survival (DFS) and relapse rate (RR) of 39% and 25%, respectively . A shortp = 0.01) [9/L), and neutropenia (ANC < 1.5 × 109/L) at the time of transplant [ASXL1, SRSF2, and TET2 mutations had a prognostic impact [CMML transplant score) that accounts for ASXL1 and/or NRAS mutations (four points), a percentage of BM blasts > 2% (four points), and a comorbidity index (one point for eight different comorbidities), which cumulates in five risk groups with significant differences in survival and non-relapse mortality [Additional clinical studies have evaluated clinical and molecular prognostic factors of allo-BMT in CMML. A French study analyzed outcomes in 73 patients with CMML who received allo-BMT between 1992 and 2002 . Transpl = 0.01) . A studyansplant . Better ansplant . Despitec impact . Gagelmaortality .A European Group for Blood and Marrow Transplantation study retrospectively evaluated transplantation outcomes in 513 patients with CMML and reported 4-year NRM, RR, DFS, and OS values of 41%, 32%, 27%, and 33%, respectively . The onlAnother multicenter study from Liu et al., which utilized data from the Center for International Blood and Marrow Transplant Research (CIBMTR), investigated prognostic factors associated with transplant outcomes in 209 adult patients with CMML receiving allo-BMT . A lowerMortality after transplant is mainly caused by transplant-related mortality (TRM) or the relapse and progression of disease. The TRM in the first 100 days post-transplant is primarily driven by severe acute GVHD, with different reports finding this percentage to range from 25 to 41% . OptionsThe utilization of allo-BMT for aCML was first described in a case report . In 2004Few studies have exclusively evaluated the clinical outcomes of allo-BMT in MDS/MPN-NOS. A collaborative analysis between the Mayo Clinic and Moffitt investigated outcomes in MDS-MPN-NOS patients, identifying 63% who remained alive and disease-free at a median follow-up time of 61 months . A JapanEZH2/RUNX1/SETBP1 genes [ASXL1, CBL, TET2, and NRAS used next-generation sequencing as a predictor for relapse after transplant in MDS/MPN [CSF3R T618I mutation, and the detection of MRD with PCR for WT1 or flow cytometry can be used as predictors of relapse post-transplant [Despite the uniqueness of each MDS/MPN entity, studies have reported combined outcomes, often because primary analysis shows no differences after stratifying for each neoplasm. A recent collaborative analysis of 15 North American centers evaluated the haploidentical BMT with PTCy in 120 patients with MDS/MPN and reported a 3-year NRM, PFS, relapse, and OS, of 25%, 48%, 27%, and 56%, respectively . Severe P1 genes . A mutat MDS/MPN . Patient MDS/MPN . These mansplant ,155.Another area of research lies in the conditioning regimens and bridging therapies before allo-HSCT. Traditionally, myeloablative conditioning (MAC) involving cytotoxic agents, like busulfan with cyclophosphamide and/or total body irradiation (TBI) ≥ 5 Gy, is preferred in young patients. In contrast, reduced-intensity conditioning (RIC) regimens are usually preferred in older patients or patients with comorbidities. A study comparing MAC and RIC in patients with MDS/MPN syndromes showed relapse in all patients (5/10) receiving RIC but no relapse in those pre-treated with MAC (5/10), underscoring the survival benefit associated with more intense myeloablation . HoweverMDS/MPN overlap syndromes represent a unique and challenging group of hematological disorders. The understanding of these diseases has significantly advanced, particularly regarding their underlying biology and genetic abnormalities, providing insights into the pathogenesis and potential therapeutic targets. Current treatment approaches encompass a multimodal strategy, including disease-modifying agents, targeted therapies, and supportive care, with allo-BMT being the only potentially curative option for eligible patients. However, challenges remain in optimizing patient selection, refining transplant techniques, and minimizing transplant-related complications. Future clinical research directions on MDS/MPN overlap syndromes should focus on several key areas. To guide personalized therapy decisions, efforts should be made to develop risk-adapted treatment algorithms based on comprehensive genetic profiling, disease characteristics, and patient factors. Refining allo-BMT protocols, such as RIC regimens and improved GVHD prophylaxis strategies, holds promise in reducing treatment-related morbidity and mortality. Moreover, identifying reliable biomarkers for disease monitoring, response assessment, and the early detection of relapse would greatly enhance clinical management. Ultimately, collaborative research endeavors and international registries are vital, as they facilitate the accumulation of larger patient cohorts, leading to a better understanding of disease heterogeneity and refining prognostic models.


The two members of the mammalian STIM family, STIM1 and STIM2, are transmembrane proteins that act as Ca2+ sensors in the endoplasmic reticulum (ER) and, upon Ca2+ store discharge, interact with and activate the Orai/CRACs in the plasma membrane. Dysregulation of Ca2+ signaling leads to the pathogenesis of a variety of human diseases, including neurodegenerative disorders, cardiovascular diseases, cancer, and immune disorders. Therefore, understanding the mechanisms underlying Ca2+ signaling pathways is crucial for developing therapeutic strategies targeting these diseases. This review focuses on several rare conditions associated with STIM1 mutations that lead to either gain- or loss-of-function, characterized by myopathy, hematological and immunological disorders, among others, and due to abnormal activation of CRACs. In addition, we summarize the current evidence concerning STIM2 allele duplication and deletion associated with language, intellectual, and developmental delay, recurrent pulmonary infections, microcephaly, facial dimorphism, limb anomalies, hypogonadism, and congenital heart defects.The STIM family of proteins plays a crucial role in a plethora of cellular functions through the regulation of store-operated Ca Ca2+ intervenes in a wide range of cellular processes, including cell signaling, gene expression, fertilization, muscle contraction, and neurotransmitter release. The precise regulation of intracellular Ca2+ levels is essential for maintaining cellular homeostasis and proper behavior of various physiological processes. Ca2+ acts as a versatile signaling molecule due to its ability to rapidly change its cytosolic concentration in response to extracellular stimuli. This dynamic regulation requires the synergistic action of various Ca2+ sensors, ion channels, pumps, and transporters that control Ca2+ efflux and influx across intracellular compartments and the plasma membrane (PM) ER. Upon store depletion, STIM proteins undergo conformational changes and translocate to specialized regions called puncta or ER-PM junctions ER, which is in the range of 100 to 800 μM. The initial studies by nuclear magnetic resonance revealed that the N-terminal portion of STIM proteins contains paired EF-hand structures that form a hydrophobic pocket, which is stabilized by the sterile α-motif (SAM) domain  and results in a compact globular structure. Whereas the canonical EF-hand domain  binds Ca2+, with the half-maximum activation occurring at 200 μM (STIM1) and 400 μM (STIM2), the second non-canonical or hidden EF-hand motif  stabilizes the canonical EF-hand, supported by the action of the SAM domain [2+ binds, which encompasses 11 residues  of the EF-hand. This pocket is secured by L195 and L199 of the SAM domain of STIM1. The arrangement of negatively charged residues on the surface of this globular motif includes D77, D78, D82, D84, E86, E87, D89, E90, E94, and D95 from the canonical EF-hand; E111, D112, E118, and E128 from the non-canonical EF-hand; and D135, E136, and D196 from the SAM domain. These negatively charged residues likely prevent intermolecular interactions with other N-terminal STIM1 domains [STIM1 and STIM2 can detect changes in [CaM domain ,28,29,30The luminal and cytosolic domains of STIM proteins are divided by a single transmembrane helix that spans the ER membrane  . This TMThe C-terminus of STIM proteins is characterized by three coiled-coil (CC) domains, namely, CC1, CC2, and CC3 . These CC domains are vital for the interaction with and gating of Orai proteins, and therefore, SOCE. The function of the CC domains was independently identified almost simultaneously by three groups, who named it as the Orai-activating small fragment (OASF), composed of CC1, CC2, and CC3 , CRAC-acAs we discuss in the following sections, point mutations or polymorphisms in any of the mentioned motifs lead to STIM1 abnormal function gain/loss, hence, altered SOCE, and subsequently, pathophysiological conditions.2+ influx upon depletion of the intracellular Ca2+ stores. Among the most relevant disorders associated with gain-of-function mutations of STIM1 are Stormorken syndrome, York platelet syndrome (YPS), and tubular aggregate myopathy (TAM).As mentioned above, STIM1 is involved in a plethora of cellular functions and a number of disorders have been associated with STIM1 mutations that lead to either gain- or loss-of-function resulting in constitutive activation of CRACs or deficient Ca2+ concentration, which is significantly elevated upon stimulation with physiological agonists, leading to exposure of phosphatidylserine at the plasma membrane [2+ concentration and reduced or absent SOCE upon Ca2+ store depletion, which has been attributed to near-maximal Orai1 activation at resting conditions [The Stormorken syndrome is a rare autosomal-dominant genetic condition identified by Helge Stormorken in 1984 in a patient suffering from long-lasting bleeding after a minor accident . This symembrane ,48, Stornditions . Therefonditions . Paradoxnditions ,50. Thisnditions .2+ concentrations. This mechanism depends on the formation of a coiled-coil clamp involving the CC1 and CC3 domains [2+ store depletion [Almost three decades after the identification of this condition, the molecular basis of the syndrome was identified as a gain-of-function mutation located in the α2 segment of the STIM1 coiled-coil region 1 (CC1), the STIM1-R304W ,49,52 T. The mis domains ,53. Althepletion . Consistepletion . In addiepletion   leading to a STIM1 K365N amino acid substitution  Table 1Table 1. The Stormorken syndrome clinically overlaps with TAM, a progressive muscle disorder associated with weakness, cramps, myalgia, increased creatine kinase levels, and accumulation of packed membrane tubules containing sarcoplasmic reticulum proteins. These tubules are observed by electron microscopy as straight, single- or double-walled tubules that are highly ordered and aligned in parallel in a longitudinal muscle section ,57. Furt2+ concentration [2+ concentration but the amplitude of SOCE was significantly higher in TAM vs. control myoblasts [TAM is mediated by several autosomal dominant mutations in the EF-hand domain of STIM1, which result in constitutive CRAC activation. Initially, four activating STIM1 mutations were identified through Sanger sequencing of the STIM1 coding exons and exon–intron boundaries in families of patients with common histological features or by exome sequencing of TAM patients: H72Q, D84G, H109N, and H109R ; Table 1ntration . Two morntration  and STIMntration . In contyoblasts .2+-handling protein calsequestrin-1. Three missense mutations in the calsequestrin-1 gene have been identified in patients with a mild phenotype associated with tubular aggregate myopathy, including D44N, G103D, and I385T [TAM might also be caused by mutations in other proteins, including Orai1 and the reticular Cand I385T . Furthernd I385T .2+ and phosphorous, in a ratio that resembles that observed in normal platelet dense bodies, as well as peroxidase and acid phosphatase activity, like primary lysosomes. As the number of lysosomes was reduced in YPS platelets, it has been reported that both opaque and target organelles are either abnormal lysosomes or fused with lysosomes during their development [The York platelet syndrome was first reported in a series of articles published in 2003 describing ultrastructural abnormalities found in platelets from a woman and her male child who manifested life-long thrombocytopenia although functionally normal in aggregation ,66,67,68elopment . In addielopment ,70.STIM1 variants, c.343A>T and c.910C>T, encoding for the STIM1 mutations I115F and R304W, respectively [Whole exome sequencing from YPS patients revealed two ectively ,70  sequence [2+ concentration when expressed in HEK293 cells [A recent study  has reposequence ) and R74sequence . Functiosequence . FurtherThe aberrant SOCE mediated by the aforementioned gain-of-function STIM1 mutations, as well as some gain-of-function Orai1 variants, with the exception of the R304Q, has been recently reported to be sensitive to two structurally unrelated SOCE modulators, CIC-37, a compound developed from a class of pyrazole-bearing molecule, and CIC-39, a biphenyl-triazole developed from Synta66 . If thesCRACs are formed by the heterogeneous association of Orai and STIM family members. Twenty years ago, the study of STIM1 and Orai1 loss-of-function mutations allowed the identification of the molecular mediator of SOCE . Indeed,STIM1 gene. Due to the appearance of a premature termination codon, this frameshift produces a truncated STIM1 mutant that ends at the beginning of the SAM domain. The STIM1 E128fs*9 patients’ derived fibroblasts showed markedly reduced STIM1 mRNA levels and an undetectable STIM1 protein expression, making these cells unable to mediate SOCE [Up to date, six autosomal recessive STIM1 loss-of-function mutations have been described . STIM1 Eate SOCE . Patientate SOCE . This muate SOCE . In patiate SOCE . Based o2+ entry observed in T cells derived from STIM1 P165Q patients might support immune responses, increasing their lifespan compared to other CRAC channelopathy patients. Furthermore, this hypomorphic mutation has been associated with additional inflammatory disorders, such as psoriasis and colitis, that were not previously associated with other STIM1 mutations.CRAC channelopathy derived from STIM1 P165Q mutation shows a unique phenotypic trait. This mutation reduced STIM1 expression and function but might still allow for residual SOCE . Althoug2+ channels. STIM1 R429C mutation was found in two siblings born to consanguineous parents. A younger sister died from sepsis at the age of 21 months, while the older presented dental enamel defect, anhidrosis, generalized eczema, mild muscular hypotonia, light-insensitive pupils, primary enuresis, and combined immunodeficiency. Interestingly, STIM1 R429C mutation did not alter protein expression but impaired SOCE in regulatory and CD3+ T cells [2+ stores [Two loss-of-function mutations have been described within the SOAR/CAD domain of STIM1. As described above, this sequence plays a crucial role in the oligomerization of STIM molecule and the activation of store-operated Ca T cells . A study T cells . Wang et T cells . Like ST+ stores . The STI+ stores . The lat+ stores .A novel STIM1 loss-of-function mutation was identified in two patients affected with muscle weakness, dysmorphic facies, hypoplastic patellae, dental abnormalities, and hyperelasticity. This phenotype, which only presents mild immune deficiencies, was caused by a guanine insertion after position 685 in the coding sequence of STIM1, a frameshift that changes phenylalanine 229 to leucine and induces a premature termination codon. As observed in other STIM1 null mutations, cells derived from patients carrying STIM1 F229L mutation show reduced STIM1 mRNA levels and an indetectable protein expression . Howeverhttps://www.ensembl.org, accessed on 20 June 2023). There are no more than 40 cases reporting interstitial deletions or duplications on the proximal short arm of chromosome 4 and few involving alterations in STIM2 alleles. The most prevalent phenotype associated with such chromosomal alterations is intellectual disability, presenting other clinical features to a minor degree, such as physical and facial dimorphism or congenital heart diseases [The STIM2 gene is located within the p15.2 band of the short arm of chromosome 4 and adjacent to the p15.1 band (diseases ,88. For diseases . This trdiseases . A good diseases . STIM2 mdiseases . Regardidiseases , and thadiseases , it is tdiseases . MoreoveTwo studies involving STIM2 allele deletion and haploinsufficiency were reported. The first study performed genetic analyses on a 3-year-old proband displaying physical and intellectual delay, microcephaly, diverse facial anomalies, anxiety, frequent laughing, and hypermobility of the elbows. A deletion of a region of 3.4 Mb in size was found at 4p15.2-p14, generating haploinsufficiency in STIM2 and 15 additional genes . From th2+ homeostasis, more precisely, the activation of Ca2+ influx through CRACs. Because of the key physiological role of STIM1, several point mutations associated with STIM1 gain-of-function lead to multi-systemic disorders with overlapping features, including platelet dysfunction and thrombocytopenia, bleeding diathesis, or congenital myopathy. Although the Stormorken syndrome, TAM, and YPS are rear conditions with a well-recognized pathogeny associated with mutations in the N-terminal STIM1 region, certain STIM1 polymorphisms lead to diffuse phenotypes not clearly associated with them that deserve further studies. Furthermore, several STIM1 loss-of-function mutations have been identified which lead to moderate or complete impairment of CRAC activation and are associated with immune deficiency, impaired enamel formation, or nail dysplasia, among other disorders. Several disorders have also been associated with STIM2 allele duplication or deletion, leading to STIM2 overexpression or absence, which are characterized by intellectual delay, microcephaly, physical and facial dimorphism, or congenital heart diseases. Characterizing and understanding the genetic and molecular mechanisms underlying STIM protein function are crucial for developing therapeutic strategies targeting these diseases.STIM proteins, mainly STIM1, play a crucial role in cell biology as a result of the regulation of intracellular Ca


Circoviridae family within the genus Circovirus, poses a significant economic risk to the global swine industry. PCV2, which has nine identified genotypes (a–i), has emerged as the predominant genotype worldwide, particularly PCV2d. PCV2 has been commonly found in both domestic pigs and wild boars, and sporadically in non-porcine animals. The virus spreads among swine populations through horizontal and vertical transmission routes. Despite the availability of commercial vaccines for controlling porcine circovirus infections and associated diseases, the continuous genotypic shifts from a to b, and subsequently from b to d, have maintained PCV2 as a significant pathogen with substantial economic implications. This review aims to provide an updated understanding of the biology, genetic variation, distribution, and preventive strategies concerning porcine circoviruses and their associated diseases in swine.Porcine circovirus (PCV), a member of the  Circovirus and family Circoviridae, and is responsible for porcine circovirus virus-associated disease (PCVAD), which includes a number of diseases such as post-weaning multisystemic wasting syndrome (PMWS), porcine dermatitis and nephropathy syndrome (PDNS), granulomatous enteritis, porcine respiratory disease complex, reproductive failure, and acute pulmonary edema [Porcine circovirus (PCV), a ubiquitous viral pathogen of pigs, belongs to the genus ry edema ,2. PCV wry edema . Later sry edema . Subsequry edema ,6, Northry edema ,8. Sequery edema . To diffry edema . Despitery edema . At presry edema . PCVs arry edema ,14. Althry edema ,16. In tPCV is a small, non-enveloped, icosahedral, single-stranded circular DNA virus  with a dThe genomic DNA sequence homology between PCV1 and PCV2 ranges from 68% to 76% ,22. BothThere are two major ORFs in PCV, namely ORF1 and ORF2, which are oriented in the antisense direction . The ORFORF3 is the most variable open reading frame between PCV1 and PCV2, with only 61.5% amino acid sequence identity . It is eThe genome of PCV3 has at least three ORFs, ORF1, ORF2, and ORF3. ORF1 is oriented in the opposite direction to that of ORF2 and encodes a single 296–297 aa replicase (Rep) protein that shares 55% amino acid sequence similarity with PCV2 proteins . ORF2 enThe genome of PCV4 contains twelve predicted ORFs including two major ORFs, ORF1 and ORF2. ORF1 encodes a replicase protein of 296 aa that shares 48.1% and 47.2% amino acid sequence identity with PCV1 and PCV2, respectively. ORF2 encodes a capsid (Cap) protein of 228 aa that shows 43.1%, 45%, and 24.5% sequence identity with PCV1, PCV2, and PCV3, respectively .−3 substitutions/site/year), like that of the RNA viruses [According to a new genotyping methodology, PCV2 has eight distinct genotypes, PCV2a to PCV2h . Recentl viruses . Among n viruses ,43. The Furthermore, PCV2a has five clusters (2A–2E), PCV2b has three clusters (1A-1C), and PCV2d has two sub-genotypes  ,45. PCV2The higher genetic variability within PCV2a compared to PCV2b suggests that PCV2a might be older genotype relative to PCV2b . PhylogePCV3, which was initially reported in the US in 2016 , has alsPCV4 was first reported in 2019 in Hunan Province, China , and in On the basis of ORF1, ORF2, and complete genome sequences, all PCV4 strains can be categorized into two major genotypes (PCV4a and PCV4b) . PCV4 isThe phylogenetic tree was constructed using 13 complete ORF2 sequences of selective representatives of different PCVs  obtained from GenBank. The tree was generated using the p-distance model-based neighbor-joining (NJ) method and bootstrapped at 1000 replications. Branches with a bootstrap vale of less than 70% were collapsed.Among four PCVs discovered so far, PCV1 is considered as non-pathogenic, while the other three PCVs, i.e., PCV2, PCV3, and PCV4, are known to be pathogenic. PCV1, PCV2, and PCV3 are distributed throughout the global pig population, while the distribution of PCV4 is yet to be known . PCV2 isPCV2-LD is equivalent to PCV2-associated porcine respiratory disease complex (PRDC) . PCV2, aPCV3 has been detected in pigs showing the symptoms of systemic disease, reproductive disease , PDNS 2,105), reInfection with PCV4 shows the symptoms of respiratory disease, enteric disease, PDNS, and PMWS ,122. ThePorcine parvovirus [Porcine reproductive and respiratory syndrome virus [Swine Hepatitis E virus [Swine influenza virus, Mycoplasma hyopneumoniae [E. coli [Salmonella spp. [M. hyopneumoniae (35.5%), bacterial septicemia pathogens (14%), bacterial pneumonia pathogens (7.6%), and SIV (5.4%) [M. hyopneumoniae as a co-infecting agent revealed the same results [Although PCV2 is the primary pathogen of PCVAD, it is alone not sufficient to cause clinical disease. Several other infectious co-factors and non-infectious conditions are crucial for the clinical expression of PCV2 infection . PCV2 inrvovirus , Porcineme virus , Swine H E virus , Swine ieumoniae , E. coli[E. coli , and Sallla spp. . A retroV (5.4%) . MoreoveV (5.4%) ,133. SucV (5.4%) . SeveralV (5.4%) ,135. DifV (5.4%) ,136. HowV (5.4%) . EvidencV (5.4%) . A studyV (5.4%) . HoweverV (5.4%) ,141. NonV (5.4%) . Another results . Such co results ,143. The results . Mutatio results . Mutatio results . Several results ,147,148.While PCV2 infections are widespread, the prevalence of PCVAD is relatively lower . ConsequThe level of maternal antibodies (MDAs) could play a critical role in determining the host’s resistance to PCV2 infection. A higher level of maternal antibodies results in increased resistance to viral infection in piglets . The levE. coli and atrophic rhinitis would act as protective factors [Epidemiological studies have shown that on-farm management factors, housing conditions, vaccination schedules, biosecurity, hygiene, and husbandry practices are strongly associated with the development of PCVAD . Large p factors ,157. The factors .Mycoplasma hyopneumoniae (M. hyopneumoniae) bacterin containing a mineral oil adjuvant exhibited PMWS after vaccination. Clinical signs were developed only in forty percent (40%) of vaccinated piglets [Mycoplasma hyopneumoniae vaccine at 1 week of age and challenged with PCV2 at 2, 7, and 12 weeks of age; the AGE12 group of pigs had higher levels of viremia compared to nonvaccinated pigs. However, this effect was not observed in younger pigs (AGE2 and AGE7) [M. hyopneumoniae vaccine along with oil-in-water adjuvant, compared to the groups treated with the same vaccine but in conjunction with different adjuvants, i.e., aqueous-carbopol and aluminum hydroxide adjuvants [Vaccines utilized for co-infecting pathogens or adjuvants utilized along with those vaccines may also affect the outcome of PCV-2 infection. In an experimental study, gnotobiotic piglets vaccinated with a commercial  piglets . Similar piglets . The age piglets . For exand AGE7) . Althougnd AGE7) . Furtherdjuvants . Many stdjuvants ,165,166.djuvants ,168.Musca domestica), stable fly  and mosquito (Culex sp.) suggested that these insects may act as a mechanical vector of PCV2 [Both horizontal (among pigs of the same generation) and vertical (mother to offspring) modes of PCV2 transmissions have been reported. However, the horizontal mode of transmission is very frequent. A recent study indicated that direct contact between pigs is the most efficient means of PCV2 transmission . However of PCV2 ,194,195. of PCV2 , pig to  of PCV2 , pig to  of PCV2 , pig to  of PCV2 , and pig of PCV2 .Like PCV2, PCV3 can be transmitted both horizontally and vertically . The priAs PCV4 is a very recently identified PCV, details about its transmission are not available. The virus has been detected in tissue specimens of respiratory, renal, digestive, circulatory, and lymphatic systems, as well as from aborted fetuses ,78,123. The diagnosis of PCVAD relies on three criteria: evaluating clinical signs, examining microscopic lesions in affected organs such as lymphoid tissue, lungs, liver, heart, kidney, and intestine, and detecting the PCV2 antigen or its DNA in lesions . When a Herd diagnosis as compared to diagnosis of a single pig is critical for the prevention and control of PCVAD. Such herd diagnosis can be conducted on the basis of two criteria: (i) a significant increase in postweaning mortality associated with clinical signs compatible with PMWS; and (ii) individual diagnosis of PMWS in at least 1 out of 3–5 necropsied pigs . FurtherCurrently, common techniques used for the detection of PCV2 and PCV3 antigens or nucleic acids include immunohistochemistry (IHC), in situ hybridization (ISH), and PCR ,211,212.PCR can also be used for the detection of PCV2 in infected tissues. PCR is fast, has higher sensitivity as compared to IHC and ISH, and can even be performed on live samples . ConventSerological techniques such as indirect immunofluorescence assay (IIFA) ,236 and The detection of newly identified PCV4 has also been undertaken by conventional PCR , SYBR GrThe complete life cycle of PCV is yet to be explored. Due to their small genome size and limited coding capacity, the life cycle of PCV relies on host factors . The sucThe ORF1 of the PCV2 genome encodes two differently spliced protein products, Rep and Rep′, which play important roles in DNA replication. Each of these two proteins has N-terminal nuclear localization signals (NLSs), of which NLS1 and NLS2 regulate nuclear import from cytoplasm for the purpose of replication, whereas NLS3 enhances the import mechanism . Both ReThe capsid protein (Cap) encoded by ORF2, apart from its structural role in encapsidation, is also crucial both for cytoplasmic transport and replication of PCVs. Cap protein facilitates circovirus trafficking through its interaction with the cytoplasmic dynein IC1 subunit and induction of acetylation of microtubule α-tubulin . LikewisMany viruses use host autophagy for enhancing their replication. PCV2 could also trigger autophagosome formation and promote autophagic flux in PK-15 cells to enhance its replication . The proThe knowledge on the life cycle of PCV3 is very limited. It is very difficult to isolate and propagate PCV3 in vitro, which makes it difficult to gain scientific information about its infection cycle, pathogenesis, and immunological response. Initially, several attempts to isolate and multiply PCV3 in ST and PK-15 cells failed ,271. ThePCV4 is a newly identified PCV genotype, and detailed information about its life cycle, including its interactions with host cells, way of entry, etc., are currently unknown. Recent studies demonstrated that NPM1 and DEAD-box RNA helicase 21 (DDX21) interact with the N-terminal nucleolar localization signal (NoLS) of Cap protein of PCV4 and play a crucial role in the replication and assembly of viral particles by modulating the translocation of the viral Cap protein from the cytoplasm to the nucleolus of infected cells ,276.®) or inactivated chimeric PCV1-2a virus (FosteraTM PCV) or subunit proteins of PCV2a capsid  [Vaccination is considered to be one of the most effective ways of controlling PCVAD . Howevernt® PCV) . All vacnt® PCV) . In effent® PCV) . ADWG want® PCV) . Vaccinant® PCV)  and numbnt® PCV) . Vaccinent® PCV) . The timnt® PCV) . Howevernt® PCV) . Thus, fnt® PCV) ,287,288.nt® PCV) . Therefont® PCV) . Recentlnt® PCV) . Previount® PCV) . Recentlnt® PCV) . Therefont® PCV) . To datent® PCV) ,298,299.E. coli expression system and the purified capsid protein was capable of self-assembly into virus-like particles (VPLs), which has created hope for the development of VPL-based PCV3 vaccines in the future [E. coli expression system was reported and purified Cap self-assembled into VPLs in vitro with high yield. Such PCV4 VLPs could enter PK-15 and 3D4/21 cells. Furthermore, PCV4 VLP-specific mouse serum showed limited or no cross-reaction with PCV2 and PCV3, indicating significant potential of PCV4 VLPs in PCV4-specific vaccine development [E. coli expression system for the purpose of vaccine preparation for PCV2d and PCV3 was also undertaken, but this attempt did not lead to successful solubilization [At present, no commercial vaccine is available for either PCV3 or PCV4. The protective effect of PCV2 vaccines against PCV3 and PCV4 is also unlikely due to their higher level of genetic and antigenic divergence . Under ee future . Similarelopment . An effolization . Recentllization , as welllization . HoweverAlthough vaccination can prevent clinical disease and economic loss by protecting piglets/sows, the complete eradication of the viruses is not yet possible due to the limited protective period of vaccines and re-infection of viruses has been reported when vaccination was stopped ,305. FurSargassum weizhouense, Epigallocatechin gallate, Astragalus, and saponins were found to inhibit PCV2 replication in both cell cultures and mice [Sophora flavescens Aiton, and scutellarins, a flavonoid from Scutellaria species, were identified as having virucidal activities that interfere with PCV2 replication [Arctium lappa L., has demonstrated a significant inhibitory effect on PCV2 proliferation in PK-15 cells and in mice, which indicates it may be a potential antiviral agent against PCV2 [Spatholobus suberectus Dunn (TFSD) have been shown to reduce the ROS-associated pathologies in RAW264.7 cells, indicating their potential therapeutic use [Due to the lack of effective vaccines, there is an increasing demand for alternative approaches to control multifactorial PCVAD. Many studies have aimed to develop antiviral agents using natural compounds derived from medicinal plants and weeds. Some promising findings have emerged from these efforts. Studies have revealed that certain natural compounds exhibit inhibitory effects on PCV2 replication, both in vitro and in vivo. For instance, the polysaccharides of and mice . In a Chlication . Additionst PCV2 . Furtherutic use . These fImprovement in the management procedures may also help to prevent or lessen PCV2 infection risks and/or the severity of PCVAD. To reduce the severity of clinical diseases in afflicted farms, Madec’s 20-point plan has been proposed . All-in-M. hyopneumoniae 2–4 weeks prior to PCV2 exposure minimizes the severity of PCV2-associated lesions [M. hyopneumoniae alone did not reduce PCV2-induced lesions [M. hyopneumoniae) and trivalent vaccines (containing PCV2a/b and M. hyopneumoniae) under experimental conditions were tested against dual infections (PCV2d and M. hyopneumoniae), which successfully elicited effective protective immunity against concerned pathogens in the form of PCV2-specific neutralizing antibodies and PCV2- and M. hyopneumoniae-specific IFN-γ-SC [M. hyopneumoniae, PCV2, and PRRSV in pigs [As the co-infecting pathogens play a critical role in the clinical outcome of PCVAD, control of such pathogens could also be effective for reducing the severity of disease. It was observed that vaccination against PRRSV significantly reduces clinical signs of PCVAD, as well as the magnitude of PCV2 viremia ,313. How lesions . However lesions . RecentlIFN-γ-SC ,317,318. in pigs . Thus, mOther strategies that can be used to control PCVAD involve the use of disinfectants in buildings and transport vehicles ; the useThis review provides a comprehensive overview of porcine circovirus, covering various aspects such as its structure, replication cycle, interactions with host cells, genetic shifts, epidemiology, and preventive measures through vaccines and management practices. The discussion includes the genomic organization and the role of different proteins in the replication and assembly of various PCV genotypes, including PCV1, PCV2, PCV3, and the recently discovered PCV4. Particular emphasis is placed on PCV2 due to its status as the predominant pathogenic virus worldwide. The review highlights the evolutionary trends of PCV2 genotypes, with PCV2a being the dominant genotype until 2003, followed by the emergence of PCV2b as the prevailing worldwide genotype, and more recently, the predominance of PCV2d since 2012. Detailed insights into the pathogenicity of different PCV genotypes and their ability to evade the host’s immune responses are also discussed. Furthermore, the review addresses the global distribution of PCV, its prevalence in domestic pigs and wild boar, and the factors driving the genotype shifts, particularly the transitions from genotype a to b, and subsequently to d. The article provides a summary of recent developments in preventive strategies against PCV infections, such as inactivated vaccines, subunit vaccines, and virus-like particles (VLPs). The efficacy of these newly developed vaccine candidates is thoroughly compared to that of commercial vaccines. The economic implications associated with PCV’s genotypic shifts for the swine industry are acknowledged, underscoring the need for continued research, surveillance, and the implementation of comprehensive strategies to combat PCV effectively. The review emphasizes the importance of further research and development to enhance our understanding of PCV2 and formulate more effective preventive and control measures.In conclusion, this review highlights the complexity of PCVs and stresses on the different genotypes PCVs to date, and the ongoing efforts required to combat this economically significant pathogen. Through collaborative research and vigilant surveillance, the swine industry can work towards minimizing the impact of PCV and ensuring the health and productivity of pig populations worldwide.