The transcription factor forkhead box (FOXO) controls important biological responses, including proliferation, apoptosis, differentiation, metabolism, and oxidative stress resistance. The transcriptional activity of FOXO is tightly regulated in a variety of cellular processes. FOXO can convert the external stimuli of insulin, growth factors, nutrients, cytokines, and oxidative stress into cell-specific biological responses by regulating the transcriptional activity of target genes. However, how a single transcription factor regulates a large set of target genes in various tissues in response to a variety of external stimuli remains to be clarified. Evidence indicates that FOXO-binding proteins synergistically function to achieve tightly controlled processes. Here, we review the elaborate mechanism of FOXO-binding proteins, focusing on adipogenesis, glucose homeostasis, and other metabolic regulations in order to deepen our understanding and to identify a novel therapeutic target for the prevention and treatment of metabolic disorders. Forkhead box (FOXO) transcription factors play important roles in apoptosis, the cell cycle, DNA damage repair, oxidative stress, cell differentiation, glucose metabolism, and other cellular functions . FOXO geThe transcriptional activity of FOXO is regulated by post-translational modification, including phosphorylation, acetylation, and ubiquitination, which determine subcellular localization, binding with DNA or other regulatory factors, and degradation, among other features. One of the basic regulatory systems is the growth factor-mediated phosphatidylinositol-3 kinase (PI3K)/Akt pathway, which induces phosphorylation of FOXO transcription factors. The three consensus AKT phosphorylation sites are conserved in all members of the mammalian FOXO families, except FOXO6 ,12,13,14FOXO proteins function primarily as transcription factors in the nucleus and bind to the FOXO binding consensus domain of target genes, regulating the expression of these genes. The highly conserved FOXO binding consensus domain, (G/C)(T/A)AA(C/T)AA, is identified as the FOXO-recognized element (FRE) ,15. Furt14-3-3 proteins are a family of modulator proteins that regulate multiple signaling pathways by binding to Ser/Thr-phosphorylated motifs on target proteins ,17,18. 1In contrast, another protein kinase, the AMP-activated protein kinase (AMPK), is essential to the cellular responses to low energy level and activates FOXO function. AMPK has been shown to phosphorylate FOXO3 at six sites  that are different from the AKT phosphorylation sites . In the Oxidative stress also plays an important role in the regulation of FOXO proteins. Under oxidative stress, FOXO translocates to the nucleus to gain stress resistance by activating genes involved in oxidative detoxification, such as manganese superoxide dismutase (MnSOD) and catalase ,27. The The cAMP response element-binding protein (CREB) binding protein (CBP) and its related protein p300 (p300/CBP) are histone acetyltransferases that act as coactivators of numerous transcription factors ,35. In tCaenorhabditis elegans, sir2 and FOXO ortholog daf-16 have overlapping and distinct functions in regulating life span [Silent information regulator 2 (Sir2) is an evolutionarily conserved histone deacetylase in the sirtuin family of nicotinamide adenine dinucleotide (NAD)-dependent deacetylases and is considered as a longevity regulatory gene. Life span extension by Sir2 was first reported in yeast . In Caenife span ,40. Sir2ife span  and induC. elegans [In mammals, the Sir2 family has seven members. Among them, SIRT1 is the closest homologue of yeast and  elegans . SIRT1 i elegans ,43, and  elegans ,45. Acet elegans . A study elegans . Express elegans .KQ/KQFoxo1) and constitutively deacetylated Foxo1 (KR/KRFoxo1) clearly demonstrated the significance of FOXO acetylation in vivo [KQ/KQFoxo1 is predominantly cytoplasmic with a loss of function phenotype, whereas KR/KRFoxo1 is predominantly nuclear with a gain of function phenotype. KQ/KQFoxo1 is embryonic lethal due to cardiac and angiogenic defects. KR/KRFoxo1 mice present with hyperglycemia and insulin resistance. Increased hepatic gluconeogenic gene expression and decreased glycemic gene expression result in excessive hepatic glucose output. Furthermore, KR/KRFoxo1 mice have decreased free fatty acid (FFA) and triglyceride (TG) levels with a lower respiratory quotient, which is consistent with a state of preferential lipid usage. These findings suggest that, in response to fasting or caloric restriction, deacetylated Foxo1 promotes gluconeogenesis, and with prolonged fasting, the gluconeogenesis shifts to lipolysis  to bind to the target region of DNA, and FOXO1 binding to PPARγ is considered to disrupt this PPARγ/RXR complex, resulting in an incapability of PPARγ to bind DNA . PPARγ, Foxo1 is involved in energy homeostasis in adipose tissue ,67,68,69Ucp1 enhancer region and inhibits Ucp1 expression in adipocytes, suppressing energy expenditure. Under cold exposure and other stimuli, Zfp238 binds to Foxo1 and acts as a corepressor, leading to reactivation of Ucp1 expression and the generation of beige adipocytes in WAT  and/or decreased clearance of TG-rich particles is observed. Adipolipoprotein C-III (apoC-III) inhibits lipoprotein lipase and hepatic lipase, impairing the clearance of TG-rich particles and leading to hypertriglyceridemia . The apo insulin . In insueridemia . PPARα wHepatocyte nuclear factor (HNF)-4, a member of the steroid/thyroid nuclear receptor superfamily, is a transcription factor expressed in liver, kidney, intestine, and pancreatic β-cells ,76. HNF-Gck expression: PPARγ, hypoxia-inducible factor-1(HIF-1), sterol regulatory element-binding protein 1c (SREBP1c), and HNF-4 [Gck promoter, which induces Gck expression. Sirt1 deacetylates Foxo1 and deacetylated Foxo1 binds to the Gck promoter and disrupts the binding of HNF-4 to its binding site in the proximal Gck promoter [Gck expression in hepatocyte is tightly controlled not only by Foxo1–HNF-4 interaction but also by the post-translational modification of Foxo1 acetylation (Glucokinase (GCK) is one of the key enzymes that regulates glucose utilization in the liver. Several transcription factors are known to regulate nd HNF-4 . Acetylapromoter . These ftylation .Pgc1 is increased. PGC1α binds and activates FOXO1, leading to activation of gluconeogenic gene expression, such as glucose-6-phosphatase (G6pc) [PGC1α is a transcription co-activator and regulates adaptive thermogenesis in brown fat, muscle-fiber specification, and hepatic-fasting response. In the liver, PGC1α induces gluconeogenesis during fasting . When gle (G6pc) . Foxo1 aPck1), and activates its expression in a C/EBPα-dependent manner. Insulin inhibits Pck1 expression and other C/EBPα-dependent transcription enhanced by Foxo1. Thus, Foxo1 regulates gluconeogenesis by binding to C/EBPα, and by linking insulin signaling to C/EBPα during liver development [CCAAT/enhancer-binding protein (C/EBP) is a family of basic-leucine zipper transcription factors involved in female fertility, gluconeogenesis, adipogenesis, and hematopoiesis . Foxo1 delopment .Gck) expression and increases glucose production by activating G6pc expression [Gck expression in the liver. Ablation of SIN3A in the liver activates the expression of Gck without affecting other FOXO1 target genes, and reduces glycemia without concurrent steatosis. These findings suggest that FOXO1 selectively controls the expression of Gck by interacting with SIN3a. During insulin-induced Gck expression, clearance of SIN3a is observed, which requires phosphorylation of FOXO1 at two sites, Thr24 and Ser253. In contrast, inhibition of G6pc requires FOXO1 phosphorylation of one site, Ser253. These results are consistent with the previous finding that Thr24 phosphorylation is regulated by insulin and insulin-like growth factor-1 receptors [FOXO1 inhibits glucose utilization by suppressing glucokinase  gene, the α-cell determinant, through methylation of the Arx promoter region. In contrast, Foxo1 induces Arx expression by binding to the Arx promoter region. Foxo1 binding to the promoter region releases the DNA methyl transferase (DNMT)3a from the Arx promoter, and hypomethylation in the CG-rich promoter region is induced, leading to activation of Arx expression. In FcorKO, immunostaining shows an increase in Arx-positive endocrine cells from e15.5 and an increase in glucagon-positive cells from e17.5, suggesting that FCoR is involved in α- and β-cell differentiation from the endocrine progenitor stage. These findings suggest that Arx is a target gene of Foxo1 and its transactivation is suppressed by FCoR via methylation by DNMT3a.Furthermore, recent study showed that FCoR is also expressed in pancreatic α- and β-cells from the embryonic stage . FcorKO FcorKO, most Foxo1 translocates to the nucleus and the intrinsic Foxo1 level decreases. This is probably due to deacetylated Foxo1 being activated in the absence of FCoR, followed by degradation. This result is consistent with the previous findings that activated FOXO is rapidly ubiquitinated and degraded [Arx expression and α-cell mass from the endocrine progenitor stage and is required to maintain α- and β-cell identity (FCoR acetylates Foxo1 and suppresses Foxo1 activity in islet cells . The notdegraded . Furtherdegraded . This stidentity .Smooth muscle cells (SMCs) are unique cells with phenotypic plasticity and present transition between a quiescent contractile phenotype and a proliferative phenotype in response to local environmental cues, including growth factors/inhibitors, mechanical influences, cell–cell and cell–matrix interactions, and various inflammatory mediators . Smooth Hes) gene is a Csl target gene. Hes gene expression is activated by Notch/Foxo1 binding, and Hes1 function as a transcription factor that suppresses myogenic determination gene (MyoD) transcription [FOXO transcription regulates myogenesis not only in SMC, but also in skeletal muscle . Conditicription . Thus, NDiabetic cardiomyopathy is defined by the existence of abnormal myocardial structure in the absence of other cardiac risk factors such as hypertension or coronary diseases. In cardiac muscle, insulin receptor substrates (IRS)-1 or IRS-2 activates the Akt signaling pathway and inactivates Foxo1, leading to promoting cardiac function and survival ,100. UndIn contrast, Foxo1 plays an essential role in response to starvation. As mentioned in Pomc and Agrp. Leptin signaling activates Janus kinase 2 (JAK2) and the signal transducer and activator of transcription 3 (STAT3), leading to nuclear translocation of phosphorylated STAT3. STAT3 then activates the Pomc gene while it suppresses the Agrp gene, resulting in suppression of food intake  neurons and orexigenic agouti-related peptide (AgRP) neurons in the arcuate nucleus (ARC) of hypothalamus are the major neuropeptides involved in this process ,103,104.d intake . InsulinSirt1 is expressed in POMC neuron and AgRP neurons in ARC . For hypC. elegans to mammals. Post-translational modification of FOXO tightly controls the activation and/or repression of target gene expression in response to various external stimuli. In addition to the classical regulation by post-translational modifications, there are various regulatory systems involved in FOXO regulation. To achieve this elegant and complex regulatory mechanism, FOXO does not act alone. The partner proteins synergistically function to cope with versatile physiological changes in different organs. Previous studies have shown that there are a variety of patterns regarding interaction between FOXO and the FOXO-binding protein to regulate target gene transcriptions  analysis showed that FOXO3 is related to an extended life span in humans, while FOXO1 is not [The primary function of FOXO is to support the living organism to survive unfavorable conditions and attain longevity. However, in higher eukaryotes, we see that loss-of-insulin signaling, which causes gain-of-FOXO function, can lead to various metabolic disorders, including type 2 diabetes. This is the result of dysregulation of FOXO activity caused by conditions such as obesity and insulin resistance. FOXO function in “longevity” is featured in 1 is not . Loss ofUnderstanding the multilayer mechanism of the tissue-specific metabolic regulation by FOXO and the fine-tuning mechanism by the FOXO-binding protein will lead us to make a proper approach to prevention and treatment of metabolic disorders.


PD-L1 expression than the females, especially in stage III GC. In addition, the patients aged ≥ 65 years had higher PD-L1 expression than the patients younger than 65 years. The multivariate analysis demonstrated that sex was among the independent prognostic factors affecting overall survival (OS) and disease-free survival (DFS). Among the patients with liver metastases, PD-L1 expression was more common among the aged male patients. The males were associated with more tumor recurrence and higher PD-L1 expression than the females, especially in stage III GC. For GC patients with liver metastases, PD-L1 testing is recommended, especially among aged male patients.To date, few reports have investigated the genetic alterations and clinicopathological features in gastric cancer (GC) according to sex. In total, 2673 GC patients receiving curative surgery were enrolled. Among the 2673 GC patients, 1979 (74.0%) patients were male. After propensity-score matching, 846 patients were enrolled for the analysis, including 423 males and 423 females. There was no significant difference in the clinicopathological features between the sexes. Regarding the initial recurrence pattern, the males were more likely to develop tumor recurrence and liver metastasis than the females, especially in stage III GC. Regarding the molecular analysis, the males had higher  Gastric cancer (GC) is the sixth most common cancer and the second most common cause of cancer-related deaths worldwide . SurgicaA male predominance in upper gastrointestinal tract cancers, including esophageal cancer and GC, is observed in almost all populations, and the male gender is a well-established risk factor , 5. SomehMLH3, BRAF, and KRAS between the sexes in young GC patients [PD-L1) expression is reportedly more common in males GC patients [PD-L1 expression between the sexes in GC [PIK3CA amplifications and genetic mutations in the PI3K/AKT pathway, TP53, and ARID1A [It has been reported that in young GC patients, MSI is more common in females than males . Howeverpatients . Programpatients , 9; howees in GC . In addiThe aim of the current study was to compare the clinicopathological features, recurrence patterns, prognoses, and genetic alterations in GC between the sexes after curative surgery.Among the 2673 GC patients, 1979 (74.0%) were males. As shown in As shown in P=0.875). The longest follow-up time was 344.6 months. Among the 846 patients, 207 patients (24.5%) had tumor recurrence during follow-up; 445 patients died during follow-up, and 49 patients (5.8%) had follow-up time shorter than 3 years; among them, there were 22 stage I diseases, 13 stage II diseases, and 14 stage III diseases. Among the 49 patients, four patients had tumor recurrence, including three patients with stage III diseases and one patient with stage II disease.The mean follow-up time was 91.9 months, which was similar between males and females . Patients aged 65 years or older had higher PD-L1 expression than those younger than 65 years .Genetic alterations were analyzed in 438 GC patients with available GC tissues. As shown in P=0.033). In stage III GC, the males were more likely to exhibit PD-L1 expression than the females . Among the 225 stage III GC, the patients aged 65 years or older were more likely to exhibit PD-L1 expression than the patients younger than 65 years .In stage I GC, the females were more likely to have MSI-H tumors than the males  were males, and 12 of the 14 (85.7%) males were aged 65 years or older.Among the 438 patients, forty-five (10.3%) patients, including 39 males and 6 females, developed liver metastases. Among these 45 patients with liver metastases, 16 (35.6%) patients had PD-L1 expression was more common among the aged male patients, especially those with stage III GC. Among the patients with liver metastases, PD-L1 expression was more common among the aged male patients.To minimize selection bias, we performed propensity-score matching and analyzed the clinicopathological feature differences between the sexes in our GC patients. Our results demonstrate that the males are associated with more tumor recurrence, more liver metastasis, and worse prognoses than the females with GC. In addition, the molecular analysis demonstrated that In the present study, sex was one of the independent prognostic factors of OS and DFS. Although gross appearance and pathological T and N categories were significantly related to each other, all of the five independent prognostic factors were still significantly associated with OS and DFS by multivariate analysis. It seems that all the five factors play an important role in GC patient prognosis.P=0.003) in stage I GC only. MSI-H tumors were associated with a better prognosis than MSI-L/S tumors in GC [P=0.436) and DFS  rates were relatively low in both the males and females, resulting in no significant difference in survival rates between the sexes in stage III GC.A meta-analysis  showed trs in GC . The reaPD-L1 expression than the females , and the aged patients were associated with higher PD-L1 expression than the younger patients . In addition, interestingly, among the GC patients with liver metastases and PD-L1 expression, most  patients were males and 12 of the 14 (85.7%) males were aged over 65 years.Among the 438 patients who underwent molecular analysis in the present study, the males had higher PD-L1 expression has been reported to be higher in metastatic liver tumors than primary gastric tumor cells and tumor infiltrating lymphocytes [PD-L1 expression in GC patients who develop liver metastases, especially aged male patients, which may provide useful information for evaluating immunotherapy in these patients.phocytes . ConsequPD-L1 CPS score was developed using the 22C3 assay as a companion diagnostic for immunotherapy in GC, which was approved by the U.S. Food and Drug Administration [PD-L1 expression was more common in male and aged patients, especially in stage III GC, who may be beneficial for adjuvant immunotherapy.The stration . Most trhttp://www.targetscan.org), three miRNAs localized on chromosome X, including miR-106b, miR-20b, and miR-513, are repressors of PD-L1 by direct binding to their 3′UTRs. PD-L1 transcription is induced by hypoxia-inducible factor-1α (HIFα) and signal transducer and activation of transcription-3 (STAT3) factors, directly acting on its promotor [Whole-genome sequencing demonstrated that males had more somatic structural variants than females in GC, but the underlying reason and clinical significance for this result is unclear . Chromospromotor . miR-20bpromotor , thus reIn the study of Sasao et al. , for patRegarding the physiological gastrointestinal motility, for liquid diet, there was no significant difference between sexes; however, for solid diet, the gastric emptying diet is significantly longer in females than males . Pre-menIn our hospital, there has been a case manager in charge of contacting gastric cancer patients for more than 30 years. Consequently, most patients receive regular follow-up at our hospital. Our previous study demonstrated that approximately 90% of gastric cancer recurrences occurred within 3 years after surgery . In the PD-L1 expression in males than females, especially in stage III GC, none of our enrolled patients received immunotherapy for tumor recurrence. We could not evaluate the efficacy of immunotherapy in this group of patients. Further randomized controlled trials investigating the use of immunotherapy for adjuvant therapy are required to validate our results and may provide convincing evidence regarding GC treatment in the future.Because this study is a retrospective study with single center data, this study has limitations. Consequently, we performed propensity-score matching to minimize selection bias. GC, a multifactorial disease, is easily associated with dietary culture and economical levels. Diet with high fat/salt/nitrogen, a history of infection with Helicobacter pylori or Epstein-Barr virus, tobacco use, immune system, and genetic mutations caused by chronic inflammation are risk factors for cancer development –34. In tPD-L1 expression than females, especially in stage III GC. Examining PD-L1 expression is recommended for GC patients with liver metastases, especially aged male patients; immunotherapy may be beneficial for this group of GC patients. Further randomized controlled studies are required to validate our results.In conclusion, males were associated with more tumor recurrence and higher Between January 1992 and December 2013, in total, 2673 GC patients with adenocarcinoma who underwent curative surgery (R0 resection) were enrolled in the present study. Propensity-score matching was performed to minimize selection bias in the comparison of the clinicopathological features between the sexes.th edition of the American Joint Committee on Cancer (AJCC)/Union for International Cancer Control (UICC) TNM classification system [Tumor and normal gastric mucosa tissues were collected and stored in a biobank at our institution. The genetic alterations of 438 GC patients with available tumor and normal gastric mucosa tissues were analyzed. The pathological staging of the GC was performed according to the 8n system .Follow-up examinations, including physical examinations, blood tests with measurements of tumor markers, chest radiography, and sonography or computerized tomography scans of the abdomen, were performed every 3 months during the first 3 years after surgery and every 6 months thereafter.The DNA extraction from the tissue specimens was performed using a QIAamp DNA Tissue Kit and a MinElute Virus Kit  according to a previous report .As described in a previous study , five reTP53, ARID1A, PTEN, PIK3CA, AKT1, AKT2, AKT3, and BRAF) [PI3K/AKT pathway genetic mutations was defined as mutations identified in PIK3CA, PTEN, AKT1, AKT2, or AKT3.A MassARRAY system  was used to identify 68 mutation hotspots in 8 GC-related genes (nd BRAF) . PI3K/AKPD-L1 expression using PD-L1 IHC 22C3 pharmDx kit on a Dako ASL48 platform [PD-L1.IHC staining was performed to detect platform . The comAs shown in 2 test with Yates correction or Fisher’s exact test was used to compare the categorical data. The OS was defined from the date of surgery to the date of death or the last follow-up, while DFS was defined as the length of time after surgery during which the patient was alive without GC recurrence. The Kaplan–Meier method was used for the survival analysis of the OS and DFS. A multivariate analysis with Cox proportional hazards models was performed to analyze the independent prognostic factors of the OS and DFS. A P value < 0.05 was defined as statistically significant.IBM SPSS Statistics 25.0  was used for the statistical analyses. A χ


C9ORF72 was identified as the most common cause of familial amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), three competing but nonexclusive hypotheses to explain how this mutation causes diseases have been proposed and are still under debate. Recent studies in the field have tried to understand how the repeat expansion disrupts cellular physiology, which has suggested interesting convergence of these hypotheses on downstream, functional defects in cells, such as nucleocytoplasmic transport disruption, membrane-less organelle defects, and DNA damage. These studies have not only provided an integrated view of the disease mechanism but also revealed novel cell biology implicated in neurodegeneration. Furthermore, some of the discoveries have given rise to new ideas for therapeutic development. Here, we review the research progress on cellular pathophysiology of C9ORF72-mediated ALS and FTD and its therapeutic implication. We suggest that the repeat expansion drives pathogenesis through a combination of downstream defects, of which some can be therapeutic targets.Ever since a GGGGCC hexanucleotide repeat expansion mutation in  Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are two fatal neurodegenerative diseases with no curative treatment. ALS is a motor neuron degenerative disease with a lifetime risk of one in 347 men and one in 436 women, whereas FTD is the most common form of dementia for people under the age of 45 , 2. ALS Both ALS and FTD are associated with cellular deposition of protein inclusions. For ALS, a pathological hallmark observed in ~98% cases is the mislocalization and aggregation of TAR-DNA binding protein 43 (TDP-43). Other less frequently observed neuropathology includes the aggregation of superoxide dismutase 1 (SOD1), fused in sarcoma (FUS), and some heterogeneous nuclear ribonucleoproteins (hnRNPs) . ForC9ORF72), have been identified to cause both ALS and FTD [Despite their symptomatic differences, ALS and FTD share clinical, neuropathological, and genetic features and are part of a common spectrum. Indeed, ALS and FTD can occur in the same family, and many patients develop signs of both diseases. Furthermore, both ALS and FTD can be related to TDP-43 and FUS neuropathology . In addi and FTD –15.4C2) hexanucleotide repeat expansion (HRE) in C9ORF72 is the most common genetic cause of familial ALS (40%) and FTD (25%) and also presents in some sporadic cases . The lengths of G4C2 HREs are greater than 30 in most patients but vary among individuals, with some patients carrying >1,000 repeats [4C2 HRE causes neurodegeneration is not fully understood. Past studies have suggested that the toxicity arises from one or more of the following assaults  transcript, poly- and  from the antisense (G2C4) transcript, and poly- from both the sense and antisense transcripts.A GGGGCC  lossthe HREs , 17, andthe HREs –22. As t4C2, G2C4 repeat RNA foci, and aggregation of DPRs have been observed in patient tissues and model systems. Furthermore, some of these assaults can indeed cause neurodegeneration and/or are cytotoxic in certain model systems. However, other studies also suggest evidence against any of these three hypotheses. These studies, with a goal of resolving the debate on these three assaults, have been extensively reviewed by others [Consistent with this idea, loss of C9ORF72 mRNA and proteins, Gy others –27.C9ORF72-mediated ALS/FTD (C9ALS/FTD) have related the three assaults to downstream, functional defects in cells. These studies identified molecular and cellular events that are crucial for neurodegeneration implicated in C9ALS/FTD, suggesting novel therapeutic targets for the disease  cells (iPSNs). Using these model systems, research has identified critical cellular events in C9ALS/FTD pathogenesis. We will review the strengths and contributions of different model systems in C9ALS/FTD research.To study the pathomechanism of C9ALS/FTD, many cellular and animal models have been used, including yeast, C. Drosophila do not have a C9ORF72 homolog. However, their short generation time and ease to handle make them powerful genetic tools to study the gain-of-toxicity mechanism. Many yeast or fly models of C9ALS/FTD have been established by ectopically expressing the G4C2 repeat RNA and/or DPRs, which causes cell death or neurodegeneration [C9ORF72 gain of toxicity to arginine-containing DPRs [Drosophila in studying the C9ALS/FTD disease mechanism.Yeast and neration , 28–35. ing DPRs , 33, 34.ing DPRs , 36, 37 ing DPRs , 38–40. C9orf72 is homologous to human C9ORF72 and thus, its knockout (KO) can be used to study the loss-of-function mechanism. However, mouse C9orf72 does not contain G4C2 repeats. Thus, one must ectopically express the repeat RNAs or DPRs in mice, as in yeast and Drosophila, to study the gain-of-toxicity mechanism. Several loss- or gain-of-function mouse models have been established for C9ALS/FTD [Drosophila, and cultured cells [Mouse 9ALS/FTD –46, amoned cells –50 but aed cells .So far, translating drugs identified in animal models to ALS/FTD patients have gained little success –54, suggC. elegans and zebrafish models have also been established to study the C9ALS/FTD mechanism [Besides yeast, fly, mouse and iPSN models, echanism –65. ThesA major challenge in disease research is that all model systems have limitations. Thus, validation across model systems has been a powerful approach in studying human disease pathogenesis. Since non-vertebrate models are quick and easy to handle, whereas mouse and iPSN models are more disease-relevant, an efficient strategy to study disease mechanism is to first use non-vertebrate models to identify potential mechanisms and then, validate the findings in mammals and patient-derived iPSNs. This strategy ensures both the quickness and disease relevance and has been very successful in studying the C9ALS/FTD pathophysiology.C9ORF72 transcript levels are decreased in some C9ALS/FTD patients [C9ORF72 knockdown causes motor neuron degeneration in C. elegans and zebrafish [C9ORF72 KO mice do not exhibit neurodegeneration or motor deficits [C9ORF72 KO enhances the sensitivity of iPSNs to toxic DPRs [Consistent with the loss-of-function hypothesis, patients , 66–70. ebrafish , 65, C9Odeficits , 71–73, xic DPRs , and C9Oxic DPRs . The latxic DPRs , raisingC9ORF72 disrupts endocytosis and autophagy, leading to subcellular aggregation of p62 and/or TDP-43 in cultured neurons [C9ORF72 protein is structurally similar to Rab guanine nucleotide exchange factors (RabGEFs) and functions as a RanGEF when complexing with two other proteins, WDR41 and SMCR8 . RabGEFs neurons . In agre neurons , 80. Sin neurons , these dC9ORF72 KO mice exhibit endolysosomal and/or autophagic defects, but these defects vary across different tissues. In macrophages, C9ORF72 KO impairs autophagy and endolysosomal trafficking [C9ORF72 KO also suppresses mTOR signaling and increases the nuclear level of TFEB, a master regulator of lysosomal biogenesis, in fibroblasts [In agreement with the role of C9ORF72 in cellular vesicle trafficking, fficking , whereasfficking . In addiroblasts , suggestin vitro and in vivo. While insufficient to initiate neurodegeneration, C9ORF72 loss impairs autophagy, which may contribute to neurodegeneration when combined with the gain-of-toxicity mechanism.In summary, C9ORF72 functions as a RabGEF when complexing with other proteins, which regulates vesicle trafficking in cells. Its loss causes endolysosomal and autophagic defects both 4C2 HRE leads to loss of C9ORF72, most evidence suggests that it causes diseases via a gain-of-toxicity mechanism. Indeed, the HRE produces G4C2 and C4G2 repeat mRNAs, which are believed to be responsible for the toxicity. However, how repeat mRNAs cause toxicity is unclear. So far, three models have been proposed: 1) The repeat mRNAs can form guanine-quadruplex (G-quartet) secondary structures, which bind to RNA-binding proteins (RBPs) and lead to their loss of function [While the Gfunction , 17, 37;function ; 3) The function , 83 or sfunction  can undefunction , 85, 86.These three models are nonexclusive, but the first two argue that the repeat mRNAs cause cytotoxicity by their secondary structure, whereas the third one argues that they cause cytotoxicity through DPRs. Consistent with these models, both sense and antisense repeat RNA foci, R-loops, and aggregates of all five DPR species have been observed in C9ALS/FTD patient tissues and model systems , 87, 88.Drosophila models expressing G4C2 repeats interrupted by stop codons in all frames  [4C2 repeats exhibit severe neurodegeneration, suggesting that the G-quartet is non-toxic. This idea is supported by studies in another C9ALS/FTD Drosophila model, which exhibits RNA foci, but no detectable DPR or overt neurodegeneration [Drosophila, whereas in Zebrafish, both sense and antisense “RO” RNAs have been shown to exert neuronal toxicity [Small molecules targeting the RNA secondary structures have been shown to suppress cytotoxicity or neurodegeneration in C9ALS/FTD models, but these molecules also reduce DPR levels , 89, 90.” flies) . These fneration . Howevertoxicity . To bettDespite the controversy over whether the RNA secondary structures play a role in pathogenesis, it is generally agreed that DPRs are cytotoxic. Many studies have shown that DPRs encoded by alternative codons(i.e. non-HRE) cause toxicity in multiple model systems (reviewed in ). In addAmong all five DPR species, the arginine-containing DPRs, i.e. poly(GR) and poly(PR), are the most toxic. In fly models, poly(GR) and poly(PR), but not other DPR species, cause neurodegeneration . ConsistAlthough the repeat RNAs and some DPR species cause little or no toxicity when expressed alone, it is possible that they can enhance poly(GR)- or poly(PR)-mediated cytotoxicity. For example, some RNA has been shown to promote liquid-liquid phase separation (LLPS) of poly(GR) and poly(PR), a process associated with poly(GR)- and poly(PR)-mediated cytotoxicity . FurtherDespite the debate over the RNA-secondary-structure- versus DPR-mediated toxicities, recent studies have suggested that they converge on the same downstream, functional defects in cells, including nucleocytoplasmic transport disruption, membrane-less organelle defects, and DNA damage Figure . Indeed,Several studies have independently identified a critical role of nucleocytoplasmic transport disruption in C9ALS/FTD, which can stem from either RNA secondary structure or DPRs , 37, 102The idea that the G-quartets disrupt nucleocytoplasmic transport by binding to RanGAP1  is furthThe discovery of DPRs disrupting nucleocytoplasmic transport has triggered a surge of further studies, which have identified nucleocytoplasmic transport disruption as a common cellular defect in protein-misfolding diseases –108. UpoStress granules are a group of membrane-less organelles, subcellular compartments that lack surrounding lipid membranes and perform certain biochemical functions. These organelles are protein/RNA condensates enriched in proteins with low-complexity domains (LCDs). Many LCD-containing proteins are aggregate-prone and can undergo LLPS, a process in which molecules demix in an aqueous environment. It has been shown that proper LLPS plays a key role in the assembly, dynamics, and functions of membrane-less organelles, whereas aberrant LLPS can cause aggregation of LCD-containing proteins, as well as defects in these organelles (reviewed in ).in vivo studies have suggested an adverse effect of poly(PR) on heterochromatin protein complexes [Stress GranuleIn C9ALS/FTD, both the sense repeat RNA and DPRs have been shown to disrupt membrane-less organelles, including stress granules and nucleoli. Indeed, poly(GR) and poly(PR) interactomes are enriched in protein components of stress granules, nucleoli, and other membrane-less organelles, and many of these proteins also genetically interact with poly(GR) or poly(PR) , 111. Fuomplexes , anotherStress granules are cytoplasmic RNA/RBP condensates, but certain stimuli can convert them into protein aggregates. Interestingly, several ALS/FTD proteins, including TDP-43, FUS, Ataxin-2, TIA1, and hnRNPs, are stress granule proteins. Since the cytoplasmic aggregation of TDP-43, FUS, and hnRNPs is a pathological hallmark of ALS/FTD, including C9ALS/FTD, stress granules are believed the key to ALS/FTD pathogenesis. So far, this idea has been supported by genetic, cell biology, and pathological evidence in many model systems or patients (reviewed in ). Howeve4C2 repeat RNA and DPRs promote stress granule assembly. The G-quartets formed by the G4C2 repeat RNA bind to and condense RBPs that are stress granule components, promoting RBP LLPS. Consistent with these findings, cells transfected with synthetic G4C2 repeat RNA exhibit stress granules, which occasionally colocalize with the repeat RNAs [in vivo. Consistent with the role of stress granules in translation repression, poly(GR) and poly(PR) have been shown to suppress global translation in multiple model systems [In C9ALS/FTD, both the Geat RNAs . Howevereat RNAs , 118. Imeat RNAs , 101. Aseat RNAs , 49, sug systems , 120.b)NucleolusIn accord with these mechanistic studies, C9ALS/FTD iPSNs exhibit an increased propensity to form, and a decreased rate to clear, stress granules . Importa4C2 repeat RNA and arginine-containing DPRs have been shown to disrupt nucleolar structure and function. Haeusler et al. [4C2 repeat RNA bind to NCL, causing NCL mislocalization and nucleolar stress [Nucleoli are nuclear RNA/RBP condensates where ribosomal biogenesis occurs. They contain many proteins that undergo LLPS, such as NCL and NPM1. Both Gr et al.  have shor stress . Consistr stress  reportedr stress . Howeverr stress , suggestr stress , suggestSeveral studies have shown that either overexpressed or chemically synthesized poly(GR) and poly(PR) localize to nucleoli and disrupt nucleolar functions , 123. Fuin vivo, future studies may focus on whether they disrupt nucleoli when combined with the repeat RNAs and/or other DPRs in mouse models.c)Heterochromatin Protein ComplexDespite their nucleolar toxicity in cultured cells, poly(GR) and poly(PR) rarely localize to nucleoli in C9ALS/FTD patient and mouse models and fail to induce nucleolar stress when overexpressed in mouse brains , 51, 88.In a C9ALS/FTD mouse model overexpressing poly(PR), Zhang et al.  have uned)Other Membrane-less OrganellesHeterochromatin formation is mediated by heterochromatin protein 1 (HP1) proteins, which bind to chromatin and recruit other chromatin-interacting proteins. Recent studies have suggested that the heterochromatin compartment is a membrane-less organelle formed by LLPS of HP1α, an HP1 protein that contains LCDs , 125. InIn addition to stress granule and nucleolar proteins, components of other membrane-less organelles, including nuclear speckles and Cajal bodies, have been identified as physical and/or genetic interactors of poly(GR) and/or poly(PR) , 111. Fu4C2 repeat RNA and DPRs, especially poly(GR) and poly(PR), disrupt multiple types of membrane-less organelles both in vivo and in vitro by binding to and impairing LLPS of proteins in these organelles. Notably, both the G4C2 repeat RNA and arginine-containing DPRs have been shown to undergo LLPS, but whether their LLPS propensity correlates with their toxicity on membrane-less organelles is unclear. Nevertheless, it has been shown that poly(GR) co-aggregates with stress granule proteins in both C9ALS/FTD mouse and patient neurons [In summary, both the G neurons , 109. ThBesides impaired nucleocytoplasmic transport and membrane-less organelles, DNA damage can also be caused by either repeat RNAs or DPRs and has been observed in multiple model systems of C9ALS/FTD, as well as in patients , 127. SoAlthough DNA damage is generally believed to cause neurodegeneration, direct evidence is lacking to demonstrate that it contributes to C9ALS/FTD pathogenesis. Resolving R-loops by Senataxin or suppressing oxidative stress by SOD1 and/or catalase ameliorates repeat-RNA- and/or DPR-mediated toxicity , 126. HoIn addition to nucleocytoplasmic transport disruption, membrane-less organelle defects, and DNA damage, many other functional defects have been implicated in C9ALS/FTD. For example, poly(GR) has been shown to disrupt mitochondrial function by sequestering key mitochondrial proteins in iPSN and/or mouse models , 128. CoEukaryotic cells, including neurons, are highly ordered, dynamic entities with intricate organization. Defects in one cellular organelle or process usually disrupt others. Indeed, many cellular functional defects in C9ALS/FTD are connected. Stress granule assembly has been shown to disrupt nucleocytoplasmic transport in C9ALS/FTD by sequestering transport factors in stress granules , whereasSince the major cause of C9ALS/FTD appears to be the gain of toxicity derived from the repeat RNAs and their translational products, DPRs, methods to eliminate these toxic species have been actively explored. Particularly, an ASO therapy is more advanced than other therapies in clinical development. Notably, the repeat RNAs and DPRs are toxic because they disrupt certain function(s) of cells. Thus, targeting functional defects downstream of the repeat RNAs and/or DPRs holds great therapeutic potential. Indeed, a chemical compound targeting nucleocytoplasmic transport, which stems from our discoveries , is currhttps://clinicaltrials.gov/ct2/show/NCT02623699?term=Tofersen&draw=2&rank=1), supporting the efficacy of ASOs in treating ALS caused by a gain-of-toxicity mutation.ASOs are synthetic single-stranded oligonucleotides that can bind to their target RNAs with very high specificity. This binding can restore, modify, inactivate, or promote the degradation of the RNAs. ASOs are stable and effective in the central nervous system, and an ASO modifying gene expression has been used to successfully treat spinal muscular atrophy, another motor neuron degenerative disease –136. In C9ORF72 have been shown to suppress pathological and pathophysiological defects, as well as neurodegeneration, in multiple model systems [https://clinicaltrials.gov/ct2/show/NCT03626012).For C9ALS/FTD, current ASO therapies focus on degrading the sense transcripts. The ASOs against  systems , 137–139Despite its promise, there are concerns related to ASO therapy. First, the current ASO used in clinical studies does not directly impact the antisense transcript, which may leave part of the pathology unaffected. Furthermore, SMA and ALS1 mainly affect lower motor neurons, whereas C9ALS/FTD is often related to strong upper-motor-neuron affliction. To treat C9ALS/FTD, the pharmacokinetics and pharmacodynamics of ASOs in upper motor neurons must be determined. It is also worth noting that ASOs treating SMA and ALS have different effects on mRNAs—the ASO for SMA blocks an internal splicing site whereas the ASOs for ALS1 and C9ALS/FTD degrade mRNAs. Thus, the clinical effects of these ASOs may be different.4C2 RNA G-quartets or antibodies against poly(GA) have been shown to suppress HRE-mediated toxicity in cell and/or Drosophila models of C9ALS/FTD [https://clinicaltrials.gov/ct2/show/NCT04220021).Small molecules modulating G9ALS/FTD , 90, 140In addition to the ASOs and molecules discussed above, proteins implicated in the production of repeat RNAs or DPRs , 38, 91 https://clinicaltrials.gov/ct2/show/NCT03945279). However, recent studies have suggested that the nuclear export of TDP-43 is not mediated by Exportin-1 [A pathological hallmark of ALS/FTD, including C9ALS/FTD, is cytoplasmic mislocalization of TDP-43 , 9, poteportin-1 , 143 andportin-1 , raisingportin-1 , possiblDrosophila models of C9ALS/FTD [in vivo studies.Stress granule assembly is believed to contribute to ALS/FTD pathogenesis by triggering the aggregation of TDP-43, FUS, and other hnRNPs. Consistent with this hypothesis, inhibiting stress granule assembly using chemical inhibitors or ASOs against Ataxin-2, an essential stress granule component, has been shown to suppress defects in iPSN and/or 9ALS/FTD . Stress 9ALS/FTD . In addi9ALS/FTD , 147. To9ALS/FTD , providiIn addition to nucleocytoplasmic transport and stress granules, other downstream, functional defects may also be therapeutic targets. Importantly, given that some of these defects also present in other types of ALS or FTD, strategies targeting these defects are likely able to be translated from C9ALS/FTD to other ALS/FTD cases. For example, the clinical trial on KPT-350 also includes sporadic ALS patients. However, before targeting a downstream defect, one must carefully evaluate whether this defect is indeed the cause, rather than simply a consequence or byproduct, of neurodegeneration. Thus, rescuing with genetic and pharmacological manipulations in model systems is highly recommended before further analyses.4C2 HRE causes neurodegeneration via a gain of toxicity, emerging evidence suggests that loss of C9ORF72 also plays an auxiliary role. Furthermore, both the loss and gain of function cause defects in many subcellular organelles/processes that contribute to C9ALS/FTD neurodegeneration. Importantly, some of these defects also occur in other forms of ALS/FTD and other neurodegenerative diseases (e.g. Alzheimer’s disease), as well as during cellular responses to a variety of stress. Thus, these studies also help us better understand not only the mechanisms of other diseases but also the principles of fundamental cell biology.Like many other neurodegenerative diseases, C9ALS/FTD has a complex pathophysiology. Although it is widely accepted that the G4C2 HRE occur in parallel or in series. Identifying defects that occur early in pathogenesis is crucial, as targeting these events will likely yield clinical effects.Despite recent progress, several questions remain unanswered. Firstly, some studies suggest the involvement of Golgi and the extracellular matrix in pathogenesis with the underlying mechanism unclear. Secondly, how glia contributes to the diseases is ill-defined. Importantly, it remains unclear whether the downstream defects caused by GIf the ASO approach will treat C9ALS/FTD in the future, as it does for SMA, one may question the value of studying downstream cellular defects. However, many of these defects also occur in sporadic ALS or FTD cases, which lack an obvious genetic cause. Thus, even from a clinical perspective, understanding the cellular and molecular basis of these diseases is still vital.


Chlamydia trachomatis (nvCTs), detected in several countries, cause false-negative or equivocal results using the Aptima Combo 2 assay . We evaluated the clinical sensitivity and specificity, as well as the analytical inclusivity and exclusivity of the updated AC2 for the detection of CT and Neisseria gonorrhoeae (NG) on the automated Panther system (Hologic).Four new variants of We examined 1004 clinical AC2 samples and 225 analytical samples spiked with phenotypically and/or genetically diverse NG and CT strains, and other potentially cross-reacting microbial species. The clinical AC2 samples included CT wild type (WT)-positive (n = 488), all four described AC2 diagnostic-escape nvCTs (n = 170), NG-positive (n = 214), and CT/NG-negative (n = 202) specimens.Trichomonas vaginalis (TV)-positive, bacterial vaginosis-positive, and Candida-positive AC2 specimens gave correct results. The clinical sensitivity and specificity of the updated AC2 for CT detection was 99.7 and 100%, respectively, and for NG detection was 100% for both. Examining spiked samples, the analytical inclusivity and exclusivity were 100%, i.e., in clinically relevant concentrations of spiked microbe.All nvCT-positive samples (100%) and 486 (99.6%) of the CT WT-positive samples were positive in the updated AC2. All NG-positive, CT/NG-negative, Mycoplasma genitalium and TV.The updated AC2, including two CT targets and one NG target, showed a high sensitivity, specificity, inclusivity and exclusivity for the detection of CT WT, nvCTs, and NG. The updated AC2 on the fully automated Panther system offers a simple, rapid, high-throughput, sensitive, and specific diagnosis of CT and NG, which can easily be combined with detection of  Chlamydia trachomatis (CT) infection is the most common bacterial STI with 127 million estimated cases among adults each year globally , spread of the Swedish nvCT escaping detection using the NAATs available from Roche and Abbott at that time –6, 33, aWidespread implementation of validated, accurate and quality-assured CT dual target NAATs for diagnosis of CT is crucial internationally. The updated AC2 assay, now including two CT targets and one NG target , showed


Several derivatives of benzoic acid and semisynthetic alkyl gallates were investigated by an in silico approach to evaluate their potential antiviral activity against SARS-CoV-2 main protease. Molecular docking studies were used to predict their binding affinity and interactions with amino acids residues from the active binding site of SARS-CoV-2 main protease, compared to boceprevir. Deep structural insights and quantum chemical reactivity analysis according to Koopmans’ theorem, as a result of density functional theory (DFT) computations, are reported. Additionally, drug-likeness assessment in terms of Lipinski’s and Weber’s rules for pharmaceutical candidates, is provided. The outcomes of docking and key molecular descriptors and properties were forward analyzed by the statistical approach of principal component analysis (PCA) to identify the degree of their correlation. The obtained results suggest two promising candidates for future drug development to fight against the coronavirus infection. Severe acute respiratory syndrome coronavirus 2 is an international health matter. Previously unheard research efforts to discover specific treatments are in progress worldwide. Virtual screening of existing compound databases against three protein targets  to inhibit coronavirus replication is one of the actual approaches that allows researchers to quickly select best drug candidates for further in vitro assays ,2,3,4,5.Boceprevir is a direct-acting antiviral agent (DAA), acting as an inhibitor of NS3/4A, a serine protease enzyme encoded by hepatitis C virus (HCV) . The serMyrothamnusf labellifolia have proved antiviral activity against herpes simplex virus type-1 by viral adsorption and cell penetration inhibition [Due to their low toxicity and antioxidant activity, phenolic acids and flavonoids appear as the most feasible and secure natural antiviral compounds. The potential antiviral activity of vegetal polyphenols is generally based on their capacity to alter virus replication and functional protein synthesis . Anotherhibition .Furthermore, molecular docking studies in the last two years, made on dozens of natural and synthesized antiviral compounds, associate their antiviral activity with the capacity of their active groups  to bind the active groups of several amino acids found in the active site of SARS-CoV-2 main protease. GLU166, HIS41, ASN142, GLY143, HIS163 and THR 190, are on the top of the most frequent amino acids bound ,18,19,202 from the carboxyl moiety of gallic acid, all other interactions occurred at hydroxyl groups from the flavan ring. Therefore, it is confirmed that the high degree of hydroxylation is associated with a good antiviral effect.Recent virtual screening of 22 U.S.-FDA approved antiviral drugs in the parallel with 24 natural plant-based molecules, indicated theaflavin digallate (from green tea) with the best docking score (−10.574), a result obtained using the Glide (Schrödinger) module on the COVID-19 main protease (structure 6LU7) . It mustAnother aspect to be mentioned is the conclusion of a computational survey to a drug repurposing study claiming that five neutral antiviral drugs have inhibitory activities against SARS-CoV-2 main protease . As it iGallic acid, one of the most abundant phenolic acids in plants, has various health benefits including anti-inflammatory, antioxidant and antimicrobial activities  and is vStarting from such intriguing premises, in molecular docking approach we conducted in silico screening on several benzoic acid derivatives and on a homologue series of alkyl gallates, starting from the lead compound gallic acid, against SARS-CoV-2 main protease.We selected the following compounds: benzoic acid, 4-aminobenzoic acid, 4-hydroxybenzoic acid, 3,4,5-trimethoxybenzoic acid (eudesmic acid), 3,4-dihydroxybenzoic acid (protocatehuic acid), 2,5-dihydroxybenzoic acid (gentisic acid), 4-hydroxy-3-methoxybenzaldehyde (vanillin), 4-hydroxy-3,5-dimethoxybenzoic acid (syringic acid), 4,5-dihydroxy-3-oxocyclohex-1-ene-1-carboxylic acid, epicatechin, 3,4,5-trihydroxybenzoic acid , methyl 3,4,5-trihydroxybenzoate, ethyl 3,4,5-trihydroxybenzoate, propyl 3,4,5-trihydroxybenzoate, isopropyl 3,4,5-trihydroxybenzoate, butyl 3,4,5-trihydroxybenzoate, isobutyl 3,4,5-trihydroxybenzoate, pentyl 3,4,5-trihydroxybenzoate, isopentyl 3,4,5-trihydroxybenzoate, octyl 3,4,5-trihydroxybenzoate. Their 3D optimized structures obtained with the Spartan program, along with the atomic numbering scheme, arbitrary chosen by the software, are given in Intermolecular interactions occurring in boceprevir, benzoic acid and alkyl gallate derivatives in complex with the 6WNP protein fragment were identified. The lengths of hydrogen-bonding interactions were measured. The results are given in terms of docking score function and RMSD (root mean square deviation).To validate the molecular docking protocol, boceprevir was initially docked into the crystal structure of the main protease fragment and its interactions with the target 3C-like proteinase were analyzed. As can be seen 2 CYS145 (2.900 Å), N sp2 SER144 (3.053 Å), N sp2 GLY143 (2.783 Å), N sp2 HIS41 (2.604 Å), O sp2 HIS164 (3.103 Å), N sp2 GLU166 (3.118 Å), O sp2 GLU166 (2.908 Å and O sp2 GLU166 (3.229 Å). Remarkable is the docking score for octyl-gallate , close to boceprevir, and for epicatechin (−49.57). Benzoic acid derivatives, in the first half of the graph exhibit moderate scores ranging between −29.59 (benzoic acid) and −37.25 (syringic acid). We observed an increasing trend of score with the increasing number of hydroxyl groups. For instance, gallic acid docking simulations resulted in a −38.31 score. Regarding the gallates, the increasing length of alkyl chain led to docking scores in the order gallic acid < methyl gallate < ethyl gallate < isopropyl gallate < propyl gallate < butyl gallate < isobutyl gallate < isopentyl gallate < pentyl gallate < octyl gallate. The ramification of lateral side chain led to a slight decrease of binding affinity, noticeable with an increasing number of -CH2 .In 2 GLU166, N sp2 HIS163, O sp3 SER144, O sp3 SER144, O sp2 LEU141, N sp2 SER144, N sp2 GLY143, N sp2 CYS145, N sp2 GLY143 and O sp2 ASN142. Their obtained docking scores were −42.13 and −48.77, respectively; lower than for octyl-gallate but greater than the results for gallic acid (−38.31). These observations are in good agreement with experimental findings of Takai E. et al., 2019 [n-alkyl side chain. The good docking result for octyl gallate recommends it as good alternative for developing new therapeutic antiviral agents.Amino acid residues CYS145 and SER144 interacted by hydrogen-bonding both with boceprevir and with octyl-gallate see a; similal., 2019 , anticipl., 2019 ,26,27. T2 and with a maximum 10 rotatable bonds for good oral bioavailability.In −1) and six hydrogen bond donors. The structure of boceprevir presents the maximum allowed number of flexible bonds (10) and maximum hydrogen bond acceptors (10). Although it had these exceptions, the docking score was the highest among the investigated ligands, suggesting strong interactions and stability of the complex formed with the SARS-CoV-2 main protease. The results indicated that all proposed ligands met the restrictive criteria for good oral bioavailability. Increased hydrophilicity was observed for all compounds due to their hydroxyl, carboxyl and/or methoxy groups on their skeleton. These functional groups offer good absorption and permeation properties. Thus, by means of NH/OH/N/O groups, hydrophilic interactions were favored and further reflected in good and high docking scores. Concerning logP values, there were observed positive values for benzoic acid (0.79), 2,5-dihydroxybenzoic acid (0.81) and 0.72 for octyl 3,4,5-trihydroxy benzoate. A combination of molecular factors and properties, mainly due to the increased hydrophobicity of the lateral n-octyl chain of octyl gallate, was also reflected by its best docking score function, indicating this compound as the best antiviral candidate among all screened compounds in the study.Therefore, the boceprevir antiviral exhibited two violations of Lipinski’s criteria, namely molecular mass  and the lowest unoccupied molecular orbital (LUMO) localization and energy levels for octyl-gallate, are illustrated in HOMO) and electron affinity (A = −ELUMO), according Koopmans’ theorem [The resulting band gap (ΔE) can provide useful information on the chemical reactivity and kinetic stability of each ligand. The same values for energy gap were given for alkyl gallates, starting at ethyl to octyl-gallate, suggesting the same stability. Slight differences were found for the values of ionization potential , refers to the energy needed to remove an electron from a molecule, and electron affinity (A) measures the ability of a molecule to accept electrons and form anions species ,35. SuchPrincipal component analysis (PCA) is a statistical tool for the identification of linear combinations of the variables which account for certain proportions of the variance of a set of variables. The selection is based on the eigenvalues of the dispersion matrix of the variables. The principal components are associated with decreasing eigenvalues and, therefore, share the amount of the variance. Typically, the first few principal components account for virtually all the variance. PCA also represents the pattern of similarity of the observations and the variables by displaying them as points in maps ,37,38,39PCA analysis was employed to find the degree of correlation between molecular descriptors and properties and their involvement in the resulting docking score.The PCA correlation matrix  revealedn = 6) to a lower number of dimensions.From 3, 40.45 Å3, 36.62 Å3. Redocking of the cocrystallized ligand (boceprevir) was realized to validate the docking protocol. The amino acid residues forming the binding site were protonated and water molecules were removed.The docking simulation was carried out using CLC Drug, Discovery Work Bench . SARS-CoV-2 main protease bound to boceprevir at 1.45 Å (PDB ID: 6WNP) , which whttps://pubchem.ncbi.nlm.nih.gov) or directly constructed  in the Spartan’18 program [The examined ligands were first generated in 3D by importing their corresponding files from the Pubchem database , Copyright (2013–2020) Charles Zaiontz .The predicted ligand data were processed to compute principal component analysis using the free Excel add-in Real Statistics Resource Pack software  suggested 2,5-dihydroxybenzoic acid (gentisic acid) and octyl gallate as the best potential candidates among the investigated structures. The two compounds had similar logP values (0.81 and 0.72). Octyl gallate had the best docking score (−60.22), but decreased dipole moment (1.31). 2,5-dihydroxybenzoic acid (gentisic acid) had a lower score (−33.84) but an increased dipole moment (4.62) which means higher polarity and also higher reactivity. As is known, the dipole moment represents a measure of net molecular polarity and, therefore, the larger the difference in electronegativities of bonded atoms the larger the reactivity of the molecule. Accordingly, a combination of the two compounds can be considered. On the other hand, if the benzoic acid series indicated that the position of hydroxyl groups on the ring was more important than the ester, hydroxyl and methoxy groups’ number or type, the galllic acid series clearly indicated similar results for all studied compounds, apart from octyl gallate which was proved to be the best potential candidate among investigated structures, exhibiting antiviral activity against the SARS-CoV-2 protease and, therefore, could be studied further for drug development.


NLRP3 gene) is expressed predominantly in macrophages as a component of the inflammasome and is associated with many diseases, including gout, type 2 diabetes, multiple sclerosis, atherosclerosis, and neurological diseases and disorders. Diterpenes containing repeated isoprenoid units in their structure are a member of some essential oils that possess diverse biological activities and are becoming a landmark in the field of drug discovery and development. This review sketches a current scenario of diterpenes or their derivatives acting through NLRPs, especially NLRP3-associated pathways with anti-inflammatory effects. For this, a literature survey on the subject has been undertaken using a number of known databases with specific keywords. Findings from the aforementioned databases suggest that diterpenes and their derivatives can exert anti-inflammatory effects via NLRPs-related pathways. Andrographolide, triptolide, kaurenoic acid, carnosic acid, oridonin, teuvincenone F, and some derivatives of tanshinone IIA and phorbol have been found to act through NLRP3 inflammasome pathways. In conclusion, diterpenes and their derivatives could be one of the promising compounds for the treatment of NLRP3-mediated inflammatory diseases and disorders.Nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing protein (NLRP) inflammasomes are involved in the molecular pathogenesis of many diseases and disorders. Among NLRPs, the NLRP3 (in humans encoded by the  Inflammation, the body's natural response to harmful stimuli, arises in tissues due to traumatic, infectious, post-ischemic, toxic, or autoimmune injuries. To restore the normal tissue functions during the harmful inflammatory conditions, clearance of inflammatory cells, along with pro-inflammatory signaling pathways, are required . The nucNon-steroidal anti-inflammatory drugs (NSAIDs) are the most used medications for the treatment of inflammation and related diseases. However, NSAIDs cause many adverse effects, including gastrointestinal complications, immunodeficiency, and humoral disturbances, which may limit their use in certain patients . The natRecently, Tricarico and coworkers suggested that isoprenoids may exert their effect by modulating different pathways in the biological systems primarily by increasing the expression of various markers for apoptosis, cytokines, mitochondrial dysfunction, NLRP3, as well as nitric oxide (NO) . Based oClinicalTrials.gov, using the keywords “diterpenes or diterpenoids” and “diterpene derivatives,” combined with “NLRP3 or NLRP3 inflammasome.” No language restrictions were imposed. The following are the inclusion and exclusion criteria of this study.A literature search was conducted through June 2020 by means of a number of well-known databases, such as Google Scholar, PubMed, Scopus, ScienceDirect, Web of Science, the American Chemical Society, and Studies with diterpenes and their derivatives or preparations acting against NLRP3;in vitro, ex vivo, or in vivo) with or without using experimental animals, including humans and their derived tissue and cells;Studies ;Studies with or without proposing activity mechanisms.Studies with extracts without mentioning diterpene contents;Studies with essential oils other than diterpenes;Duplication of data and titles and/or abstracts not meeting the inclusion criteria;Diterpenes acting against NLRPs other than NLRP3;Diterpenes with other studies uncovering the current topic.Macrophages stimulated by priming stimuli, such as ligands for toll-like receptors (TLRs), NOD-like receptors (NLRs) , or cytokine receptors, can activate nuclear factor kappa B (NF-κB). The signaling molecules MyD88 and TRIF of the NF-κB signaling pathway regulate the induction of NLRP3 and pro-interleukin 1 beta (IL-1β) in response to TLR ligands . HoweverReactive oxygen species (ROS) are proposed as a common signal for NLRP3 inflammasome . MitochoResearch findings revealed that phytanol (Compound 1) and phytanyl amine (Compound 2) reduce the expression of NLRP3, along with interleukin (IL)-6, monocyte chemoattractant protein-1 (MCP-1), keratinocyte chemoattractant (KC), macrophage inflammatory protein 1 (MIP-1), and lipopolysaccharide-induced CXC chemokine  . Additiovia the attenuation of transverse aortic constriction-induced myocardial remodeling. This effect (a) enhanced cardiac diastolic and systolic functions, (b) activated the pro-fibrotic tumor growth factor 1 beta (TGF-1β) pathway, (c) suppressed the NLRP3 inflammasome and inflammatory mediators of IL-1β, IL-18, MCP-1, and vascular cell adhesion molecule-1 (VCAM-1), and (d) dose-dependently inhibited macrophage infiltration (via targeting the NLRP3 inflammasome (Triptolide (Compound 3) significantly reduced the serum levels of IL-1β and IL-18, along with NLRP3 and toll-like receptor 4 (TLR4), expressions in female Sprague–Dawley rats . It may ltration . In a reammasome . Furtherammasome .n = 15) treated with this compound at 10 mg/kg/day (i.p.) for 1 week down-regulated tumor necrosis factor alpha (TNF-α), IL-4, TLR4, MyD88, NLRP3, and NF-κB p65 expression levels and up-regulated IL-10, tumor growth factor-beta (TGF-β), phosphatase and tensin homolog (PTEN), phosphoinositide 3-kinase (PI3K), and AKT  phosphorylation levels /p38 mitogen-activated protein kinase (MAPK) signaling pathway  (Tanshinone IIA (Compound 5) at 0–2.5 μg/ml exerted a protective effect by down-regulating NLRP3, caspase-1 (CASP1), IL-1β, and IL-18 in BV-2 cells . The maln levels . Liu and pathway . On the  pathway . In anotB/c mice . PhorbolB/c mice . In conte (Daoy) .via the PIK3CA–AKT1–MTOR–RPS6KB1 pathway. It additionally increased the disruption of NLRP3–PYCARD–CASP1 complex formation and mitophagy in macrophages, inactivated the NLRP3 inflammasome, and induced autophagy in mice (l-amino acid defined (CDAA)-fed mice; this was accomplished by reducing the hepatic inflammation and fibrosis. It also lowered the hepatic mRNA levels of both pro-inflammatory and pro-fibrotic genes, as well as hepatic macrophage infiltration, and reduced the expression of inflammasome genes, IL-1β expression through NF-κB inhibitory pathway, and inflammasome disassembly (Andrographolide (Compound 10), the bitter diterpene lactone, at 7.5 and 15 mg/kg exerted an anti-colitis and anti-tumor effect by lowering the expression of cleaved CASP1, IL-1β, and mitochondrial membrane potential collapse  in mice . In anot in mice . Furtherassembly . In addiassembly .Carnosic acid (Compound 11) lowered the expression of NLRP3 and CASP1, along with myeloperoxidase (MPO) levels, in male BALB/c mice. Additionally, it increased the level of nuclear factor erythroid 2 (Nrf2) and stopped the degradation of Nrf2 through a mechanism that involves impeding the interaction between Cullin3 and Keap1. It also increased the reduced glutathione (GSH) and superoxide dismutase (SOD) levels and caused a decrease in malondialdehyde (MDA) and iNOS levels in experimental animals . In wild20H32, which is composed of repeated isoprene subunits. Diterpenes and their derivatives are known for their antimicrobial and anti-inflammatory activities  and damage-associated molecular patterns (DAMPs) that trigger its activation . The higtivities . In thistivities .For the activation of pro-CASP1, research findings showed that caspase activation and recruitment domains (CARDs) act as a platform termed the ASC speck . Along tNACHT, LRR, and PYD domains-containing protein 12 (NLRP12), a member of the NLRs, which is known to interact with the apoptosis-linked speck-like protein containing a CARD (ASC), cause the activation of CASP1, resulting in IL-1β release . RecentlNLRP3 gene occurs, which activates NF-κB, including TLRs and TNF receptors  are known to regulate lipid homeostasis and activate the transcription of genes encoding enzymes involved in the biosynthesis of compounds including cholesterol, triglycerides, phospholipids, and fatty acids. In this regard, SREBP-1c, one of the three SREBP isoforms, activates genes that are involved in fatty acid synthesis ; this isThe opening of the P2X7 channel may lead to the accumulation of high levels of extracellular ATP  when thevia ubiquitination by blocking the interaction between Cullin3 and Keap1  plays a regulatory role in the NLRP3 inflammasome. Nrf2 constantly degrades upon ubiquitination under normal conditions. These inflammasomes represent stress-induced protein complexes that are involved in acute and chronic inflammation by means of CASP1-mediated activation of pro-inflammatory cytokines . Carnosind Keap1 . In addind Keap1 . Moreovend Keap1 . TXNIP tnd Keap1 . Sodium  pathway . In this release . The voltivation . Tanshintivation , androgrtivation , and cartivation  were seetivation , 84. Tantivation , whereastivation  in expervia interaction with the NLRP3 inflammasome.In primary human macrophages, p38δ MAPK has been recognized as a regulator of NLRP3 inflammasome activation, which might be one of the potential targets for the treatment of atherosclerotic inflammation . On the A) receptor. GABA, the major inhibitory neurotransmitter in the brain, after binding to benzodiazepines, increases the total conduction of chloride ions across the neuronal cell membrane and causes chloride ion influx, hyperpolarizing the neuron's membrane potential. Therefore, the difference between resting potential and threshold potential is increased, and firing is less likely. As a result, arousal of the cortical and limbic systems in the central nervous system is reduced  are positive allosteric modulators of the neurotransmitter gamma-aminobutyric acid (GABA reduced . To exer reduced . In fact reduced .The recent human threat SARS-CoV-2 is known to produce cytokine storm in Covid-19 patients. It is evident to release pro-inflammatory cytokines, including IL-6 and IL-1β through the NLRP3 inflammasome pathway . Many navia the activation of CASP1. The NLRP3 inflammasome is able to act through the mitochondria; therefore, it has an important impact on physiological homeostasis and metabolic balance. Furthermore, this review has shed some light on 13 important diterpenes that act through the NLRP3 inflammasome interactive pathway, providing new insights for the development of new therapeutic modalities.At the present time, the utilization of natural products and their derivatives has gained popularity due to the lesser side effects and economy as compared with other treatment strategies. Cumulative research papers have dealt with the health-promoting benefits of diterpenes and their derivatives as they could be used as chemotherapeutic tools against many diseases, including cancer, diabetes, obesity, and neurological diseases and disorders. Findings from this review highlight the role of diterpenes and their interaction with the NLRP3 inflammasome. In addition, this review has shown that NLRP3 inflammasome can be an important molecular platform in the induction of the central pro-inflammatory cytokine IL-1β MI and JS-R: conceptualization. SB: validation investigation. MI, SB, MM, WK, KG-B, BA, and JS-R: resources. MI, SB, and MM: data curation. MI, JS-R, and WK: review and editing. All the authors writing, read, and approved the final manuscript. All authors contributed equally to the manuscript.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.


The primary objective is to determine which of two interventions: 1) an eight week, online, home-based, supervised, group rehabilitation programme (REGAIN); or 2) a single online session of advice ; is the most clinically and cost-effective treatment for people with ongoing COVID-19 sequelae more than three months after hospital discharge.Multi-centre, 2-arm (1:1 ratio) parallel group, randomised controlled trial with embedded process evaluation and health economic evaluation.Adults with ongoing COVID-19 sequelae more than three months after hospital dischargeInclusion criteria: 1) Adults ≥18 years; 2) ≥ 3 months after any hospital discharge related to COVID-19 infection, regardless of need for critical care or ventilatory support; 3) substantial (as defined by the participant) COVID-19 related physical and/or mental health problems; 4) access to, and able/supported to use email and internet audio/video; 4) able to provide informed consent; 5) able to understand spoken and written English, Bengali, Gujarati, Urdu, Punjabi or Mandarin, themselves or supported by family/friends.Exclusion criteria: 1) exercise contraindicated; 2) severe mental health problems preventing engagement; 3) previous randomisation in the present study; 4) already engaged in, or planning to engage in an alternative NHS rehabilitation programme in the next 12 weeks; 5) a member of the same household previously randomised in the present study.Intervention 1: The Rehabilitation Exercise and psycholoGical support After covid-19 InfectioN (REGAIN) programme: an eight week, online, home-based, supervised, group rehabilitation programme.Intervention 2: A thirty-minute, on-line, one-to-one consultation with a REGAIN practitioner .The primary outcome is health-related quality of life (HRQoL) – PROMIS® 29+2 Profile v2.1 (PROPr) – measured at three months post-randomisation.Secondary outcomes include dyspnoea, cognitive function, health utility, physical activity participation, post-traumatic stress disorder (PTSD) symptom severity, depressive and anxiety symptoms, work status, health and social care resource use, death - measured at three, six and 12 months post-randomisation.Participants will be randomised to best practice usual care or the REGAIN programme on a 1:1.03 basis using a computer-generated randomisation sequence, performed by minimisation and stratified by age, level of hospital care, and case level mental health symptomatology.Once consent and baseline questionnaires have been completed by the participant online at home, randomisation will be performed automatically by a bespoke web-based system.To ensure allocation concealment from both participant and REGAIN practitioner at baseline, randomisation will be performed only after the baseline questionnaires have been completed online at home by the participant. After randomisation has been performed, participants and REGAIN practitioners cannot be blind to group allocation. Follow-up outcome assessments will be completed by participants online at home.A total of 535 participants will be randomised: 263 to the best-practice usual care arm, and 272 participants to the REGAIN programme arm.Current protocol: Version 3.0 (27th October 2020) Recruitment will begin in December 2020 and is anticipated to complete by September 2021.ISRCTN:11466448, 23rd November 2020The full protocol Version 3.0 (27th October 2020) is attached as an additional file, accessible from the Trials website  guidelines. Additional file 1.


Improving the therapeutic efficacy of conventional anticancer drugs represents the best hope for cancer treatment. However, the shortage of druggable targets and the increasing development of anticancer drug resistance remain significant problems. Recently, membrane transport proteins have emerged as novel therapeutic targets for cancer treatment. These proteins are essential for a plethora of cell functions ranging from cell homeostasis to clinical drug toxicity. Furthermore, their association with carcinogenesis and chemoresistance has opened new vistas for pharmacology-based cancer research. This review provides a comprehensive update of our current knowledge on the functional expression profile of membrane transport proteins in cancer and chemoresistant tumours that may form the basis for new cancer treatment strategies. In mammalian cells, the plasma membrane is a selectively permeable barrier that creates an intracellular environment and maintains cell stability and homeostasis. The proper functioning of the plasma membrane is dependent on a group of membrane transport proteins that permit the selective transport of essential substances for the survival and development of the organism . To dateMembrane pumps are transmembrane proteins that facilitate the active transport of various substances against their electrochemical gradients. Mechanistically, membrane pumps can be divided into two main categories: primary and secondary active transporters. Through ATP hydrolysis, primary active transporters move solutes against their electrochemical gradients. These pumps are often uniporters which are involved in the active transport of a single molecule across the cell membrane. Instead, secondary active transporters utilize the energy stored in the electrochemical gradient of ions across the plasma membrane that was generated by the primary active transporters. Therefore, in this type of transport, the transfer of one molecule down its gradient is coupled to the movement of another molecule against its gradient A. DependThe crucial role of membrane pumps in conducting the active transport of a wide range of substrates including ions, amino acids, large polypeptides, and essential metabolites highlights their indispensable function in maintaining cellular homeostasis . Moreove+/K+-ATPase) is a hetero-dimeric complex that consists of catalytic a- and regulatory b-subunits , respectively [+/K+-ATPase and etiology of several malignancies, including breast, non-small cell lung cancer, glioblastoma, and melanoma has been established [+/K+-ATPase is highly expressed in glioblastomas and that its inhibition decreases cell proliferation and migration while increasing survival of the orthotopic patient-derived xenograft mouse model of human glioblastoma [+/K+-ATPase positively impacts E-cadherin-mediated formation of tight junctions and epithelial cell polarity [1-subunit correlates with poorly differentiated breast (MDA435), colon (SW480), pancreas (MiaPaCa-2), and kidney (MSV-MDCK) cancer cells. This evidence suggests that the Na+/K+-ATPase β1-subunit plays a tumour-suppressor role in cancer [The plasma membrane sodium pump  is a P-type ATPase located in the sarcoplasmic reticulum (SR) within myocytes [2+ binding and transport  is a large multi-subunit P-type ATPase, present in both vacuolar membranes and plasma membranes and which is involved in controlling cellular pH . Structumembrane ,82. The membrane ,84. Heremembrane ,86,87.In tumours, the expression level of V-ATPase is often upregulated . For insFurthermore, overexpression of V-ATPase has been reported to be closely associated with the development of chemoresistance ,103. ForIon channels are gated aqueous pores involved in the selective movement of ions across biological membranes. In this type of transport, ions are passively moved down their electrochemical gradient . Dependi2+ is a universal second messenger that is vital for the proper functioning of the organism [2+ channels, voltage-gated Ca2+ channels, Orai-mediated store-operated channels, and transient receptor potential-mediated Ca2+ channels. It has been revealed that disruption in the expression or function of these channels is often correlated with carcinogenesis and/or chemoresistance [2+ channels in cancer progression and chemoresistance.Caorganism . Thus, isistance ,120. HenV) are Ca2+ channels that open in response to membrane depolarization. Each CaV consists of a central a1 subunit and three auxiliary subunits, a2δ, b, and g, in a 1:1:1:1 ratio. In mammals, ten distinct members are grouped into three phylogenetic subfamilies: CaV1 , CaV2 , and CaV3  [V channels are functionally expressed in non-excitable cancer cells [V subfamilies have been observed in various cancers, suggesting their role in tumour progression, differentiation, and invasion [V1.2 encoded by the human CACNA1C gene is predominantly expressed in oesophageal squamous cell carcinoma and is correlated with tumour cell differentiation [V1.3 encoded by the human CACNA1D gene is overexpressed in prostate and endometrial cancers [V1 inhibitor BK10040 was reported to reduce proliferation and induce apoptosis in cancer cells such as A459 (lung adenocarcinoma) and MiaPaCa2 (pancreatic cancer cells) cell lines. Together, these pieces of evidence reveal the oncogenic role of the CaV1 subfamily in human cancers.VGCCs (CaaV3.1–3) . Historier cells . Interesinvasion ,124,125.ntiation . Likewis cancers ,128. HerV1 subfamily, different CaV2 channels have been reported to be dysregulated in cancer. For example, CaV2.3 encoded by the human CACNA1E gene is upregulated in Wilm’s tumours (a rare childhood kidney cancer), and its expression level is associated with reduced relapse-free survival [V3.1 and CaV3.2 are highly expressed in human laryngeal carcinoma and glioblastoma, respectively, and their inhibition (using siRNA or mibefradil) causes cell cycle arrest and apoptosis [V channels has been also shown to be associated with chemoresistance. For example, overexpression of the regulatory subunit α2δ (encoded by human CACN12D3 gene) sensitized esophageal squamous cell carcinoma cell lines to cisplatin-induced cell death. Therefore, genetic silencing of α2δ promoted cisplatin resistance in vitro and in vivo [V3 subfamily) and carboplatin synergistically inhibited the growth of platinum-resistant ovarian cancer cell lines A2780Cis and IGROV-1, suggesting that a combinatorial drug therapy using CaV inhibitors and conventional chemotherapeutics may serve as an effective treatment for ovarian cancer [As members of the Casurvival . Similarpoptosis ,131. Mor in vivo . Likewisn cancer .2+ channels that open in response to reduced ER Ca2+ levels. Hence, these channels are called store-operated channels (SOC). Currently, three Orai isoforms have been described , and each of them consists of six subunits that form a single pore [Orai proteins are highly selective Cagle pore . In cancgle pore ,136,137.gle pore . Similargle pore . Like Orgle pore ,141. SimFurthermore, the expression profile of Orai channels has been associated with chemoresistance in many cancers . For exa2+, either directly by conducting Ca2+ entry or indirectly by changing membrane potential and providing a driving force for Ca2+ entry by other channels [The transient receptor potential (TRP) family of ion channels consists of 28 distinct members divided into seven subfamilies: TRPC , TRPV (Vanilloid), TRPM (Melastatin), TRPML (Mucolipin), TRPP (Polycystin), TRPA (Ankyrin), and TRPN  . Structuchannels ,151. Thechannels ,155,156.2+-dependent cellular processes [2+ entry and reduces cell proliferation [The TRPC subfamily includes seven members (TRPC1–7) which are ubiquitously expressed and play important roles in the regulation of several Carocesses . In cancrocesses , therefoferation ,160. Simferation . Moreoveferation . Furtherferation . Similarferation , gastricferation , and breferation , wherebyferation . Therefoferation .Moreover, the altered expression profile of various members of the TRPC subfamily has been associated with chemoresistance. For instance, TRPC1 expression is significantly decreased in cisplatin-resistant (A2780 and SKOV3) and carboplatin-resistant (A2780) ovarian cancer cell lines, suggesting that the reduced expression of TRPC1 is linked to chemoresistance . On the 2+ permeability, ranging from Ca2+- impermeable channels  to highly Ca2+-permeable channels (TRPM6/7) [The TRPM subfamily consists of eight members, TRPM1–8, and each member represents different CaTRPM6/7) . AlteredTRPM6/7) ,176,177.TRPM6/7) ,179. FurTRPM6/7) ,181,182.TRPM6/7) . Like TRTRPM6/7) . For exaTRPM6/7) ,186, breTRPM6/7) , ovarianTRPM6/7) , and blaTRPM6/7)  cancers TRPM6/7) ,189, panTRPM6/7) ,191, andTRPM6/7) ,193 cancTRPM6/7) .Furthermore, the altered expression of several TRPM channels has been associated with anticancer drug resistance. For instance, inhibition of TRPM2 expression or function increased the cytotoxic effect of paclitaxel and doxorubicin in breast and gastric cancer cells ,195. A s2+. While TRPV1–V4 are modestly permeable to Ca2+, TRPV5 and TRPV6 represent high Ca2+ selectivity. In cancer, the expression profile of TRPV channels is highly contextualized; therefore, depending on the cell and tumour type, TRPV channels can act as both tumour promoters and suppressors [All six members of the TRPV subfamily show variable permeability to Capressors . For inspressors . Similarpressors ,202,203.pressors . On the pressors ,206,207.Furthermore, enhanced expression of TRPV2 in triple-negative breast cancer , bladderIn addition to the biological roles and prognostic values, TRPVs have been reported to be involved in the regulation of chemoresistance. In this regard, activation of the TRPV1 channel has been shown to enhance the cytotoxic effects of 5-FU , cisplat+ channels) are a diverse and ubiquitous group of ion channels involved in the maintenance and regulation of K+ gradients. Given the essential role of K+ in the control of cell homeostasis and functions, the proper functioning of K+ channels is crucial for a wide array of cellular functions, ranging from membrane excitability to cell proliferation, migration, and apoptosis [+ channels have been identified and divided into four main classes based on their structural and biophysical characteristics: voltage-gated K+ channels (Kv) [2+- activated K+ channels (KCa) [+ channels (Kir) [+ channels (K2P) [Potassium channels (Kpoptosis . Currentels (Kv) , Ca2+- als (KCa) , inwardlls (Kir) , and twols (K2P) . Kv, KCals (K2P) . On the ls (K2P) .+ channels have been associated with the etiology of many cancers [+ channels show differential expression patterns between different cancers and within the same cancer. For instance, Kv11.1 is overexpressed in HT-29 colorectal cancer cells while it was found downregulated in lung carcinoma A549 cells [Alterations in the functional expression of K cancers ,241. For cancers . Similar cancers , breast  cancers , and cle cancers , and its cancers . K2P2.1  cancers . Further49 cells . Likewis49 cells .+ channels can be used to predict cancer cell response to anticancer drugs. The expression level of KCa1.1 channel is reduced in cisplatin-resistant ovarian cancer cells and its knockdown further promotes resistance to cisplatin [+ channels is positively correlated with chemoresistance, therefore K+ channel activators can enhance the therapeutic efficacy of conventional chemotherapy drugs. On the contrary, elevated expression of a few K+ channels has been shown to limit the efficacy of various chemotherapeutics. Therefore, inhibition of Kv10.1 promotes doxorubicin- and paclitaxel-induced cell death in breast cancer cell lines [+ channels may represent an effective therapeutic approach for improving cancer treatment.Moreover, the expression profile of Kisplatin . Similarisplatin . Reducedisplatin . Furtherisplatin . This evll lines . Likewisll lines . Togethe+ channels) are crucial for membrane excitability and cell communication. Depending on their mode of activation, two distinct classes of Na+ channels have been described, voltage-gated sodium channels (VGSC or NaV channels) which open in response to changes in membrane voltage, and ligand-gated sodium channels (LGSC or NaL channels) which are activated by the binding of specific ligands. The following sections focus on the significance of Na+ channels in cancer progression and their impacts on chemoresistance.Sodium channels , each of which contains six transmembrane segments (TMSs)  . The fou tissues . In addipoptosis .V expression is often upregulated, therefore inhibitors of NaV channels have been shown to decrease cancer cell invasion [V1.1 and NaV1.3 are highly expressed in ovarian cancer cells, while NaV1.2 and NaV1.4 are predominantly overexpressed in highly metastatic ovarian cancer cells compared to low-metastatic cells [V1.5 is increased in several cancers, including ovarian [V1.6 was observed in primary cervical cancer cells, and its blockade inhibited invasion of those cells [V1.7 induces growth and invasion of prostate [V channels is altered in various cancers. For instance, the expression level of the β1 subunit is negatively correlated with breast cancer cell migration, hence its overexpression promotes cell adhesion and reduces migration of MDA-MB-231 cells [In cancer, Nainvasion . For exaic cells . Further ovarian , colon [ ovarian , and bre ovarian , therefo ovarian . Similarse cells . Furtherprostate , gastricprostate , and endprostate  cancer c31 cells . On the 31 cells . A simil31 cells . This ex31 cells , breast 31 cells , and cer31 cells  cancers,31 cells .V channels has been also indicated to be associated with the chemosensitivity of cancer cells [V inhibitor lidocaine enhances the inhibitory effects of cisplatin on breast tumour metastasis and suppresses in vivo formation of lung colonies [V channels sensitizes breast cancer to taxol [V channels can differentially alter cell responses to anticancer drugs [Furthermore, the functional expression of Naer cells . For inscolonies ,275. Lidcolonies . On the to taxol . Likewiser drugs .L channels are activated by the binding of specific ligands. H+-NaL and Ca2+-NaL channels are the two well-known examples of NaL channels. H+-NaL channels are a group of voltage-insensitive Na+ channels called acid-sensing ion channels (ASICs). Currently, eight subunits of ASICs have been described: ASIC1a, ASIC1b1, ASIC1b2, ASIC2a, ASIC2b, ASIC3, ASIC4, and ASIC5. Each subunit consists of two transmembrane segments that assemble into homo or heterotrimeric complexes around a central pore [Naral pore . ASICs aral pore . Upregulral pore . Furtherral pore . Increasral pore . In brearal pore . TogetheFurthermore, ASIC1a is highly expressed in 5-FU- and doxorubicin-resistant hepatocellular cancer cell lines (Bel7402/FU and HepG2/DOXO) compared to their parental cells (Bel7402 and HepG2). Hereby, inhibition of ASIC1a by amiloride sensitizes Bel7402/FU and HepG2/DOXO cells to 5-FU and doxorubicin, respectively. Furthermore, exogenous overexpression of ASIC1a in Bel7402 and HepG2 cell lines promotes resistance to 5-FU and doxorubicin. These findings suggest that inhibitors of ASIC channels may serve as potential anticancer drugs by improving chemotherapy .2+-NaL channels are TRPM4 and TRPM5, which are monovalent-selective ion channels highly permeable to Na+. These channels open in response to increased intracellular Ca2+ levels. The altered expression profile of TRPM4 and TRPM5 has been associated with the etiology of several cancers. For instance, TRPM4 is increased in cervical and prostate cancers, and its downregulation reduces cancer cell proliferation and migration [2+-NaL channels in human cancers.Two common Caigration ,287,288.igration . TogetheDespite advancements in cancer therapy, cancer remains the second leading cause of death worldwide. Nevertheless, over the last few decades, targeted therapy has played a substantial role in improving the overall survival of cancer patients. Since the late 1990s, several small molecules and antibodies raised against specific tumourigenic proteins have been developed and approved by the U.S. Food and Drug Administration (FDA). However, the lack of druggable targets, compounded by severe toxicity profiles, has imposed a significant roadblock for anticancer drug discovery. Hereby, a growing number of studies are being conducted in order to discover novel and effective therapeutic targets for different malignancies. Recently, profound evidence has elucidated that proteins embedded in mammalian plasma membranes may unlock the fundamental basis for understanding carcinogenesis and disarming chemoresistance. Importantly, most ion channels and pumps are located in the plasma membrane and may serve as accessible and druggable targets for cancer treatment. In this regard, several studies have revealed a direct link between the functional dysregulation of membrane transporters and cancer development. Thus, many membrane transporters have been established as potential therapeutic candidates for cancer treatment. A growing body of evidence suggests that modulation of the expression and function of membrane transport proteins not only impacts cancer progression but also alters the cytotoxic effects of chemotherapeutics in different cancers. Given the diverse mechanisms of chemoresistance operating in human malignancies, discovering new therapeutic targets to enhance the efficacy of chemotherapy is crucial for improving patient outcomes. In this review, we emphasized the strong correlation between membrane transport proteins and carcinogenesis by focusing on three main aspects . First, Here we presented evidence on the impact of dysregulated membrane transporters on cancer growth, apoptosis, migration, and response to chemotherapy drugs. However, the extensive studies on the biological roles of membrane transporters in cancer are contrasted by a massive lack of information about their intrinsic properties and structural diversity. Hereby, recognizing the intrinsic regulation, gating kinetics, and structural diversity of membrane transporters is a key step toward uncovering their fundamental impacts on cancer progression and chemoresistance. Furthermore, the therapeutic approaches used to target different transporters have been discussed here; however, the lack of specific and potent drugs that can target distinct membrane transporters limits the therapeutic potential of these proteins. More importantly, developing novel drugs targeting membrane transporters requires further understanding of the biological function and structure of these proteins. Therefore, a better understanding of their structure and function may provide greater insights into their role in cancer progression and treatment as well as pave the way for the development of novel anticancer drugs and improvement of current chemotherapy efficacy.