Cancer is one of the leading causes of death around the world. The current treatment strategies, especially chemotherapy, have significant side effects. As a result, researchers are investigating alternative treatment strategies, including the use of natural compounds, such as iridoids. Iridoids are compounds predominantly found in plants and, in some cases, insects that have diverse therapeutic uses. These compounds have been reported to have a promising role in the management of cancer. The rise of cancer cases has coincided with the urgent need for the development of potent chemical entities and/or modification of existing commodities to improve their efficacy. Increasing evidence suggests that cancer remains one of the leading causes of death globally, with colon cancer cases alone likely to rise exponentially by 2030. The exponential rise in cancer prevalence is largely attributable to the growing change toward a sedentary lifestyle and modern diets, which include genetically modified foods. At present, the prominent treatments for cancer are chemotherapy, surgery, and radiation. Despite slowing cancer progression, these treatments are known to have devastating side effects that may deteriorate the health of the patient, thus, have a low risk–benefit ratio. In addition, many cancer drugs have low bioavailability, thereby limiting their therapeutic effects in cancer patients. Moreover, the drastic rise in the resistance of neoplastic cells to chemotherapeutic agents is rendering the use of some drugs ineffective, thereby signaling the need for more anticancer chemical entities. As a result, the use of natural derivatives as anticancer agents is gaining considerable attention. Iridoids have the potential to form conjugates with other anticancer, antidiabetic, antileishmanial, and antimalarial drugs, which synergistically have the potential to increase their effects. Published studies have identified the role of iridoids, which, if fully explored, may result in cheaper and less toxic alternative/adjuvant cancer drugs. The subject of this article is natural and synthetic iridoid derivatives and their potential therapeutic roles as anticancer agents. Cancer is one of the leading causes of death globally, with a recurrent exponential increase in new cases annually. Out of 183 countries, 112 countries reported that cancer is either the first or the second major cause of death in individuals below the age of 70 . DespiteTo find alternatives for cancer management, phytochemistry research has recently increased its focus on natural compounds, such as iridoid derivatives and their potential role in numerous diseases, including cancer ,13. ThesAnisomotpha buprestoides, Phaedon amoraciae, Phaedon cochleariae, Gastrophysa viridula, Plagiodera versicolora, and ants Iriabmyrmex detectus [In most plants, iridoids are primarily found in angiosperm dicotyledons and, in some cases, monocotyledons, where they are present in some leaves, fruits, roots, and sprouts ,18,19. Idetectus .Iridoids have been extensively researched for their anti-tumor properties and therapeutic benefits in chronic conditions ,24,25. R1) and catalpol (2), which have been found to have diverse pharmacological profiles [1) and catalpol (2), making them promising chemical entities [At present, there are prominent examples of iridoids that have attracted significant attention in cancer research, such as aucubin -pyran monoterpenoid compounds ,52 deriv1), have a glycosidic bond as the key feature, which is mostly present at aglycone C-1 or C-11 hydroxyl. They can be further subdivided into five groups depending on the number of carbon atoms on the skeletal systems, which range from 8 to 10 and 14, and alkaloid-conjugated iridoid glycosides [Glycosidic iridoids, such as aucubin , catalpol (2), and antirrhinoside (3) which are mostly present in the following plant families: Scrophulariaceae, Vebananaceae, Bignoniaceae, Oleaceae, and Plantaginaceae [Iridoids are found in dicotyledon angiosperms and, in some cases, monocotyledons . The isoginaceae ,56,57. TNatural iridoids are obtained from medicinal plants and animals using different extraction and isolation techniques, while synthetic iridoids are derived from chemical reactions using different reaction pathways ,58. OverThe most prominent solvent systems usually used in the isolation of iridoids are methanol, ethanol, butanol, and water ,63,64. TThe TLC technique, though ancient and tedious, has long been used in the isolation of iridoids. The technique is primarily used in purification with solvents and the identification of retention factor values and functional groups present ,66. AddiO-sinapoylgeniposide (4), isolated from Gardenia jasminoides used for the prevention or treatment of nervous system degenerative disorders, such as dementia. The reported iridoid was isolated using silica gel and column chromatography techniques. For the elucidation of the structure, techniques comprising 1D and 2D nuclear magnetic resonance (NMR) and high-resolution electrospray ionization–mass spectrometry (HR-ESI-MS) are used [A considerable body of literature has reported the use of the column chromatography technique in the isolation of iridoids ,69,70. Tare used . Howeverare used .5) and B (6), along with known compounds. The iridoids were isolated using techniques such as HPLC analysis and silica gel column chromatography. The chemical structure of the isolated iridoids was elucidated using IR, electrospray ionization–mass spectrometry (ESI-MS), HR-ESI-MS, and 1D and 2D NMR [O-β-D-glucopyranosyltrifloroside (7) and 4‴-O-β-D-glucopyranosylscabraside (8), together with three known secoiridoids, including trifloroside (9), scabraside (10), and gentiopicroside (11) from the rhizomes of Gentiana scabra. Column chromatography techniques were used in the isolation and purification of some, while HPLC was used for purification [12), 6-O-β-d-glucopyranosylgentiopicroside (13), swertiamarin (14), gentiopicroside (11), and sweroside (15) from Gentiana triflora and Gentiana rigescens [HPLC, on the other hand, is more frequently used in the isolation of iridoids, and it is also essential in their purification and quantification . Xu et ad 2D NMR . A phytofication . The isoigescens .16), 2′-O-caffeoylloganic acid (17),2-O-p-hydroxybenzoylloganic acid (18), 2-O-trans-p-coumaroylloganic acid (19), and 2-O-cis-p-coumaroylloganic acid (20) isolated from whole plants of Gentiana loureirii. The six known iridoids were 7-ketologanin (21), loganic acid (12), sweroside (15), boonein (22), and isoboonein (23), which were isolated with three other known compounds. To successfully isolate iridoids, combined chromatography techniques, including silica gel and octadecylsilyl (ODS) CC, were used, while spectroscopic techniques, such as 2D NMR, were used in structural elucidation [Recently, more advanced and intricate methods have been developed, and these include droplet counter-current chromatography (DCCC), high-speed counter-current chromatography (HSCCC), gas–liquid chromatography (GLC), and capillary zone electrophoresis (CZE) ,76. Wu ecidation .The aforementioned isolation techniques use various extraction methods with varying degrees of success in isolating iridoids. As a result, the percentage yields and recovery rates are equally impacted.Valeriana jatamansi yielded 570 g of a semi-solid residue from the extracts. An amount of 520 g of the extract was suspended in deionized water and extracted using different solvents. Seven iridoid derivatives were further isolated from the extract and accounted for a total mass of 568.2 mg [24), catalpol (2), and ajugol (25), weighing 21.0 mg, 18.0 mg, and 24.0 mg respectively [Below, some of the extraction techniques are succinctly summarized to provide an understanding of the separation strategies and their respective effectiveness in isolating iridoids from plants. The extraction conducted by Maurya et al. (2020) reported that roots weighing 4.0 kg obtained from 568.2 mg . Similarectively . The dra26), 6β-hydroxy geniposide (27), geniposidic acid (28), geniposide (29), and crocetin derivatives, including crocin-1, crocin-2, crocin-3, and crocin-4, with the following respective yields in some fractions of 151.1 mg, 52.2 mg, 24.5 mg, 587.2 mg, 246.2 mg, 34., 24.4 mg, and 24.7 mg, with a percentage purity ranging between 91.7% and 98.9% [Fructus Corni weighing 1.0 kg yielded 45 g of the pure compound. Subsequently, the yield of 100 mg taken for further fractionation produced 12.6 mg of loganin (30), 5.9 mg of sweroside (15), and 28.5 mg of morroniside (31)  [29) catalpol (2) catalposide (32), and verproside (33), was obtained through speed counter-current chromatography [However, techniques such as the counter-current chromatography method and speed counter-current chromatography appear to be more effective not only at recovering the mass, but also in obtaining pure iridoids. For instance, a study by Wang et al. (2015) reported a fraction weighing 1000 mg, which resulted in the isolation of four iridoids, namely gardenoside (nd 98.9% . Anotherctively) . Similar loganin , 5.9 mg tography ,81.Researchers are not capitalizing on the advanced techniques used in the isolation and purification of iridoids. Most recent studies still incorporate tedious and less effective techniques, such as CC which still has significant shortcomings, such as lower purity, poor recovery rate, and requiring a lot of time . ResearcVeronica species, namely verminoside (34), amphicoside (35), and veronicoside (36), were cytotoxic against the Hep 2 cell line, with IC50 values of 128 µM, 340  µM and 153.3 µM respectively while acetylcatalpol (37), aquaticoside B (38) and C (39), catalposide (32), veratroylcatalposide (40), and verproside (33) had cytostatic effects against Hep 2 cell lines at 100 µg/mL. Furthermore, verminoside (34) also displayed notable toxic effects when tested against RD (IC50 = 70 µM) and L20B (IC50 = 103 µM) cell lines. All of the iridoids investigated were cancer-selective, as they were not cytotoxic in the Vero cell line .In vitro and in vivo studies have played a pivotal role in broadening our understanding of the pharmacological profile of iridoids. In addition, structure-elucidation techniques have greatly assisted in depicting the functional groups responsible for the pharmacological profile of iridoids . ResearcGenipa americana conducted by Neri-Numa et al. (2020) evaluated the anticancer properties of extracts containing geniposidic acid (28), gardenoside (26), genipin-1-β-gentiobioside (41), geniposide (29), 6″-O-p-coumaroyl-1-β-gentiobioside geniposidic acid (42), 6″-O-p-coumaroylgenipin-gentiobioside (43), genipin (44), 6′-O-p-coumaroyl-geniposidic acid (45), and 6′-O-feruloyl-geniposidic acid (46). The extracts were tested against ion glioma and breast cancer cell lines , MCF-7 (breast), NCI-ADR/RES (breast expressing the multiple-drug-resistance phenotype), 786-0 , NCI-H460 , PC-3 (prostate), HT-29 (colon), and K562 (leukemia). Among the two extracts tested for anticancer properties, the unripe genipap fruit, which mostly contained genipin (44), was found to have anti-proliferative and anticancer effects against both glioma and breast cancer cell lines, with sample concentrations required to achieve 50% inhibition of cell proliferation GI50 (μg/mL) of 29.6 and 29.7. The genipin (44) standard was observed to inhibit all cell lines evaluated [A recent in vitro study on an iridoid-rich extract from the genipap fruit (ripe and unripe) of valuated .Valeriana dioscoridis were dioscoridin A (47), 1-epi-bosnarol (48), 8-epi-deoxyloganin aglycone (49), dioscorin B (50), dioscorin C (51), 10-acetylpatrinoside (52), and 10,2′-diacetylpatrinoside (53). These compounds were investigated for anticancer activity against three human cancer cell lines of gynecological origin  using an MTT assay. Among the isolated compounds, it was reported that 1-epi-bosnarol (18) had the highest anticancer activities, while the others showed moderate anticancer activity  when compared with cisplatin  [Furthermore, another in vitro study evaluated seven isolated compounds in which four iridoids were previously unreported. The isolated compounds from the roots of ., 2018) .54), was isolated by Krohn et al. (2007) together with an already known iridoid glucoside, gaertneroside (55). The isolated compounds were then tested for anticancer properties against MCF7 (breast cancer), NCl-H460 (lung cancer), and SF-268 (CNS cancer). When tested for anticancer properties in 60 human tumor cell lines, prismatomerin (54) was found to have high inhibition in leukemia cell lines (GI50 < 10 nM) [An unreported iridoid, prismatomerin (< 10 nM) .56) obtained from Citharexylum spinosum reported some promising anticancer properties. The synthetic analogues were synthesized by reactions including silylation, acetylation, and decinnamoylation. While 13 durantoside I analogues were synthesized, only two compounds were found to be more cytotoxic when compared with durantoside I (56) in all of the cell lines. A structural comparison of the compounds showed that the silyl-tert-butyldiphenylsilyl compound devoid of the cinnamate group was more cytotoxic than the compound with the cinnamate group at carbon 7 [A study by Apisornopas et al. (2018) on the synthesis of cytotoxic compounds from the natural iridoid glycoside (durantoside I) (carbon 7 .57), a natural iridoid isolated from Momordica charantia, was evaluated in vitro against leukemic (NB4 and K562), breast cancer (T47D) and liver cancer (C3A), normal human fibroblast (HF), and lung cancer (A549) cell lines. The results obtained demonstrated that plumericin (57) displayed considerable antiproliferative activity against two leukemia cell lines with effective doses (ED50) of 4.35 ± 0.21 and 5.58 ± 0.35 μg/mL [The anticancer efficacy of plumericin (35 μg/mL .58), 8-acet-ylharpagide (59), and harpagide (60), from Ajuga decumbens. Among the three isolated compounds, 8-acet-ylharpagide (59) had significant anticancer effects on EBV-EA induction and on mouse tumorigenesis. Additionally, 8-acet-ylharpagide (58) was evaluated for antitumor effects on mouse hepatic tumors, where 8-acet-ylharpagide (59) in the test groups was administered orally before and after treatment and for a period of 20 weeks [59) exhibited remarkable inhibitory effects in the two-stage carcinogenesis test.Another study that sought to identify anticancer compounds isolated three iridoids, i.e., reptoside (20 weeks . From th1), scandoside methyl ester (61), geniposide (29), loganin (30), sweroside (15), gardenoside (26), and gentiopicroside (11), were investigated for their anticancer properties. All of the iridoids demonstrated no anticancer activity. However, the aglycones of the iridoids, especially aucubin (1) and scandoside methyl ester (61), had significant anticancer activity against leukemia P388, with maximum total/control values of 162% and 160%, respectively, at 100 mg/kg. Scandoside methyl ester (61) was also evaluated against Ehrlich ascites carcinoma, meth A, sarcoma 180, and L1210, and was found to be more potent than 5 Fluorouracil [An in vitro and in vivo study on mice with leukemia cancer by Isiguro et al. (1986) evaluated iridoids (glycosides and aglycones). Iridoid glycosides, namely aucubin (64), mostly present in olives was reported by Hamdi et al. (2005) to have potent anticancer properties. In this study, oleuropein (64) was exposed to cancer cells including TF-1a, erythroleukemia; 786-O, renal cell adenocarcinoma; RPMI-7951 and RPMI-7951 melanoma cells; and LN-18 glioblastoma. Oleuropein (64) exhibited dose-dependent inhibition of advanced-grade tumor cell line growth and migration. Additionally, Oleuropein (64) irreversibly rounded cancer cells in a unique tube-disruption experiment, inhibiting their reproduction, motility, and invasiveness. In mice, 1% oleuropein (64) was observed to induce tumor regression in 10 mice out of 11 when administered orally between 9 and 12 days [A secoiridoid, oleuropein ( 12 days .Cerbera odollam in aqueous form. Ten semi-synthetic derivatives were tested for their cytotoxicity against SKBR3 (breast), HeLa , A375 (skin), HepG2 (liver), and HCT-116 (colon). Two derivatives, namely theveside-N-(p-methoxyphenyl)-carboxamide (65) (IC50 = 190 µM) (HCT-116), and theveside-N-(cyclohexyl)-carboxamide (66) (IC50 = 150 µM) (A375 cells), displayed moderate dose-dependent cytotoxicity in HCT-116 and A375 cell lines, respectively. However, theveside 2 (67)  and its derivatives, exhibited no disruption of cell viability in some cell lines , liver (HepG2), and breast (SKBR3)). The cytotoxicity of theveside 2 (67) was ascribed to cyclohexane as compared with the other derivatives that had aliphatic acyclic and aliphatic cyclic amides [Another phytochemical study reported the isolation of known iridoids from the extracts of leaves of c amides .68), in which the evaluation of in vitro and in vivo anticancer properties was conducted. The outcome of the study showed that Picroside II (68) exhibited anti-metastatic and anti-angiogenic properties against human breast MDA-MB-231 cancer cells. When administered at 20 and 40 μM, Picroside II (68) displayed considerable cell migration suppression at 66.67 ± 6.32% and 44.44 ± 5.67%. Additionally, picroside II (68) suppressed matrix metalloproteinase 9 and downregulated a cluster of differential 31 expression in cancer cells, and angiogenesis was significantly suppressed in the chick embryochorioallantoic membrane [Lou et al. (2019) published a study on iridoid glycoside (Picroside II) (membrane .69) demonstrated that the compound reduced the cell viability of breast cancer cells with no toxicity in a concentration-dependent manner, inducing apoptosis and cell cycle arrest. In vivo, Jatamanvaltrate p (69) was administered at a dose of 15 mg/kg in mice, where a significant reduction in cancer with no toxicity was noted, with 49.7% inhibition [A study on Jatamanvaltrate p (hibition .2) (>98%), geniposide (29) (99%), geniposidic acid (30) (> 98%), and harpagoside (70) (99%) and their hydrolyzed forms through the action of β-glucosidase investigated their potential anticancer properties in human prostate (DU145), breast cancer cells (MDA-MB-231), and multiple myeloma (U266) was conducted. All of the five iridoids that were not hydrolyzed showed no anticancer activity while the hydrolyzed iridoids significantly showed antitumor activity. Comparatively, hydrolyzed catalpol and hydrolyzed geniposide had high potency in all cancer cell lines [A study on iridoids (99.5%), catalpol (ll lines .Picrorrhiza kurroa extracts (PE) and their isolated iridoids, namely kutkin (71), picroside I (72), and kutkoside (73), against MCF-7 breast cancer cells. The research demonstrated that PE (IC50 = 61.86) and its isolated iridoids glycosides PS, KS, and KT had significant dose-dependent cytotoxic potential. Among the isolated iridoids, kutkin (71) displayed a considerable inhibitory activity of matrix metalloproteinases (MMPs) at 5 µM [An in vitro study was conducted by Rathee et al. (2013) on the anticancer activities of  at 5 µM .74) and 7-O-trans-cinnamoyl 6β–hydroxyloganin (75), which were derived from Nyctanthes arbortristis, and reported considerable anticancer activities. In the same study, chemical derivation was conducted, leading to the formation of two derivatives that exhibited an improvement in anticancer activities in HepG2 (human hepatocellular carcinoma) and MCF-7 (breast adenocarcinoma) [Pandeti et al. (2014) investigated iridoids, namely arbortristoside-A (rcinoma) .76) in SACC-83 cell lines, where the compound had an IC50 of 10 µM and hindered the development of the cell lines. The authors also observed that the cytotoxic effects of globularifolin (76) in healthy HGS cells were very minor, with an IC50 of 80 µM [11), which inhibits HeLa cells through multiple ways, including induced apoptosis, cycle arrest, and suppressed migration [Wang et al. (2019) reported the anticancer activities of globularifolin , and geniposide (29), were found to cause DNA to selectively inhibit topoisomerase I without interfering with topoisomerase II [2) has been cited to slow cancer progression through the down-regulation of the PI3K-Akt signaling pathway in HCT116, inhibition of TGF-β1-induced cell migration in colon cancer, and reduction of the expression of cyclin D1 in human solid tumor cell lines [77) was predicted by molecular docking studies to interact with the Stat3 protein at Cys712. Additionally, valtrate (77) may result in a brief reduction in intracellular glutathione (GSH) levels and an increase in reactive oxygen species (ROS) [76) revealed that the compound exhibited anticancer activities through the induction of apoptotic cell death in SACC-83 cells [1), catalpol (2), geniposide (29), geniposidic acid (28), and harpagoside (70) that were hydrolyzed through the action of β-glucosidase reported considerable cytotoxicity in the hydrolyzed iridoids. It was also observed that the suppression of the proliferation of K562 cells by iridoids involved the suppression of the activation of signal transducer and activator of transcription-3 (STAT3) [Picrorrhiza kurroa. such as Kutkin (71), Picroside I (72), and Kutkoside (73) demonstrated the inhibitory activity of matrix metalloproteinases (MMPs) [Iridoids have been implicated in acting on different proteins or pathways involved in cancer progression. Two iridoids, aucubin  . A study83 cells . Another (STAT3) . STAT 3 s (MMPs) . Few stuThe literature regarding the safety aspects of iridoids is currently sparse, as only a few studies have been published on that subject. In general, the profiles of the reported iridoids appear to be safe, and below are some of the few reported cases.Valeriana jatamansi orally administered at a dose of 3200 mg/kg body weight for an acute test. It was observed that, after 14 days, the extracts were exceedingly safe, there could be no single-dose harm, and no significant body changes were observed (p > 0.05). In a 90-day sub-chronic study, there were no observed adverse effects at a level of 1200 mg/kg/day. Additionally, the hematological and blood biochemical indicators and organ coefficients of the examined rats had no direct link with the toxicity of Valeriana jatamansi [A study by Xu et al. (2015) was conducted on an iridoid-rich fraction extracted from atamansi .29) administered at 320 mg/kg mg/kg, the effects of which were compared with those of 80 mg/kg genipin (44). In the study, acute toxicity was also determined by evaluating the liver weight and serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TB), and alkaline phosphatase (ALP) levels, and it was concluded that the conversion of geniposide (29) to genipin (47) was likely to be responsible for liver toxicity [A hepatotoxicity study was carried out by Yamano et al. (1990) in rats with the iridoid geniposide (toxicity .1) highlighted that the present data (in vivo and in vitro) were not sufficient to determine the safety aspects of aucubin (1). Most of the reviewed studies used mice/rats, and the reported safety concerns were movement reduction and food consumption. Despite the claims, most of the reported studies have highlighted that aucubin (1) did not seem to cause significant toxic effects [A review by Zeng et al. (2020) on aucubin ( effects .76) also improved the expression of Bax and Caspase 3 and 9, while decreasing the expression of Bcl-2. The SACC-83 cells were also inhibited by globularifolin (76) at the G0/G1 phase of the cell cycle. Additionally, a cell invasion assay demonstrated that globularifolin (76) prevented the migration of SACC-83 cells in a concentration-dependent manner, which was similarly associated with the downregulation of MMPs 2 and 9. The JAK/STAT pathway is crucial for the growth and development of cancer cells, and this study reported that globularifolin (76) might suppress this pathway [Treatment with globularifolin , scyphiphin D (78), ulmoidoside A (79), and ulmoidoside B (80). The six compounds were investigated at a concentration range of 2.4 to 40 µM and the results demonstrated that there was no significant toxicity when the compounds were exposed to RAW 264.7 cells. The cell viability was found to be greater than 94.15% [A study was conducted by Ma et al. (2022) on six iridoids isolated from the aerial parts of Viburnum luzonicum. The results of the cytotoxicity assay, which used the HeLa S3  cell line, demonstrated that two iridoid glucosides, namely luzonoside A (81) and luzonoside B (82) and their aglycons, exhibited moderate inhibitory activity, with IC50 values of 3–7 µM, whereas other isolated compounds showed no cytotoxicity, even at 100 µM [Nyctanthes arbortristi. The in vitro cytotoxic activity of Arbortristoside-C (83) was found to be 100 mg/mL against Hep-2 cells  and was compared with the control culture for each concentration [84) isolated from Cistanche phelypaea on human A431 squamous carcinoma cells and Hacat keratinocytes. The collected data showed that acetonide (84) was not harmful to human keratinocytes at the doses studied, but that it caused minimal cell death in tumor cells (between 12 and 20%) at the concentration of 100 M [Different findings have been reported regarding the cytotoxicity of iridoids. While a majority seem to be selectively cytotoxic, other iridoids have been observed to be potentially toxic. Fukuyama et al. (2004) isolated iridoids from t 100 µM . Anotherntration . Anotherof 100 M .50/EC50, it is still difficult to fully compare the anticancer properties of iridoids that have been reported in different studies. Given that the majority of iridoids are derived from plants, choosing an isolation process is crucial as it will surely alter the yield percentage and, if not selected carefully, could pose a serious threat to species that are vulnerable to extinction. The production of synthetic compounds also requires meticulous attention, as their safety profiles still lack in many aspects. Additionally, there are still gaps in the literature regarding the mechanism of action, structure–activity relationship, in vivo adverse effects, informatics, and pharmacophore models of most iridoids. Future studies should consider the safety aspects of iridoids, especially in clinical studies, and the mechanisms of action of iridoidsFrom the reported studies, most iridoids that act as anticancer agents are from plants, and this raises concerns regarding sustainability issues and ease of regrowth. It is, therefore, crucial to consider means of obtaining plant parts sustainably without posing any significant threat to the already endangered species. However, iridoids represent a promising group of new anticancer agents; thus, more bibliometric analysis, molecular docking studies, and clinical trials are required. The current reporting format of results in studies regarding the anticancer activities of iridoids (in vitro and in vivo) varies and, though some studies use ED


To explore clinical features and prognostic value of vascular endothelial growth factor (VEGF), interleukin (IL) 8, IL-10, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), and receptor-interacting protein-2 (RIP2) in diffuse large B-cell lymphoma (DLBCL).  A total of 68 DLBCL patients admitted to the Affiliated Hospital of Hebei Engineering University from January 2017 to June 2021 were included in this retrospective analysis. Serum VEGF was detected by enzyme-linked immunosorbent assay, serum IL-8 and IL-10 were detected by chemiluminescent enzyme immunoassay, and expression of PIK3CA and RIP2 in tumors was detected by immunohistochemistry. The correlation between clinical features of DLBCL and tumor-related index were analyzed. Cox regression was conducted to explore risk factors and hazard ratio. P < 0.05). Beside, these serum indexes were significantly higher in patients with the presence of extranodal involvement and germinal center B-cell (GCB), but significantly lower in patients with the presence of bone marrow involvement . Cox regression analysis for overall survival revealed that high expression of VEGF, high level of serum IL-8, serum IL-10, and RIP2, Ann Arbor Stage (III-IV), number of extranodal sites (>1), serum LDH level (≥245 U/L), IPI scores (3–5), ECOG scores (≥2), and bone marrow involvement were independent risk factors for the prognosis of DLBCL patients .  The serum level or expressions of VEGF, IL-8, IL-10, and RIP2 were significantly elevated with the increase of Ann Arbor Stage, International Prognostic Index (IPI) scores, Eastern Cooperative Oncology Group (ECOG) scores, serum lactate dehydrogenase (LDH) level, and the number of extranodal sites (all  The serum levels of VEGF, IL-8, and IL-10, as well as the expression of RIP2 and PIK3CA in tumor tissues, were highly correlated to clinical features of DLBCL, and high expression level of these indexes may have adverse effects for the prognosis of DLBCL patients. Non-Hodgkin lymphoma is one of the most highly heterogeneous malignant neoplasms in the lymphohematopoietic system, with a 12th place of incidence and a 10th place of mortality among malignant neoplasms . InternaκB pathway, which plays an important role in the pathogenesis of DLBCL 061). Informed consent was obtained from all the study subjects before enrollment.This retrospective study enrolled 68 patients with DLBCL admitted to our hospital from January 2017 to June 2021. Patients meeting the following criteria were included: (I) diagnosed according to the Chinese Guidelines for the Diagnosis and Treatment of DLBCL (2013 version)  and (II)The following characteristics of patients were included: age of DLBCL onset, gender, Ann Arbor Stage, B symptoms, the number of extranodal sites, serum level of lactate dehydrogenase (LDH), IPI scores, Eastern Cooperative Oncology Group (ECOG) scores, bone marrow involvement, extranodal involvement, and pathological types (Hans).Totally 5 mL of fasting venous blood of DLBCL patients was used for testing before treatment. VEGF was determined by the enzyme-linked immunosorbent assay , and IL-8 and IL-10 were determined by chemiluminescent enzyme immunoassay .Tissue specimen sections were placed in a 65°C oven for 1 h, dewaxed in xylene, dehydrated in gradient alcohol, and rinsed 3 times. According to the instruction of the immunohistochemistry kit , sections were incubated in 3% hydrogen peroxide solution for 10 min under dark conditions and washed by phosphate buffer saline (PBS), and antigen was repaired and incubated with a PIK3CA primary antibody or RIP2  primary antibody with a resolution of 1 : 200, thereafter incubated with a secondary antibody. Then, the sections were washed by PBS again, color developed using diaminobenzidine (DAB) solution, stained using hematoxylin, dehydrated, and mounted.Result evaluation: ten randomly selected high magnification fields in each section were observed under light microscopy by two senior physicians using a double-blind method, and the final results were averaged. PIK3CA results were determined by scoring the staining intensity into unstained (0 score), light yellow (1 score), dark yellow (2 scores), and brown (3 scores) and scoring the percentage of positive cells into ≤1% (0 score), 1%–20% (1 score), 21%–50% (2 scores), and >50% (3 scores). The positive PIK3CA was defined as positive, when the sum of these two scores ≥4, and as negative, when <4. RIP2 is located in the cytoplasm or around the nuclear membrane and is defined as positive when >40%.st 2022.Follow-up was performed by the phone or WeChat APP, and the last follow-up time was January 31, 2022. None of these 68 patients were lost to follow-up. The primary endpoint was overall survival and the second endpoint was disease-free survival. The starting point was set as receiving chemotherapy after diagnosis. The end point of overall survival (OS) was all cause death. The last follow-up time was January 31n (%) and tested by Chi-square test. Cox proportional hazard regression was conducted to explore risk factors and hazard ratios. All variables with P < 0.20 in univariate analysis were enrolled in multivariate analysis. P < 0.05 was considered as statistically significant.SPSS 23.0 software was used to perform statistical analysis. Measurement data were expressed as mean ± standard deviation (SD) and test by one-way ANOVA or LSD method. The categorical data were expressed as A total of 68 patients with DLBCL were included in this study , with a P > 0.05). There was also no difference of PIK3CA among subgroups, including bone marrow involvement, extranodal involvement, serum LDH level, and number of extranodal sites (P > 0.05). The serum level or expression of VEGF, IL-8 and IL-10 were significantly elevated with the increase of Ann Arbor Stage, International Prognostic Index (IPI) scores, Eastern Cooperative Oncology Group (ECOG) scores, serum lactate dehydrogenase (LDH) level, and the number of extranodal sites . Beside, these serum indexes were significantly higher in patients with the presence of extranodal involvement, but significantly lower in patients with the presence of bone marrow involvement . The number of patients with positive RIP2 and PIK3CA were significantly higher in patients with more extranodal site, serum LDH level ≥245, high IPI score, ECOG score ≥2, absence of bone marrow involvement, and extranodal involvement   cases with positive VEGF, 44 (64.71%) cases with positive IL-8, 43 (63.42%) cases with positive IL-10, 41 (60.29%) cases with positive PIK3CA, and 42 (61.75%) cases with positive RIP2. There was no difference of VEGF, IL-8, IL-10, PIK3CA, and RIP2 among different subgroups, including age, sex, histology type, and symptoms ( < 0.05) .The mean follow-up time was 39.87 ± 9.23 (24 to 54) months.P < 0.001).The OS of VEGF low expression and high expression groups were 33.12 and 14.28 months, respectively. The OS of PIK3CA low expression and high expression groups were 36.87 and 17.76 months, respectively. The OS of IL-8 low expression and high expression groups were 39.81 and 19.38 months, respectively. The OS of IL-10 low expression and high expression groups were 39.87 and 19.47 months, respectively. The OS of RIP2 low expression and high expression groups were 37.91 and 16.28 months, respectively , number of extranodal site >1 , serum LDH level ≥245 U/L , ECOG scores ≥2 , the bone marrow involvement , extranodal involvement , high IL-8 level , high IL-10 level , high VEGF level , positive PIK3CA , and positive RIP2  were risk factors for prognosis of DLBCL patients.As shown in P < 0.20 in univariate analysis were enrolled into multivariate analysis. Multivariate Cox regression revealed that Ann Arbor Stage III-IV , number of extranodal site >1 , serum LDH level ≥245 U/L , IPI scores 3–5 , ECOG scores ≥2 , bone marrow involvement , high expression level of IL-8 , high expression of IL-10 , high expression VEGF , and positive RIP2  were all independent risk factors for the prognosis of DLBCL patients , suggesting the possible connections of VEGF, IL-8, and IL-10 levels to the above clinical features. Further Cox proportional hazard regression analysis of prognosis showed that that Ann Arbor stage (stage III-IV), the number of extranodal site (>1), serum LDH level (≥245 U/L), IPI scores (3–5), ECOG scores (≥2), and the bone marrow involvement were risk factors for the prognosis of DLBCL patients. More importantly, high expression of VEGF, IL-8, IL-10, and positive PIK3CA and RIP2, were also risk factors for the prognosis of DLBCL patients, and the possible mechanisms are as follows: firstly, angiogenesis is extremely important for the growth, malignancy and metastasis of DLBCL. VEGF, which can act on vascular endothelial mitosis and induce the neovascularization formation, is the most potent cytokine contributing to tumor angiogenesis [κB pathway in human Burkitt lymphoma cells, which may be one of the mechanisms by which RIP2 affects the prognosis of DLBCL patients [In this study, the results showed significant differences of VEGF, IL-8, IL-10, and RIP2 between subgroups for Ann Arbor Stage, IPI scores, ECOG scores, the bone marrow involvement, extranodal involvement, serum LDH level, and the number of extranodal site (all ogenesis  and is aogenesis . Thirdlyogenesis . Finallypatients .There were also some limitations in this study. First, the sample size of this study was small and not representative enough, and the sample size should be expanded and the follow-up period should be extended in the future study. Second, the main tissues tested in this study for IHC were tumor tissues, and paraneoplastic tissues were not included to compare the differences of VEGF, IL-8, IL-10, PIK3CA, and RIP2. Third, this study was a retrospective analysis, and a prospective study should be used in the future to further confirm the results.In conclusions, advanced Ann Arbor Stage, higher number of extranodal site, high serum LDH level, high IPI score, high ECOG score, bone marrow involvement, high VEGF, high IL-8, high IL-10, and positive RIP2 were all risk factors of DLBCL overall survival time.


Among the prepared catalysts, carboxyl-functionalized ionic polymer (PIMBr-COOH) displayed the highest catalytic activity (94% yield) in the benchmark cycloaddition reaction of CO2 and epichlorohydrin, which was higher than hydroxyl-functionalized ionic polymer , and far exceeded ionic polymer without HBDs groups . Furthermore, PIMBr-COOH demonstrated good recyclability and wide substrate tolerance. Under ambient CO2 pressure, a number of epoxides were smoothly cycloadded into cyclic carbonates. Additionally, density functional theory (DFT) calculation verified the formation of strong hydrogen bonds between epoxide and the HBDs of ionic polymers. Furthermore, a possible mechanism was proposed based on the synergistic effect between carboxyl and Br− functionalities. Thus, a facile, one-pot synthetic strategy for the construction of bifunctional ionic polymers was developed for CO2 fixation.The development of bifunctional ionic polymers as heterogeneous catalysts for effective, cocatalyst- and metal-free cycloaddition of carbon dioxide into cyclic carbonates has attracted increasing attention. However, facile fabrication of such polymers having high numbers of ionic active sites, suitable types of hydrogen bond donors (HBDs), and controlled spatial positions of dual active sites remains a challenging task. Herein, imidazolium-based ionic polymers with hydroxyl/carboxyl groups and high ionic density were facilely prepared by a one-pot quaternization reaction. Catalytic evaluation demonstrated that the presence of HBDs (hydroxyl or carboxyl) could enhance the catalytic activities of ionic polymers significantly toward the CO Despirategies ,4. Recenfixation ,6. As a ectivity ,8,9.2 molecules, effective catalysts are essential to overcome the large thermodynamic energy barrier. In this context, various catalytic systems, such as metal salts [2 cycloaddition during the past few decades [2 cycloaddition in recent years [For chemical conversion of the inert COal salts ,11,12, ial salts ,14,15, aal salts ,17,18, h decades ,20. Gene decades ,22,23. O decades ,25,26. R decades ,28. Thernt years ,30,31,322-affinity, have shown enormous potential as heterogeneous catalysts for CO2 fixation [− as counterions have been fabricated for cocatalyst and metal-free cycloaddition of CO2 into cyclic carbonate, owing to the fact that Br− can promote this conversion via nucleophilicity, attacking the β-carbon atom with little steric hindrance in the epoxide ring [− generally exhibit inferior catalytic activity. To address this problem, a number of PILs bearing HBDs as electrophile nucleophiles and Br− as nucleophiles have been developed. Previous studies have shown that PILs functionalized with HBDs, such as hydroxyl [2 cycloaddition [Poly(ionic liquids) (PILs), which represent a new class of charged polymers featuring high ionic density and excellent COfixation ,34. Duriide ring ,35,36. Nhydroxyl ,38,39,40hydroxyl ,42,43, ahydroxyl ,45, exhiaddition . However− anions were successfully prepared. Consequently, the as-prepared ionic polymer with a carboxyl group (PIMBr-COOH) showed high catalytic efficiency and nice reusability toward CO2 cycloaddition. On the basis of the experimental results and DFT calculation, a possible mechanism involving the synergistic effect of carboxyl and Br− functionalities was proposed.Herein, HBDs-functionalized ionic polymers were successfully constructed based on the polymerization of commercially available 1,3,5-tri(1H-imidazol-1-yl)benzene and 1,3-dibromo compounds using a one-pot quaternization strategy. Through this facile method, bifunctional PILs bearing high numbers of imidazolium ionic sites, different types of HBDs, and definite spatial positions of HBDs near BrAs depicted in 13C NMR spectra, FT-IR, XPS, TEM, SEM, SEM-mapping, N2 adsorption–desorption, and TGA, were carried out. As seen in 13C NMR spectra of PIMBr, PIMBr-OH, and PIMBr-COOH showed similar signal patterns, suggesting their similar chemical structure. The characteristic peaks at about 128, 113, and 105 ppm were attributable to C2 and C5, C3–4, and C1 atoms of the polymer molecules [To investigate the physicochemical properties of the prepared ionic polymers, a number of characterization analyses, including solid-state olecules ,35. In tolecules . Compareolecules . It was olecules ,48.−1, which were attributed to the stretching vibrations of C = C [+ bonds [−1 was related to the C-O vibration of -COOH groups [−1 presented in the three samples were attributed to the peak of the absorbed H2O. Indeed, the adsorption of atmospheric H2O was a common phenomenon in PILs due to their hygroscopic properties [The FT-IR spectra of the three samples in of C = C ,50 and C[+ bonds , respectH groups , indicatoperties ,53. Thus2O in the polymer network, which was consistent with the result of the FT-IR analysis. As seen in 3/2 and 3d5/2 binding energy of Br− [XPS was conducted to further analyze the chemical structure of the prepared polymers. As depicted in y of Br− . Therefo2 g−1, respectively, which were similar to those of ionic polymers synthesized by the quaternization reaction [2 g−1 that was higher than that of PIMBr-OH. The relatively low surface areas of the three polymers could be ascribed to their high ionic density and flexible propylene linkers. High ionic density led to strong electrostatic interactions between the polymer chains [The porous properties of the prepared polymers were measured by nitrogen physisorption at 77 K. As summarized in reaction ,60. PIMBr chains ,60, whil2 fixation. PIMBr and PIMBr-OH displayed irregular agglomeration structures of large particles, and the particle size was much greater than that of PIMBr-COOH. A representative TEM image of PIMBr-COOH . As summarized in − [With the target ionic polymers in hand, their catalytic performance was evaluated in the COzed in − ,63,64, wzed in − . To conf2 pressure, and reaction time, were investigated to optimize the reaction conditions. As illustrated in 2 pressure was also evaluated. As summarized in 2 pressure reduced from 1 MPa to 0.1 MPa. Notably, 77% yield and high selectivity of target carbonate were still observed at ambient CO2 pressure. The good catalytic performance under low CO2 pressure should be assigned to the high ionic density and porous structure of the PIMBr-COOH catalyst. Under ambient CO2 pressure, the effect of reaction time was also checked. As displayed in 2 pressure (0.1 MPa), and 6 h.Having the optimal catalyst in hand, the influences of reaction parameters, such as temperatures, CO2 cycloaddition.Apart from catalytic activity, the recyclability of the solid catalysts is also a crucial indicator for evaluating their potential in industrial applications. Therefore, the recycling performance of PIMBr-COOH was evaluated under optimal reaction conditions. At the end of each catalytic run, the solid catalyst was centrifugally separated, washed with ethanol, dried under a vacuum, and then directly engaged in the next catalytic cycle. As shown in 2 cycloaddition reaction, epoxides bearing different functional groups were tested. As shown in 2 smoothly to afford excellent yields of corresponding products in 6 h , ethanol, 3-bromo-2-(bromomethyl)propionic acid (98%), 1,3-dibromo-2-propanol (95%), and epoxides were of analyzed grades and purchased from Energy Chemical . TIB (98%) was obtained from Alchemy Co. Ltd. CO2 was obtained from a local manufacturer with a purity of 99.9%.1,3-Dibromopropane (98%), N,N-dimethylformamide (30 mL) in a 100 mL pressure-resistant tube. After replacing the interior air with N2, the tube was transferred to an oil bath, stirred, and heated under 100 °C for 24 h. Subsequently, the tube was removed from the oil bath and cooled naturally to room temperature. Next, the resultant solid was washed thoroughly with ethanol, filtered, and dried under vacuum (80 °C) for 24 h. Finally, 1.03 g of white solid (98% yield) were obtained and named PIMBr-COOH (1.03 g). Following the same synthesis procedures of PIMBr-COOH, PIMBr and PIMBr-OH were also successfully fabricated in nearly quantitative yields by taking the place of 3-bromo-2-(bromomethyl)propionic acid with other 1,3-dibromo compounds, i.e., 1,3-dibromopropane and 1,3-dibromo-2-propanol, respectively.HBDs-functionalized PILs were fabricated via a one-pot quaternization strategy. In detail, TIB  and 3-bromo-2-(bromomethyl)propionic acid  were added to 2 cycloaddition, catalysts  and epoxide were successively added to a 25 mL high-pressure reactor equipped with a magnetic stirrer. After purging with CO2 several times, the reactor was charged with CO2 to 1 MPa and reacted at a pre-set temperature for the required time. After reaction completion, the system was cooled to room temperature. The resultant mixture was separated by centrifugation, and the obtained liquid was analyzed by GC and GC-MS using dimethylacetamide as the internal standard.In a typical reaction of catalytic CO13C NMR measurements were analyzed on a Bruker Avance 400 NMR spectrometer  with a 4 mm rotor spinning module. FT-IR measurements were recorded on a Nicolet Nexus FT-IR spectrometer  using the KBr pellet technique over a range of 4000–400 cm−1 at 4 cm−1 resolution and 32 scans. X-ray photoelectron spectroscopy (XPS) analyses of the prepared PILs were performed on a Thermo Fisher Scientific K-Alpha spectrometer  equipped with Al K radiation (1486.68 eV). Porous properties of the prepared PILs were measured at 77 K on a Micromeritics ASAP2460 after degassing at 100 °C for 8 h. A scanning electron microscope  with energy-dispersive X-ray spectroscopy (EDS) and a transmission electron microscope , operating at a high voltage of 200 kV, were used to investigate the morphology of the materials. Thermogravimetric analysis  was carried out using a NETZSCH STA 449F3 over a temperature range from 25 to 600 °C at a heating rate of 10 °C/min under a dynamic N2 atmosphere. GC and GC-MS analyses were recorded on SCION 456-GC and Agilent 6890/5973 GC-MS, respectively.Solid-state DFT calculations were performed to investigate the hydrogen-bond interactions between epichlorohydrin and designed catalysts using the Gaussian 09 package. The geometry of all molecules was optimized by the B3LYP-GD3 level using the 6-31G*  basis se2 into epoxides. Carboxyl-functionalized ionic polymers displayed higher catalytic activity than those of HBDs-free and hydroxyl-functionalized ionic polymers. DFT calculation demonstrated that the formation of hydrogen bonds between epoxide with HBDs of ionic polymers facilitated the ring-opening step, and the higher catalytic activity of PIMBr-COOH was mainly attributed to its stronger hydrogen bond interaction with the epoxide substrate. Additionally, PIMBr-COOH exhibited good recyclability and was able to be reused at least five times. Under ambient CO2 presence, a series of substituted epoxides were smoothly transformed into cyclic carbonates in excellent yields under solvent-, cocatalyst-, and metal-free reaction conditions. This research thus represents a reliable strategy for direct fabrication of carboxyl-functionalized ionic polymers towards efficient CO2 conversion.In summary, novel ionic polymers with different types of HBDs, high numbers of imidazolium ionic sites, and controlled spatial positions of dual active sites were facilely constructed by a one-pot quaternization strategy. Impressively, the presence of hydroxyl and carboxyl groups boosted the catalytic activities of ionic polymers significantly toward the cycloaddition reaction of CO


C2, at-risk haplotype MCPggaac, and excellent response to therapeutic complement inhibition, points to the pivotal role of complement in the pathophysiology of disease. Moreover, the importance of a multidisciplinary team approach in  thrombocytopenia is emphasized.Postsurgical thrombotic microangiopathy (TMA) is a complication associated with significant morbidity and mortality. Still, the pathophysiological underlying mechanism of postsurgical TMA, a diagnosis often overlooked in postoperative patients with acute kidney injury and thrombocytopenia, is largely unknown. Here, we report the case of a 56-year-old male that developed anuric acute kidney injury, Coombs-negative hemolysis, and thrombocytopenia after surgical aortic arch replacement. Massive ex vivo complement activation on the endothelium, a rare complement gene variant in  Aortic surgery, especially in a re-operative setting, carries a significant early morbidity and mortality rate, ranging between 5 and 10% , a severPostsurgical TMA, although considered a rare entity, has been observed in up to 5% of patients following aortic surgery . The pat9/L), and Coombs negative microangiopathic hemolytic anemia , suggesting TMA. Coagulation tests were within the (pseudo)normal range , prothrombin time 15.5 s ), while overt bleeding was absent. Plasmapheresis was initiated immediately, awaiting the results of enzymatic activity of von Willebrand factor cleaving protease , which proved to be within the normal range . Routine complement measures showed low C4  and C3 . Moreover, serum induced massive ex vivo complement activation as determined by formation of the membrane attack complex  on microvascular endothelial cells ), indicating C-TMA [A 56-year-old male patient presented to our center with a residual dissection and progressive dilatation of the proximal descending aorta (68 mm) and other aortic segments A followiCFH, CFI, CD46, CFB, C3, CFHR1-5, C2, CFP, and THBD) demonstrated a rare variant in C2 c.841_849+19del, with a minor allele frequency of 0% (Exome Variant Server) to 0.4% (gnomAD), at-risk haplotype MCPggaac, and deletion of CFHR1 and CFHR3; all the genetic abnormalities came in heterozygosity. Factor H autoantibodies, however, were not found.DNA sequencing and multiplex ligation-dependent probe amplification of complement genes  C-TMA can be challenging and is frequently delayed because features of TMA overlap with several more obvious sequelae of surgery . For exaC2 to form the C3 convertase . Subsequently, the activation of C3 leads to the assembly of the C5 convertase, activating the terminal complement pathway via C5 and the formation of C5b9. Moreover, prosthesis-related hemolysis with the release of heme can activate complement on the endothelium [C2; recently, a gain-of-function C2 protein, a paralog of the alternative pathway protein complement factor B [MCPggaac decreases transcriptional activity of the CD46 promoter region, affecting complement regulation downstream of C2 [Most reports on postsurgical TMA do not provide data on complement measurements. To the best of our knowledge, two patients with postsurgical TMA presented with variants of uncertain significance in complement genes , suggestothelium ,18. At pothelium . DNA seqfactor B , that leam of C2 . We therC2 variants are warranted.Postsurgical TMA likely represents a mixture of distinct causes, only some of which may be associated with deregulated complement. Plasma exchange should be started immediately in patients with postsurgical TMA. Plasma exchange has been associated with clinical remission in 30% to 80% ,14, poinC2 variant combined with MCPggaac, and (iii) excellent response upon eculizumab. Furthermore, we emphasize a multidisciplinary approach of  TMA after cardiovascular surgery to optimize treatment for this life-threatening complication  massive ex vivo C5b9 formation on the endothelium, (ii) a novel lication .


VPS13A-D genes result in rare neurological diseases, including chorea-acanthocytosis. The pathogenesis of these diseases is poorly understood, and no effective treatment is available. As VPS13 genes are evolutionarily conserved, the effects of the pathogenic mutations could be studied in model organisms, including yeast, where one VPS13 gene is present. In this review, we summarize advancements obtained using yeast. In recent studies, vps13Δ and vps13-I2749 yeast mutants, which are models of chorea-acanthocytosis, were used to screen for multicopy and chemical suppressors. Two of the suppressors, a fragment of the MYO3 and RCN2 genes, act by downregulating calcineurin activity. In addition, vps13Δ suppression was achieved by using calcineurin inhibitors. The other group of multicopy suppressors were genes: FET4, encoding iron transporter, and CTR1, CTR3 and CCC2, encoding copper transporters. Mechanisms of their suppression rely on causing an increase in the intracellular iron content. Moreover, among the identified chemical suppressors were copper ionophores, which require a functional iron uptake system for activity, and flavonoids, which bind iron. These findings point at areas for further investigation in a higher eukaryotic model of VPS13-related diseases and to new therapeutic targets: calcium signalling and copper and iron homeostasis. Furthermore, the identified drugs are interesting candidates for drug repurposing for these diseases.Mutations in human  According to European Union law, a disease is considered rare when it affects no more than 1 person per 2000. However, with around 7000 identified rare diseases, together they affect 3.5–10% of the population, which corresponds to approximately 30 million people in Europe and 300 million around the world ,7,8,9. DAnother obstacle that the patients face is a great limitation of therapies. For a significant majority of rare diseases, current treatment is only symptomatic and focuses on improving the quality of patients’ life; however, the disease itself is not cured. Therefore, there is a great need to find new drug targets and drugs for rare diseases. Drug targets can be easily identified in yeast models by genetic methods, such as second-site suppressor screens and multicopy suppressor screens, which indicate where to hit to overcome observed defects. The costs of launching a new drug to the market may reach up to several billions of dollars, and the rare disease market is very limited . In ordeVPS13 (vacuolar protein sorting 13) genes using yeast as a model organism. Emphasis was given to recently published work on VPS13 proteins in the Special Issues of IJMS entitled “Yeast Models and Molecular Mechanisms of Neurodegenerative Diseases” and “Yeast Models and Molecular Mechanisms of Neurodegenerative Diseases 2021”.Here, we summarize attempts to improve our understanding of the pathogenesis and to identify potential therapeutic targets and repurposable drugs for rare diseases associated with mutations in S. cerevisiae yeast, despite a simple, unicellular structure, is widely used for studying human diseases. Yeast models of neurodegenerative diseases [ATP7A and ATP7B genes resulting in Menkes and Wilson’s diseases, respectively [diseases ,16,17, mdiseases ,19,20, mdiseases , ageing diseases ,23, priodiseases ,25 or evdiseases  were estdiseases  as well diseases , and somectively ,30,31—orectively ,18,20. Fectively .GDAP1 gene [Modelling human diseases in yeast is possible even in cases when there are no orthologs in both species. Heterologous expression of a human gene can still influence the functioning of yeast cells. Examples are yeast models of neurodegenerative diseases with protein aggregation. Expression of genes encoding aggregating protein results in toxicity. In this manner, yeast models of Alzheimer’s disease (AD), Parkinson’s disease (PD) and Huntington disease (HD) have been developed, in which respective aggregating proteins, amyloid β, α-synuclein and huntingtin, are produced ,33. In cAP1 gene . While yOnce a yeast disease model is established, one can apply various research approaches that aim to discover new aspects of a disease pathogenesis and contribute to its treatment. One of the experimental approaches used in yeast models is screening for suppressors. A suppressor is a genetic or chemical factor that overcomes defects caused by a mutation. Among genetic suppressors, two classes can be distinguished: second-site and multicopy suppressors . MulticoSuppression could be achieved by treating a yeast model with biologically active chemical compounds. This kind of suppression is referred to here as chemical suppression, and it is analogous to drug intervention in humans. Drugs that suppress the effects of the modelled mutation could be identified by screening chemical libraries . HoweverUsing yeast genetic and chemical screens has been fruitful in elucidating the pathogenesis of many diseases, and the findings in yeast models have been confirmed in higher eukaryotic disease models ,37,38,39VPS13 are evolutionarily conserved genes encoding VPS13 proteins; thus, it is possible to study them in various model organisms [S. cerevisiae yeast, where one VSP13 gene, encoding the Vps13 protein, was present. With a length of 3144 amino acid residues and a molecular mass of 358 kDa, Vps13 is one of the largest proteins found in yeast. It shows a complex domain structure, which is shared with human homologous proteins  homotypic fusion and TGN to multivesicular body transport [vps13Δ mutant shows elevated transfer of mitochondrial DNA to the nucleus and enhanced mitophagy [vps13Δ mutant also exhibits defects in the actin cytoskeleton organisation and endocytosis [Originally, tidase Y ,45. In froteases ,47, vacuroteases , sortingroteases , vesicleroteases . Sortingransport . Next toitophagy . The vpsocytosis , a proceocytosis . Moreoveocytosis  and is pocytosis . This suocytosis ,54,55.S. cerevisiae yeast was shown to bind phospholipids and transfer them between phospholipid bilayers in vitro [Chaetomium thermophilum fungus revealed that it forms a long groove with hydrophobic and hydrophilic amino acid residues facing the interior and exterior sites, respectively. This groove architecture is suitable for solubilizing lipids, suggesting that Vps13 serves as a bridge between the membranes of different organelles and mediates bulk lipid flow [Recent studies strongly suggest that VPS13 proteins act as lipid transfer proteins at membrane contact sites (MCS)—zones of close proximity between the membrane of one organelle and the membrane of another organelle or the plasma membrane. MCS enable the direct exchange of small metabolites , signal transduction and enzymatic regulation in trans . The N-tin vitro . Moreovepid flow ,59. It ipid flow . The recpid flow .2+) [Vps13 in yeast cells was found at the MCS between the vacuole and mitochondria; vacuole and nucleus; and endosome and mitochondria  51,62,6,662,63. 2+) . The inv2+) .Research regarding Vps13 protein using the yeast model contributed to a better understanding of Vps13 functioning and gave background to recent groundbreaking studies regarding VPS13 proteins’ role in lipid transfer. It is one of the many examples of how yeast as a model of a eukaryotic cell improved the state of knowledge in the field of cell biology.VPS13 gene family consists of four members: VPS13A, VPS13B, VPS13C and VPS13D [VPS13 genes ranges from 208 to 864 kb, and exons number from 66 to 86 [VPS13 genes are associated with several rare neurological diseases. VPS13A-D proteins localize to various MCS, where it is possible that they all transfer lipids between membranes.The human d VPS13D . The len66 to 86 . Express66 to 86 ,70,71. MVPS13A result in chorea-acanthocytosis (ChAc), an ultrarare neurodegenerative disease [VPS13A-delin mutation was reported [VPS13A knockout cell lines, such as abnormal mitochondrial morphology, increased mitochondrial fission and reduced elimination of damaged mitochondria by mitophagy [VPS13A, probably caused by impaired processing of lysosomal hydrolases [VPS13A gene manifest clinically similarly to mutations in the XK gene causative of McLeod syndrome [2+-dependent manner [Mutations in  disease ,71,72 af disease . In most disease ,75,76,77 disease . These s disease . Recentlreported . In addireported ,80. VPS1reported ,81,82. Ditophagy . This codrolases ,85. It idrolases , which adrolases ,88. Moredrolases . The altdrolases . Signifidrolases . Recentldrolases . Surprisdrolases ,93, but drolases . This isdrolases . Findingsyndrome . VPS13A syndrome , and it syndrome . Moreovet manner .VPS13B lead to Cohen syndrome—a disorder characterised by psychomotor retardation, intellectual disability, microcephaly and characteristic facial features [VPS13-related disease that is manifested already at birth. Similarly to ChAc, mutations in VPS13B predominantly result in the lack of functional protein [Mutations in features ,96. Cohe protein . VPS13B  protein . Localis protein , a GTPas protein ,101,102. protein . This de protein . Finally protein .VPS13C are associated with early-onset PD [VPS13C function results in abnormal mitochondrial morphology, lower mitochondrial membrane potential and increased respiration rates [Mutations in onset PD ,106,107 onset PD ,110,111.onset PD . Anotheronset PD . Loss ofon rates . Finallyon rates ,114,115.on rates .VPS13D result in complex neurological disease, named VPS13D movement disorder. Patients exhibit movement defects , in some cases combined with intellectual disability [VPS13D function causes reduced mitochondrial fission, resulting in defective mitophagy and impaired mitochondrial morphology [Mutations in sability ,117,118.rphology . Moreoverphology . The latrphology . Anotherrphology .VPS13A gene, which is most similar to yeast VPS13 [vps13Δ; therefore, the vps13-I2749R mutation mimicking vps13A-I2771R point mutation found in the ChAc patient was functionally analysed. This mutation causes amino acid residue substitution in the APT1 domain and abolishes Ca2+-dependent binding of this domain to selected phosphorylated phosphatidylinositols in vitro and changes Vps13 localisation in vivo [VPS13A-D mutations were tested in the same way in yeast: vps13A-L67P, vps13A-L1095P, vps13A-Y2721C; vps13B-N2993S, vps13C-W395C, vps13C-F444P, vps13C-L2789T, vps13D-L2900S, vps13D-N3521S, vps13D-D4107I, vps13D-A4210V and vps13D-R4228Q, [The st VPS13 , does no in vivo . Several-R4228Q, . The VPS-R4228Q, .The knowledge about VPS13 proteins is still not sufficient to develop a specific therapy for patients. To date, gene therapy is also not available. Only symptomatic treatment is used, and new methods to alleviate the symptoms are needed. The most efforts for treatment development are reported for ChAc. The deep brain stimulation is used to relieve movement problems , botulinVPS13-dependent diseases were constructed, which show various phenotypes [vps13, hypersensitivity to commonly used detergent sodium dodecyl sulfate (SDS), enable suppressor screens [To find therapeutic alternatives based on the understanding of cell biology, using model organisms is necessary. Several yeast models of enotypes ,13,48,51 screens . This gavps13-I2749R mutation, and several plasmids responsible for improved growth were isolated and analysed. One of them contained a fragment of the MYO3 gene (MYO3-N) encoding N-terminal part (amino acid residues (aa) 1–775) of Myo3 protein (Myo3-N) [MYO3-N overexpression was also improving growth of the vps13Δ mutant in the presence of SDS. Importantly, the suppression by MYO3-N was not limited to SDS-hypersensitivity. MYO3-N corrected two other vps13Δ defects: depolarisation of the actin cytoskeleton and hypersensitivity to canavanine, the phenotypes implying defective endocytosis. Indeed, endocytic reporters  that localise to sites of endocytosis, were shown to be present longer on the plasma membrane in the vps13Δ cells than in the wild-type, indicating delayed endocytosis. MYO3-N overexpression shortened patch lifetimes for Las17, Myo3 and Myo5. To conclude MYO3-N overexpression could partially improve endocytosis in vps13Δ alleviating canavanine hypersensitivity [Specifically, SDS hypersensitivity growth phenotype was used in a screen for multicopy suppressors of (Myo3-N) . Myo3 is(Myo3-N) ,128. MYOsitivity .MYO3-N action was elucidated. The suppressing MYO3-N fragment encodes myosin motor domain and a linker with two calmodulin binding motifs (IQ1 and IQ2). It is possible that suppression could be achieved by binding with calmodulin (Cmd1)—a conserved, calcium-binding protein mediating calcium signalling in cell [MYO3-N and resulted myo3-iq1/2 was not able to suppress vps13Δ and the interaction between Myo3-iq1/2 and Cmd1 was abolished. These results indicated that binding of calmodulin to Myo3-N is necessary for MYO3-N-based suppression of vps13Δ [vps13Δ cells than in the wild-type, and MYO3-N overexpression reduces it , which limits the calcium availability required for calmodulin and calcineurin activity, and FK-506  analysis. The other structural criteria established during SAR analysis implicated that the heterocyclic pyranic ring must contain a double bond between C2 and C3 atoms and a carbonyl group, yet the ring itself does not necessarily have to be closed. A similar case is tolcapone, where the criteria regarding hydroxyl and carbonyl groups are met and benzyl rings are not connected by heterocyclic pyranic ring but by a carbonyl group.There are not many reports on iron or copper dyshomeostasis in s so far . Findingf vps13Δ ,170 provpressors . Luteolipressors ,173,174.pressors ,175. Tolpressors , has sompressors , while ferformed . An addivps13Δ suppression overlaps with those previously described for flavonoids responsible for the antioxidant and metal chelation properties [vps13Δ was shown to be hypersensitive to cadmium [vps13Δ SDS hypersensitivity were also active when tested for vps13Δ cadmium hypersensitivity, a possible mechanism of their action could rely on their antioxidant properties. Contrary to this prediction, some of the tested flavonoids, such as kaempferol, did not improve vps13Δ growth, despite having higher antioxidant potential than the suppressing flavonoids [vps13Δ. Thus, it was hypothesised that metal chelation properties could possibly be important for vps13Δ suppression. This was supported by the fact, that one of the identified genes in the multicopy suppressor screen was FET4 [vps13Δ growth. These results highlight the importance of iron in the protection against SDS stress. The potential interaction between luteolin and FET4 overexpression was tested and luteolin improved vps13Δ growth regardless of FET4 overexpression, and no additive effects were observed [FET4  could be a target for copper-based suppression in s proved ,205,206.3Δ cells . Next togolipids ; howevergolipids . Therefovps13Δ mutant was particularly useful for identifying several multicopy and chemical suppressors. Importantly, some of the suppressors, both multicopy and chemical, were found to act on the same pathways relevant for vps13Δ suppression. Analysis of these pathways revealed new defects present in Vps13-deficient cells, showing that they could be potential therapeutic targets.Several studies show that yeast could be successfully used in the modelling of human neurological diseases to better understand their pathology, find therapeutic targets and repurpose drugs. The SDS hypersensitivity phenotype of the yeast vps13Δ cells. It was shown that the calcineurin activity increased in vps13Δ cells and the downregulating activity of Cmp2 catalytic subunit suppressed vps13Δ SDS hypersensitivity. Activity reduction could be achieved by overexpression of MYO3-N and RCN2, deletion of CMP2, or treatment with EGTA and FK-506. MYO3-N overexpression also corrected endocytosis and the defect in the actin cytoskeleton’s organisation in vps13Δ cells. These findings are especially interesting, because the actin cytoskeleton organisation is regulated by calcineurin [The new findings show the importance of calcineurin for the functioning of cineurin . Moreovecineurin . Epileptvps13Δ suppression. Overexpression of the FET4 gene or treatment with iron salts improved vps13Δ growth. Increased iron import could also be achieved in a copper-dependent manner. Moreover, flavonoids may act using the same process as iron. However, the exact mechanism in which iron improves yeast growth in the presence of SDS is still not elucidated. One possibility is that both iron and flavonoids, and perhaps copper itself, improve sphingolipid biosynthesis in vps13Δ. Sphingolipids are important for SDS protection in yeast and for nervous system functioning in humans. Therefore, knowing that VPS13 proteins influence lipid transfer in cells, disturbed sphingolipid homeostasis may be an important factor in the pathogenesis of the VPS13-related diseases. Future studies are required to determine whether sphingolipid homeostasis is disturbed in human cells from patients and could be a target for therapeutic intervention.The other findings show the relevance of iron, copper homeostasis, copper ionophores and flavonoids for siVPS13A [VPS13-related diseases.Newly identified potential repurposable drugs which are effective in alleviating defects of yeast mutant cells, such as calcineurin inhibitors, flavonoids and copper ionophores, require intensive studies using available human cell and mouse models. HeLa siVPS13A , fibroblsiVPS13A  togethersiVPS13A  will hel


Inflammation is a pivotal response to a variety of stimuli, and inflammatory molecules such as cytokines have central roles in the pathogenesis of various diseases, including bone and joint diseases. Proinflammatory cytokines are mainly produced by immune cells and mediate inflammatory and innate immune responses. Additionally, proinflammatory cytokines accelerate bone resorption and cartilage destruction, resulting in the destruction of bone and joint tissues. Thus, proinflammatory cytokines are involved in regulating the pathogenesis of bone and joint diseases. Interleukin (IL)-1 is a representative inflammatory cytokine that strongly promotes bone and cartilage destruction, and elucidating the regulation of IL-1 will advance our understanding of the onset and progression of bone and joint diseases. IL-1 has two isoforms, IL-1α and IL-1β. Both isoforms signal through the same IL-1 receptor type 1, but the activation mechanisms are completely different. In particular, IL-1β is tightly regulated by protein complexes termed inflammasomes. Recent research using innovative technologies has led to a series of discoveries about inflammasomes. This review highlights the current understanding of the activation and function of the NLRP3  inflammasome in bone and joint diseases. Proinflammatory cytokines are important molecules that mediate inflammatory and innate immune responses and are involved in the development of inflammatory and chronic diseases ,2,3, incIL-1α and IL-1β are known isoforms of IL-1 ,12. BothAlthough IL-1α and IL-1β have the same biological activity, their activation mechanisms are completely different ,12. IL-1IL-1α and IL-1β secretion is mediated by the same unique system using plasma membrane permeabilization ,17. BecaRecently, inflammasomes have been reported to be associated with the onset and progression of bone and joint diseases  as well Inflammasomes are molecular platforms for caspase-1 activation and are typically composed of three parts: a sensor protein, adapter protein, and pro-caspase-1  Figure . InflammNLRP3, NLR family CARD domain-containing protein 4 (NLRC4/IPAF), and absent in melanoma 2 (AIM2) are representative PRRs that form inflammasomes. The regulatory and activation mechanisms of NLRC4 and AIM2 are better understood compared with those of NLRP3.NLRC4 recognizes cytosolic flagellin (fliC), a structural protein in bacterial flagella, through the NLR family, apoptosis inhibitory protein (NAIP) 5/6  Figure . NAIP5/6AIM2 consists of a C-terminal HIN-200 domain and N-terminal PYD ,29 Figu. The HINNLRP3 gene have been reported to cause an autoinflammatory disease known as cryopyrin-associated periodic syndrome (CAPS), which shows systematic and joint inflammation with hyperactivation of the NLRP3 inflammasome [Unlike NLRC4 and AIM2, NLRP3 senses various stimuli, such as bacterial infection, extracellular ATP, crystals, lysosomal rupture, mitochondrial damage, and ER stress ,33 Figu. Howeverammasome ,35. TherActivation of the NLRP3 inflammasome requires a two-step process termed Signal 1 and Signal 2 ,33 Figu. Signal + efflux, Ca2+ signaling, mitochondrial damage, and reactive oxygen species (ROS) have been proposed as intracellular signals for NLRP3 inflammasome activation [+ efflux, Ca2+ signals, and ROS. These intracellular signals are mediated by at least the P2X7 receptor, pore-forming toxins, lysosomal rupture, Ca2+ channels, mitochondrial damage, ER stress, and G protein-coupled receptors (GPCRs) [+ efflux is the most important for NLRP3 inflammasome activation [+ efflux [NLRP3 contains PYD, NACHT, and leucine-rich repeat (LRR) domains . Becausetivation ,38,39,40 (GPCRs) ,33,41,42tivation , althoug+ efflux ,45.+ efflux [+ efflux and binds to NLRP3 remain unclear. Therefore, understanding the molecular mechanisms that link NEK7 to K+ efflux would facilitate elucidating the precise mechanism of NLRP3 inflammasome activation.NIMA-related kinase 7 (NEK7) is a critical regulator for NLRP3 activation downstream of K+ efflux ,47,48  has been reported to be an essential regulator for cell death induced by caspase-1 ,51 Figu. ActivatRegarding cell death induced by the NLRP3 inflammasome, DEAD-Box Helicase 3 X-Linked (DDX3X)  has beenBecause negative regulation of the NLRP3 inflammasome is a potential therapeutic target for NLRP3-associated diseases, negative regulators of the NLRP3 inflammasome have been investigated . In partAutophagy is an intracellular degradation system that delivers cytoplasmic material and organelles to the lysosome . Loss ofThe ubiquitin–proteasome system is also associated with the negative regulation of the NLRP3 inflammasome . UbiquitBecause DRD1 and TGR5 are GPCRs, G protein signals are also involved in the negative regulation of the NLRP3 inflammasome. GPSM3  suppresses NLRP3 inflammasome activation by interacting with the C-terminal LRR domain of NLRP3 . RalB , the non-canonical NLRP3 inflammasome pathway was discovered ,74. The + efflux, which triggers the activation of the non-canonical NLRP3 inflammasome for IL-1β and IL-18 maturation. NEK7 is required for the non-canonical NLRP3 inflammasome downstream of K+ efflux. Recent studies have shown that several guanylate-binding proteins (GBPs) bind to cytosolic LPS and are implicated in caspase-4 activation [In addition to caspase-11, human caspase-4 (caspase-11 in mice) and caspase-5 activate the non-canonical NLRP3 inflammasome pathway  Figure . These ctivation ,79,80,81In addition to cytosolic LPS, oxidized phospholipid 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (oxPAPC), which modulates inflammatory responses, has also been reported to bind to caspase-11 and activate the non-canonical pathway . CytoplaRA is a chronic autoinflammatory arthritis that leads to joint malformations and dysfunction . ProinflNLRP3 gene correspond to RA susceptibility [The NLRP3 inflammasome is involved in the pathogenesis and development of RA , and poltibility ,92,93,94tibility ,96, suggA20/TNFAIP3 has been found in monocytes from RA patients [A20 in mice triggers spontaneous arthritis [Various RA model mice have been analyzed in the field of inflammasomes. The NLRP3 inflammasome is activated in synovial tissues of collagen-induced arthritis (CIA) mice . Expresspatients , and myerthritis . NLRP3 drthritis . The CASrthritis . Anti-inrthritis . Howeverrthritis ,105. TheOA is a progressive disease of joints and the most common type of arthritis, which is characterized by the degradation of articular cartilage ,107. TheNLRP3 protein expression was increased in patients with knee OA compared with controls in samples from the synovial membrane . FurtherRecent studies have reported that ER stress is associated with OA development ,121. BecGout is an inflammatory arthritis that causes pain and swelling in joints due to monosodium urate (MSU) crystals . Excess NLRP3 polymorphisms and susceptibility to gouty arthritis has been reported [MSU crystals, which enhance IL1β release in synergy with LPS , activatreported . Pseudogreported . CPPD crreported . The majreported ,132,133.Osteoporosis is characterized by a decrease in bone mass, which increases bone fragility and fracture risk . It is cThe levels of inflammatory cytokines are elevated in the absence of estrogen ,141, andProlonged bacterial infection of periodontal tissues induces periodontitis, a condition that affects gums and alveolar bone . The infAmong the proinflammatory cytokines, IL-1 strongly induces bone destruction by promoting osteoclast differentiation and activity ,155,156.Porphyromonas gingivalis, Treponema denticola, and Aggregatibacter actinomycetemcomitans have been implicated in the onset and development of periodontal disease [ disease . Infecti disease ,162,163, disease . Indeed, disease . Moreove disease . Additio disease . Inhibit disease . Activat disease . Taken t disease ,171,172.Aging may be associated with a low level of chronic inflammation, indicated by elevated levels of proinflammatory cytokines . It is kBecause an increase in cortical bone mass has been demonstrated to be a predictor of an enhanced lifespan in mice , attenuaThe NLRP3 inflammasome is involved in the pathogenesis of bone and joint diseases by affecting inflammation, bone resorption, and bone formation. Because the NLRP3 inflammasome and IL-1 signaling pathways are critical factors in the onset and progression of various inflammatory and chronic diseases, blockade of these signaling pathways is an effective therapeutic strategy for the treatment or prevention of NLRP3 inflammasome-related diseases. Indeed, several inhibitors of the IL-1 signaling pathway have already been developed and approved for the treatment of several diseases . For exaIn addition to IL-1 signaling inhibitors, inflammasome inhibitors have been developed. VX-765 is a caspase-1 inhibitor . VX-765


P=0.003). Moreover, ACOT2 exhibited excellent diagnostic efficiency for AML (AUC: 1.000) and related to French-American-British (FAB) classification and cytogenetics. GO, KEGG, and GSEA analyses of 71 common targets between ACOT2 coexpressed and AML-related genes revealed that ACOT2 is closely related to ACOT1, ACOT4, enoyl-acyl carrier protein reductase, mitochondrial (MECR), puromycin-sensitive aminopeptidase (NPEPPS), SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily B member 1 (SMARCB1), and long-chain fatty acid-CoA ligase 1 (ACSL1) in PPI network, and plays a significant role in lipid metabolism, that is, involved in fatty acid elongation and biosynthesis of unsaturated fatty acids. Collectively, the increase of ACOT2 may be an important characteristic of worse OS and abnormal lipid metabolism, suggesting that ACOT2 may become a potential therapeutic target for AML.Acyl-CoA thioesterase (ACOT) plays a considerable role in lipid metabolism, which is closely related to the occurrence and development of cancer, nevertheless, its role has not been fully elucidated in acute myeloid leukemia (AML). To explore the role of ACOT2 in AML and to provide a potential therapeutic target for AML, the expression pattern of ACOT was investigated based on the TNMplot, Gene Expression Profiling Interactive Analysis (GEPIA), and Cancer Cell Line Encyclopedia (CCLE) database, and diagnostic value, prognostic value, and clinical phenotype of ACOT were explored based on data from The Cancer Genome Atlas (TCGA). Functional annotation and enrichment analysis of the common targets between ACOT2 coexpressed and AML-related genes were further performed by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA) analyses. The protein-protein interaction (PPI) network of ACOT2 coexpressed genes and functional ACOT2-related metabolites association network were constructed based on GeneMANIA and Human Metabolome Database. Among ACOTs, ACOT2 was highly expressed in AML compared to normal control subjects according to TNMplot, GEPIA, and CCLE database, which was significantly associated with poor overall survival (OS) in AML ( Acute myeloid leukemia (AML) is a neoplastic disease originating from myeloid progenitor cells characterized by malignant proliferation and differentiation, which is often accompanied by proliferative diseases such as severe leukocytosis –3. AlthoHunt et al. , 10 idenhttps://tnmplot.com/) [The expression level of the type I ACOT family including ACOT1, ACOT2, ACOT4, and ACOT6 were evaluated using the TNMplot platform (ot.com/) , which ihttps://gepia.cancer-pku.cn/) [Gene expression profiling interactive analysis   is a newhttps://www.broadinstitute.org/ccle) [The expression of ACOT1, ACOT2, ACOT4, and ACOT6 originated from type I ACOT family in AML cell lines was verified by the Cancer Cell Line Encyclopedia  , which grg/ccle) . The CCLP < 0.05). The receiver operating characteristic (ROC) curves were generated to evaluate the diagnostic value. The area under the ROC curve (AUC) was used to summarize diagnostic accuracy, with 1.0 representing perfect discrimination and 0.5 representing chance discrimination.The diagnostic and prognostic values of ACOT1, ACOT2, ACOT4, and ACOT6 expression were evaluated in patients with AML based on bone marrow RNA-seq dataset data acquired from TCGA and GTEx databases. The samples were divided into high and low expression groups according to the median expression, and the effects of high and low expression on AML prognosis were analyzed. The overall survival (OS) curve was drawn by a Kaplan–Meier method. The “survival” package (version 3.2-10) was applied for statistical analysis of survival data, and “survminer” package (version 0.4.9) was used for visualization. The log-rank test was applied to evaluate the statistical significance of differences in survival duration  blasts, peripheral blood (PB) blasts, white blood cell (WBC) counts, cytogenetic risks, French-American-British (FAB) classifications, cytogenetics, FLT3 mutation, IDH1 R132 mutation, IDH1 R140 mutation, IDH1 R172 mutation, RAS mutation, and NPM1 mutation was analyzed based on logistic regression models. This analysis was conducted by taking ACOT2 as independent variables, and relevant clinical characteristics were set as dependent variables.https://www.linkedomics.orglogin.php) [https://ctdbase.org/) [https://bioinfogp.cnb.csic.es/tools/venny/index.html).LinkedOmics (gin.php)  is a newse.org/)  was usedhttps://biit.cs.ut.ee/gprofiler/gost) [P < 0.05 was considered statistically significant for GO annotation enrichment analysis and KEGG pathway enrichment analysis.The GO resource provides a platform for functional annotation and enrichment analysis of genes. KEGG is a comprehensive database of biological information designed to assist in the interpretation of large-scale molecular datasets. In the present study, the top 30 core targets of functional protein association network were screened, and GO biological function enrichment analysis and KEGG metabolic process analysis were performed based on g:Profiler (er/gost) . P < 0.0P < 0.05 was considered to be significantly enriched. The Hallmark list from the h.all.v7.2.symbols.GMT was used as the gene set reference.In order to further understand the functional implications of ACOT2, the gene expression files of the 151 cases downloaded from TCGA were separated into high-express groups and low-express groups based on “DESeq2” package (version 1.26.0). In accordance with the differential analysis of high and low expression groups, we sorted the differentially expressed genes (DEG) according to log fold change, then constructed the ordered gene set and enriched it with GSEA based on “clusterProfiler” package (version 3.14.3). https://genemania.org) [GeneMANIA (nia.org)  is a flehttps://hmdb.ca/) [https://www.cytoscape.org) (version 3.8.1). ACOT2-related metabolites were input into Cytoscape software to construct a functional ACOT2-related metabolites association network.The Human Metabolome Database (mdb.ca/)  was usedP < 0.05). The approach directly compared AML and normal samples by grouping all specimens of the same category, and the results were visualized by violin plots. The normal AML comparison was shown in Figures The elevated expression of the type I ACOT  in AML subjects was observed in the TNMplot database  (P=0.091) (P=0.193) (P=0.567)  in AML. One hundred and fifty-one AML samples and corresponding RNA-seq dataset were obtained from TCGA database, however, the information on survival time for 11 samples was missing. Consequently, 140 samples were included in the analysis. The cutoff value of median ACOT1, ACOT2, ACOT4, and ACOT6 expressions were 0.885577317, 2.972574801, 1.491169926, and 0.558179678, respectively. Patients were split into high- and low-expression groups. Results indicated that high expression of ACOT2 was correlated with a significantly poorer prognosis compared with those exhibiting low ACOT2 expression (hazard ratio (HR) = 1.90; P=0.003) , whereasP=0.091) , ACOT4 (P=0.193) , and ACOP=0.567)  was not P=0.567) , ACOT2 (P=0.567) , and ACOP=0.567)  exhibiteP=0.567)  on AML wThe correlation between ACOT2 expression levels and clinical characteristics, including age, gender, patients' race, WBC counts, BM blasts, PB blasts, cytogenetic risks, and FAB classification were analyzed, and the results suggested that ACOT2 did not differ significantly with most of these clinical characteristics in AML except for FAB classification and cytogenetics . CompareNext, the function module of LinkedOmics database was applied to explore the coexpressed genes of ACOT2 in AML. The top 50 significant genes either positively or negatively correlated with ACOT2 were screened out in the heatmap Figures . As showP=0.046) (P=0.046) (P=0.039) (P=0.046)  biological process enrichment analysis of 71 common targets between ACOT2 coexpressed and AML-related genes indicated that the processes were involved in the catalytic activity, myristoyl-CoA hydrolase activity, carboxylic ester hydrolase activity, palmitoyl-CoA hydrolase activity, acyl-CoA hydrolase activity, and CoA hydrolase activity at the MF levels, cytoplasm at the CC levels, and very long-chain fatty acid metabolic process, organonitrogen compound metabolic process, cellular amide metabolic process at the BP levels in AML . The KEGP=0.046) , heme meP=0.046) , interfeP=0.039) , myogeneP=0.046) . These fThe 71 common targets between ACOT2 coexpressed and AML-related genes were uploaded to the GeneMANIA for the protein-protein interactions of common genes analysis . Among tIn the present study, we obtained 53 ACOT2-related metabolites based on Human Metabolome Database . Our further research showed that the expression of ACOT2 was significantly upregulated in most 26 AML cell lines, especially in HNT-34, MOLM-16, BDCM, and THP-1 cell lines. However, there was no significant increase in ACOT1 and ACOT4 in almost any of the AML cell lines, and the expression of ACOT6 in 26 AML cell lines was not available. Survival analysis indicated that the increased level of ACOT2 (P=0.003) was significantly associated with poor OS in AML, whereas ACOT1 (P=0.091), ACOT4 (P=0.193), and ACOT6 (P=0.567) were not significantly correlated. Moreover, the receiver conducting feature curve analysis indicated that ACOT2 (AUC: 1.000) was a prediction of AML and exhibited excellent diagnostic efficiency for AML. Hence, highly expressed ACOT2 is a valuable prognostic factor for AML, indicating that inhibiting the expression of ACOT2 may be a potential approach to prevent the development of AML.So far, metabolic abnormalities have been observed in many tumor diseases, including AML. Nevertheless, there are few cases of specific metabolic enzyme-related gene changes that have been investigated, and the expression and clinical significance of ACOT2 in AML have not been revealed. Here, our research reported that the upregulated expression of ACOT2 and abnormal lipid metabolism were closely related to the clinical prognosis of AML, suggesting that ACOT2-dependent abnormal lipid metabolism may become a prospective target to evaluate the clinical significance of AML diseases. According to the data of TNMplot and GEPIA database, type I ACOT  was highly expressed in AML (all Then, we further studied the correlation between ACOT2 expression and clinical characteristics of AML, and the results suggested that ACOT2 did not differ significantly with most of these clinical characteristics in AML, except for FAB classification and cytogenetics. Compared to the favorable cytogenetic risk group, ACOT2 was highly expressed in intermediate and poor cytogenetic risk groups. Additionally, we examined the association between ACOT2 expression and mutated genes in AML with FLT3, IDH1 R132, IDH1 R140, IDH1 R172, RAS, and NPM1, but no significant correlations were found.Next, the function module of LinkedOmics database was applied to explore the coexpressed genes of ACOT2 in AML, and the top 50 significant genes positively or negatively correlated with ACOT2 were screened out. GO, KEGG, and GSEA analysis of 71 common targets between ACOT2 coexpressed and AML-related genes revealed that the involved processes were the catalytic activity, palmitoyl-CoA hydrolase activity, acyl-CoA hydrolase activity, and CoA hydrolase activity, and very long-chain fatty acid metabolic process, organonitrogen compound metabolic process, cellular amide metabolic process, the most enriched pathways included fatty acid elongation and biosynthesis of unsaturated fatty acids, and the gene sets were significantly enriched in estrogen response early, heme metabolism, interferon alpha response, myogenesis. The PPI network results showed that ACOT2, ACOT1, ACOT4, MECR, NPEPPS, SMARCB1, and ACSL1 played a considerable role in the PPI network, which is related to lipid metabolism. Besides, the ACOT2 and its metabolites interaction network indicated that these metabolites are closely related to lipid metabolism, suggesting that ACOT2 plays significant roles in lipid metabolism involved in fatty acid elongation and biosynthesis of unsaturated fatty acids. Although the metabolites interaction network has greatly expanded the interaction relationships of multiple metabolites, there are still many members whose functions are relatively unexplored and deserve further study.In this study, we explored the mechanism by which high expression of ACOT2 predicts decreased overall survival and abnormal lipid metabolism during AML disease progression. The expression of ACOT2 was significantly upregulated in AML cell lines, which had good diagnostic performance and was significantly associated with poor OS in AML. Besides, ACOT2 plays a significant role in lipid metabolism such as fatty acid elongation and biosynthesis of unsaturated fatty acids. In conclusion, the high expression of ACOT2 may be an important characteristic of decreased overall survival and abnormal lipid metabolism in AML, and the increase of ACOT2 in AML can be used as a potential biomarker for auxiliary diagnosis and prognosis, and targeted ACOT2 may be a promising approach to inhibit AML progression.


Nitrogen (N) is a vital mineral component that can restrict the growth and development of plants if supplied inappropriately. In order to benefit their growth and development, plants have complex physiological and structural responses to changes in their nitrogen supply. As higher plants have multiple organs with varying functions and nutritional requirements, they coordinate their responses at the whole-plant level based on local and long-distance signaling pathways. It has been suggested that phytohormones are signaling substances in such pathways. The nitrogen signaling pathway is closely associated with phytohormones such as auxin (AUX), abscisic acid (ABA), cytokinins (CKs), ethylene (ETH), brassinosteroid (BR), strigolactones (SLs), jasmonic acid (JA), and salicylic acid (SA). Recent research has shed light on how nitrogen and phytohormones interact to modulate physiology and morphology. This review provides a summary of the research on how phytohormone signaling affects root system architecture (RSA) in response to nitrogen availability. Overall, this review contributes to identifying recent developments in the interaction between phytohormones and N, as well as serving as a foundation for further study. Enhancing crop yields and reducing environmental risks simultaneously is a massive challenge in sustainable agricultural development. In the past four decades, agricultural food production has doubled worldwide, and nitrogen (N) fertilizer use has increased seven-fold. Plants require N as a major macronutrient, and its availability has been recognized for years as a critical factor in crop production and food security ,2. The oEnhancing NUE in plants is crucial to enhancing yields and quality, reducing nutrient input costs, and improving soil, water, and air quality . In addi3−) or ammonium (NH4+), or organically, mainly as free amino acids ..MedicagoLATD may regulate the activation of the meristems through a nitrate-ABA signaling pathway due to its homology to nitrate transporters , su, su3− avA. thaliana and rice [DR5:: GUS reporting method and reverse genetic approaches [Arabidopsis in response to nitrate [Auxins are a group of important phytohormones that modulate plant development and morphology to environmental conditions and are vital under nitrogen stress ,104,105.and rice ,107. Auxproaches ,49. Auxi nitrate ,109,110. nitrate ,112. As  nitrate ,114. Mor nitrate ,116,117.A. thaliana. Furthermore, plants grown in a medium with LN levels produced lower levels of auxins in their shoots than in a medium with HN [Arabidopsis seedlings cultivated under LN conditions produced greater root auxin levels than those grown under HN conditions, which indicates that dicot and monocot plants share a similar mechanism for controlling root auxin levels according to the nitrogen level of the plant [Similarly, a switch from a high-nitrate to a low-nitrate medium showed that auxin, in the roots, increased following lateral root growth in  with HN ,119. Addhe plant . Howeverhe plant .A recent study in understanding auxin action suggests that the driving force of auxin-regulated growth and development is the concentration gradient and the differential sensitivity of various cell types, apart from the auxin levels in tissue. Cell-to-cell polar transport establishes the auxin gradient, and the differential sensitivity is achieved by modulating signaling components ,122. RecArabidopsis [Arabidopsis have been identified, including TAR2, a gene involved in auxin biosynthesis, auxin transporters such as PIN1, PIN2, PIN4, and PIN7, as well as molecular components that control their subcellular distribution [AUXIN RESPONSE FACTOR 8 (ARF8) was identified as involved in auxin signaling  trafficking and partitioning along the primary root and root growth [IAA production by triggering tryptophan aminotransferase (TAA1) and tryptophan aminotransferase-related (TAR1 and TAR2) enzymes, which have similar roles in ethylene responses [Ethylene is a master regulator of root growth and development due to its involvement in both t growth . Ethylenesponses . It has esponses .NRT) genes [3−/ethylene interactions [AMINOCYCLOPROPANE CARBOXYLIC ACID SYNTHASE (ACC) and AMINOETHOXYVINYLGLYCINE (AVG) at low and high nitrate concentrations led to the upregulation and downregulation of nitrate transporters (AtNRT1.1 and AtNRT2.1) [etr1-3 and ein2-1 mutants showed no sensitivity to high nitrate concentrations [AtNRT genes  ethylene response to changes in external nitrate availability through the expression of nitrate transporter (T) genes ,147. Seeractions . Anotherractions . FollowitNRT2.1) . Howevertrations , concludRT genes A. AtNRT2.1 rather than AtNRT1.1 was associated with ethylene biosynthesis and signaling responses in seedlings transferred from high to low nitrate concentrations [AtNRT2.1 in wild-type and mutant forms of the ctr1-1, ein3-1, and eil-1 genes revealed that ethylene repressed AtNRT2.1 and nitrate acquisition through one of the ethylene signaling cascade’s components [AtNRT2.1 expression  or sufficient (10 mM NO3−).An interesting study reveals that ethylene may serve as a signal for plant-to-plant communication in rice under high-density stress conditions when ethylene production is linked to genes involved in ethylene homeostasis . ResearcBrassica juncea plants, N availability impacts photosynthesis, stomatal conductance, and growth and influences ethylene’s evolution [ethephon  improved ethylene production and photosynthesis of plants grown with N levels of 40 and 80 mg kg−1 [In volution ,153. Acc mg kg−1 . Table 4Arabidopsis, nitrogen deficiency promotes BR biosynthesis by upregulating DWF1, CPD, DWF4, and BR6ox2 [CPD, DWF4, and DWF1 in maize and rapeseed, respectively [An increasing body of research shows that phosphate (Pi) or boron (B) deficiency suppresses brassinosteroid (BR) levels, whereas N deprivation elevates them ,159. In d BR6ox2 ,161. Morectively ,163, impMAX3 and MAX4 are consistently increased by N and Pi deficiency in A. thaliana, similar to Oryza sativa D17 and D10 [Many plant species produce strigolactones as plant hormones . Despite and D10 ,168.jasmonic acid (JA) and jasmonic acid–isoleucine (JA-Ile) in A. thaliana roots [Low levels of N have been demonstrated to trigger an undisclosed mechanism that reduces the concentrations of na roots ,170,171.There is currently limited understanding of how the availability of nutrients affects the production of salicylic acid (SA) in the roots. Two recent studies have demonstrated that roots that lack both N and Pi accumulate more SA ,173. DesCK, ABA, AUX, ETH, BR, SL, JA, and SA) are the key players that regulate several signaling pathways in response to nitrogen stress and modulate root system architecture  that regulates RSA [This review comprehensively explains the molecular mechanisms of phytohormones in nitrogen signaling. Phytohormones  and breeding crops with high yields using low nitrogen fertilizer inputs will be essential in the future with the use of advanced biotechnologies and accumulated basic research findings.