Head and Neck Squamous Cell Carcinomas (HNSCCs) are cancers that originate from cells of the head and neck region, including the mouth, nose, and throat. The diversity of these cell types is also mirrored by the high number of different mutations that promote cancer development and progression. For the clinical management of this disease, it is important to identify biomarkers that allow early detection or predict relapse and resistance to therapy. A non-invasive way to monitor these markers over time are so called liquid biopsies, which mostly refers to the detection and analysis of tumor cells or cell-free DNA (cfDNA) in the blood of patients. This review summarizes our current understanding of HNSCC genetics and discusses how the detection of genetic variation in the cfDNA of HNSCC patients can be used to monitor disease and guide therapy.Tumors shed cell-free DNA (cfDNA) into the plasma. “Liquid biopsies” are a diagnostic test to analyze cfDNA in order to detect minimal residual cancer, profile the genomic tumor landscape, and monitor cancers non-invasively over time. This technique may be useful in patients with head and neck squamous cell carcinoma (HNSCC) due to genetic tumor heterogeneity and limitations in imaging sensitivity. However, there are technical challenges that need to be overcome for the widespread use of liquid biopsy in the clinical management of these patients. In this review, we discuss our current understanding of HNSCC genetics and the role of cfDNA genomic analyses as an emerging precision diagnostic tool. Head and neck squamous cell carcinoma (HNSCC) is a type of cancer that affects the epithelial cells that line regions of the head and neck, including the mouth, nose, and throat  1,2,3,4,3,42,3,4The identification of non-invasive, blood or saliva-based biomarkers to detect HNSCC relapse would be a significant advance in the management of this disease. Cell-free deoxyribonucleic acid (cfDNA), is a promising candidate for such a biomarker, as it has been shown to reflect the presence of residual tumor cells and may be able to predict imminent relapse in different solid tumor settings ,15. The In this review, we discuss our current understanding of HNSCC genetics and the potential role of cfDNA genomic analyses as an emerging precision diagnostic tool in the management of these patients.TP63, SOX2 and PIK3CA, or amplification of 11q13 regions encompassing FADD and PPFIA1), distinct CNV patterns reflect the HPV-associated dichotomization of HNSCC entities (E2F1), 12p13  and 3q27/28 , while HPV- HNSCC cells are enriched for amplifications of 7p11 (EGFR), 8p11 (FGFR1), 17q12 , 5p15 (TERT), 9p24 , 15q26  and the coamplification of 11q13 (CCND1) and 11q22  [BIRC2, BIRC3, ATM), 14q32.32 (TRAF3), 13q14.2  or 7q36.1 , while HPV- HNSCCs are commonly deleted at 9p21.3 (CDKN2A), 2q22.1 (LRP1B), 10q23.31 (PTEN), 9q34.3 (NOTCH1) and 18q21.2 (SMAD4) [Despite originating from a variety of different tissues in the upper aerodigestive tract, HNSCC cells display recurrent patterns of structural genome aberrations and acquired somatic mutations ,30,31,32entities  [20,22,22, YAP1) ,22,23,24 (SMAD4) . NPCs shTP53, FAT1, CDKN2A, PIK3CA, NOTCH1, KMT2D, NSD1, CASP8 and AJUBA genes [TP53 with frequencies ranging up to 80% of HPV- cases [TP53 mutations , they uBV- NPCs . The TP5BV- NPCs ,44,45,46BV- NPCs ,47. FollBV- NPCs ,44,47. ABV- NPCs ,49. In HBV- NPCs , promoteBV- NPCs  and genoBV- NPCs , result BV- NPCs . As a cosurvival .TP53, the second most mutated gene in HNSCC is FAT1, affecting around 20% of patients with HNSCC [FAT1 encodes a multifunctional type 1 transmembrane cadherin-related protein that—after proteolytic activation—acts as a modulator of oncogenic Wnt/β-catenin signaling [After th HNSCC ,57. FAT1ignaling , the Hipignaling , Ena/VASignaling  and the ignaling . FAT1 muignaling .CDKN2A, NOTCH1, and PIK3CA are found in about 10–30% of HNSCC cases dependent on cohort size and patient selection [CDKN2A encodes the p16INK4A tumor suppressor which arrests cell cycle progression at the G1-S restriction point by inhibiting CDK4/6-mediated phosphorylation of retinoblastoma-associated protein (RB1), a prototypical cell cycle regulator and driver of carcinogenesis [CDKN2A loss is associated with resistance to immunotherapy [INK4A. In HPV- HNSCCs, TP53 loss-of-function mutations are associated with CDKN2A point mutations or the loss of 9p21.3, which encodes CDKN2A, while in HPV+ HNSCCs, TP53 and RB1 are degraded by the viral E6 and E7 oncoproteins [NOTCH1 tumor suppressor gene encodes a conserved transmembrane protein that exerts pivotal regulatory roles during development and substantially contributes to tissue homeostasis [NOTCH [TP53 [NOTCH, CDKN2A and the action of the TP53 family member TP63 [PIK3CA [Mutations in election ,38,62,63ogenesis . CDKN2A otherapy . Interesproteins ,25,38. Teostasis ,67. Alths [NOTCH  and TP53CH [TP53 , it is aCH [TP53 . Epithelber TP63 ,71,72. Iber TP63 . The detber TP63 . Notably [PIK3CA ,73,74. AALK, ROS or RET are rarely detected in HNSCC [ETV6-NTRK3 [PAN3-NTRK2 [FGFR3-TACC3 [In addition, classical tumor-driving gene fusions of in HNSCC . NeverthV6-NTRK3 , PAN3-NTN3-NTRK2  and FGFRR3-TACC3  have beeHRAS activating mutations, high microsatellite instability (MSI-H), high tumor mutational burden (TMB-high), NTRK fusions, CDKN2A inactivating alterations, and EGFR amplifications [Patients with HNSCC, who have failed standard first-line therapies, have limited therapeutic options and may benefit from new targeted therapies. Marret et al. ranked recurrent molecular alterations in HNSCC on the basis of the European Society for Medical Oncology (ESMO) Scale for Clinical Actionability of Molecular Targets (ESCAT) and identified six of 33 actionable alterations as the most clinically relevant: ications .HRAS mutant (mHRAS) HNSCC xenograft models [HRAS 20 patients were evaluable for response at the time of data cutoff [HRAS variant allele frequency (VAF) of ≥20% was 55% , and median progression-free survival (PFS) on tipifarnib was 5.6 months  versus 3.6 months  on the last prior therapy, and the median overall survival (OS) was 15.4 months  [HRAS HNSCC with VAF ≥ 20% after disease progression on platinum-based chemotherapy in 2021.HRAS-activating mutations occur in approximately 4–8% of HNSCC patients . HRAS ont models . In a sia cutoff . The objto 29.7) . Due to In HNSCC the incidence of TMB-high, defined as ≥10 mutations per megabase (mut/Mb), is around 20% and the incidence of MSI-H is 1.2% . TMB-higNTRK fusions are rare in HNSCC (<1%) [NTRK gene fusions: the tropomyosin kinase (TRK) inhibitors entrectinib and larotrectinib [NTRK fusion-positive solid tumors, the objective response rates ranged from 57% to 79% resulting in tissue-agnostic approvals by the EMA and FDA [CC (<1%) . There arectinib ,84. In p and FDA ,83,84.CDKN2A inactivating alterations that cause the hyperactivation of CDK4/6 are reported in 53.8% of HNSCC [CDKN2A mutations [of HNSCC . The selof HNSCC . Howeverof HNSCC . Phase 2utations .EGFR amplifications are commonly found in patients with HNSCC [EGFR-amplified tumors [th HNSCC . Afatinith HNSCC . Compareth HNSCC . A more d tumors .EGFR copy number was not a predictive biomarker for the efficacy of cetuximab [521 represents an important mechanism of primary resistance to cetuximab in HNSCC [TP53 is the most frequently altered gene in HNSCC with mutations detected in over two-thirds of patients [TP53 mutational status may, however, predict decreased sensitivity to cisplatin-based therapy [In combination with chemotherapy in the first-line treatment of R/M HNSCC, etuximab . Howeverin HNSCC . This EGin HNSCC . TP53 ispatients  but evidpatients . TP53 mu therapy . Loss of therapy .The spatiotemporal genetic heterogeneity of solid tumors has been associated with a dismal prognosis due to decreased therapy response and higher rates of tumor recurrence ,96. ThisEfforts to break the heterogeneous group of HNSCCs down into prognostically relevant subgroups utilized the bulk gene expression analysis of more than 279 tumor samples to define four distinct tumor expression subtypes ,23,100. Another study has shed light on the genetic patterns of metastasis in HNSCCs, revealing two distinct subtypes with potential clinical implications. Patients with hematogenous metastasis exhibited upregulations of PD-L1 and PD-L2, suggesting that immune checkpoint inhibition may be a viable treatment option for this group. In contrast, patients with lymphatic metastasis showed a better response to chemotherapy in combination with locoregional radiotherapy .In addition, prognostically relevant tumorigenic mutations were also found in tumor-adjacent tissue sometimes referred to as oral field cancerization ,69,102. Taken together, liquid biopsy methods can indeed help to further elucidate the mutational landscapes of tumors, their surrounding tissue as well as their metastases. However, this complex spatiotemporal heterogeneity poses a great challenge and needs to be factored in for the future clinical application of liquid biopsy. Different techniques may need to be applied.The analysis of cfDNA already generated multiple insights into tumor genetic composition , resistaThe genetic heterogeneity of the primary tumor and metastatic lesions, tumor evolution on therapy or surveillance, shared genetic mutations of precursor lesions and coexisting germline mutations or clonal hematopoiesis are relevant biological challenges for all cfDNA analysis techniques . AdditioIn addition to biological obstacles, the technical limitations of cfDNA analysis present significant challenges. During the pre-analytical phase, the use of specialized collection tubes with reagents for leukocyte stabilization can allow for the extension of storage and shipping times by up to 14 days at room temperature. In comparison, clinical practice often uses EDTA tubes which only offer a window of 2–4 h for further downstream processing ,116. GenObtaining an accurate estimate of input cfDNA and minimizing gDNA contamination is crucial for precise estimations of potential tumor gene amplifications, such as HER2, which have important treatment implications. Typically, gene amplifications are deduced by calculating the relative ratio of the target gene to a copy number reference gene in close chromosomal proximity, which helps to exclude potential biases caused by genomic aneuploidy ,120. In Different techniques for cfDNA analysis have been developed, of which variants of next-generation sequencing, either amplicon- or capture-based . To ensure the robustness and standardization of these assays quality standards and controls are proposed which will help with the translation into clinical practice .In HNSCC, studies examining cfDNA have not only focused on blood but also on saliva samples ,140. DNAn = 71) and before salvage treatment for recurrent HNSCC (n = 22) [To explore the utility of tumor-derived DNA from different body sites for the diagnosis and surveillance of HNSCC, Wang et al. collected saliva and plasma before definite treatment for primary HNSCC ((n = 22) . Each tu(n = 22) . In sali(n = 22) . In plas(n = 22) . Thus, t(n = 22) . Overall(n = 22) . Further(n = 22) .PAX5, Endothelin Receptor β (EDNRB), and Deleted in Colorectal Cancer (DCC) was studied in HNSCC biopsies and corresponding pretreatment oral rinses [PAX5 with a sensitivity in oral rinses of 84.0%  and a specificity of 87.9%  [Tumor-specific alterations such as gene methylation represent a strategy to differentiate between tumor-free circulating DNA and tumor-derived cfDNA in HNSCC patients . Fung etl rinses . The besto 95.0) ; 76.9% oto 95.0) .PAX5 methylation analyzed by ddPCR technology was also used to assess histologically cancer-negative deep surgical margin samples obtained from 82 HNSCC surgeries by an imprinting procedure and primary tissue collection [PAX5 methylated imprint margins were an excellent predictor of poor locoregional recurrence-free survival  by multivariate analysis [Interestingly, llection . PAX5 meanalysis .Notably, the association of HNSCC with HPV or EBV infection offers the potential to use virus-derived cfDNA as a marker. For example, it has been reported that circulating HPV DNA correlates with tumor burden  or stagiA few studies have investigated minimal residual disease (MRD) detection by mutant cfDNA analysis in patients with HNSCC who underwent resection with curative intent ,148,149.TP53 mutations in blood samples from HNSCC patients with the locoregional disease is generally feasible, opening up avenues for post-treatment surveillance [n = 11) had a significantly better prognosis than those who became cfDNA positive [Van Ginkel et al. show that ddPCR-based detection of eillance . Jonas eeillance . Moreoveeillance . With theillance . Two of eillance . Longitueillance . In seveeillance . Patientpositive .Despite the low patient numbers included in these trials, these studies collectively indicate that patients with post-surgical detection of mutant cfDNA eventually relapse. Liquid biopsy positivity typically precedes clinical relapse by several months.Liquid biopsies have the potential to enhance precision medicine for patients with advanced HNSCC. However, only a limited number of studies have shown the effectiveness of this method for detecting druggable lesions and monitoring disease and resistance in patients with R/M HNSCC.TP53 alterations and HPV-negative status was associated with increased benefit from the combination, indicating that phosphatidylinositol 3-kinase (PI3K) inhibition may improve outcomes in this subset of patients historically characterized by poorer clinical outcome [NOTCH1 mutation in the tissue sample were included in this trial based on preclinical data supporting the susceptibility to this drug in this subset of patients. The cfDNA-based detection of NOTCH1 mutations showed satisfactory concordance with tissue analysis, suggesting that this biomarker can be conveniently detected in the blood for future trials. However, to our knowledge, there are no data from HNSCC trials available with biomarker-guided patient selection based on cfDNA analysis. In this trial, changes in the cfDNA quantity during treatment were consistent with the clinical course and cfDNA collected at the time of disease progression showed new molecular alterations such as PIK3CA, BRAF, and TP53 mutations [Galot et al. determined the utility of liquid biopsy to detect potentially actionable mutations in cfDNA . They fo outcome . Interes outcome ,153. Ano outcome . Only pautations .EGFR, KRAS, NRAS, and HRAS. In patients with on-treatment progression, 46% showed acquired RAS mutations in cfDNA before clinical resistance emerged, indicating a significant correlation between the emergence of RAS mutant clones and clinical resistance. The study also showed the potential of liquid biopsies to detect imminent resistance before clinical progression occurs.In addition to pre-therapeutic screening in patients with R/M HNSCC, serial analysis of cfDNA may provide insights into tumor control and the development of resistance traits over time. Our own study investigated tumor evolution in patients with R/M HNSCC treated with cetuximab/platinum/5-fluorouracil . The stuThe ongoing FOCUS study (NCT05075122) investigates the combination of a cancer vaccine with pembrolizumab in patients with R/M HNSCC. The biomarker part of this study includes serial liquid biopsy monitoring in R/M HNSCC as a predictor of disease progression. To search for potentially emerging resistant tumor subclones, the liquid biopsy panel contains genes previously reported to be involved in resistance to checkpoint inhibitors.For HNSCC, there is a great need for the identification of new biomarkers due to the high risk of relapse in locoregional disease after initial treatment and the limited therapeutic options in the metastatic setting. These biomarkers should identify minimal residual disease, assess treatment response, monitor disease activity, profile genomic tumor landscape, and detect targetable alterations and resistance mechanisms. All this can be offered by the analysis of cfDNA as a new type of specific and non-invasive biomarker, detected through liquid biopsy. In HNSCC, the analysis can be performed in both the blood and saliva with increased sensitivity when both analyses are combined. However, the limitations of this method should be taken into consideration. One crucial factor in analyzing cfDNA is the technical procedure as well as the standardization of the assays. The currently available data for HNSCC show that the potential usefulness of a distinct platform and/or marker depends on the respective cancer entity, stage and also diagnostic aim. More clinical studies will be necessary to investigate whether a change in therapy, based on this cfDNA analysis, will improve patient outcomes as the primary endpoint, compared to the current standard procedure consisting of clinical and radiological measures. These studies also need to define appropriate genetic markers for specific clinical endpoints, since liquid biopsy approaches remain purely experimental at this stage. Only when these data are available, will liquid biopsy fulfill its promise as a cost-effective, minimally invasive approach for cancer diagnostics.To conclude, cfDNA analysis is becoming increasingly important in the management of HNSCC patients in the context of personalized and precision cancer medicine. Nevertheless, despite encouraging data, further research is mandatory in order to shed more light on this analysis before it is widely integrated into daily clinical practice.


MDH genes in sweet potato (Ipomoea batatas) and its two diploid wild relatives, Ipomoea trifida and Ipomoea triloba, respectively, were identified. These MDH genes were unevenly distributed on seven different chromosomes among the three species. The gene duplications and nucleotide substitution analysis (Ka/Ks) revealed that the MDH genes went through segmental duplications during their evolution under purifying selection. A phylogenetic and conserved structure divided these MDH genes into five subgroups. An expression analysis indicated that the MDH genes were omni-presently expressed in distinct tissues and responded to various abiotic stresses. A transcription factor prediction analysis proved that Dof, MADS-box, and MYB were the main transcription factors of sweet potato MDH genes. These findings provide molecular features of the MDH family in sweet potato and its two diploid wild relatives, which further supports functional characterizations.Malate dehydrogenase  plays a vital role in plant growth and development as well as abiotic stress responses, and it is widely present in plants. However, the MDH family genes have not been explored in sweet potato. In this study, nine, ten, and ten  As it is an oxidoreductase ubiquitous in plants, animals, and microorganisms, malate dehydrogenase (MDH) catalyzes the reversible conversion of malate to oxaloacetate (OAA) with nicotinamide adenine dinucleotide (NAD+) or nicotinamide adenine dinucleotide phosphate (NADP+) as a coenzyme factor . NAD-depArabidopsis [Arabidopsis, NAD-MDHs and cMDHs have been found to play important roles in embryonic development [MDHs are involved in abiotic stress responses. Under a salinity treatment, the overexpression of SbNADP-ME increases the Arabidopsis germination rate and root length, which improves the seedling salt tolerance [ZmNADP-MDH improves photosynthesis and salt tolerance in Arabidopsis [MdcyMDH overexpression enhances the tolerance to cold and salt stresses by producing more reductive redox states and increasing the salicylic acid level [OsMDH12 rice is susceptible to salt stress, but the OsMDH12 mutant exhibits enhanced salt endurance [MDH genes responding to salt stress are also found in tomato [MDH genes participate in regulating the nutrient deficiency response. Chinese fir 8 ClMDH genes are upregulated under low-phosphorus stress [GhmMDH1-overexpressing plants produce significantly longer roots and have higher biomasses and phosphorus contents than WT plants under phosphorus deficiency [MDH genes are involved in aluminum stress response; for example, the overexpression of malate dehydrogenase in alfalfa [So far, MDHs have been identified in various plants, including 9 MDHs in bidopsis ,12, 12 ibidopsis , 13 in cbidopsis , 16 in sbidopsis , 12 in tbidopsis , 12 in abidopsis , 16 in pbidopsis , and 12 bidopsis . In Arabelopment ,22, leafelopment , energy elopment , and theelopment ,26,27. Solerance . The ovebidopsis . In applid level ,31. Overndurance . MDH genn tomato . In addis stress . The cotficiency . MDH gen alfalfa  and toba alfalfa  confers I. batatas (L.) Lam) is also an important energy crop for its high edible, feed, and medicinal values [MDH gene families has been carried out in various plants, but there has not been a report about sweet potato MDH genes. In recent years, highly assembled genomes of sweet potato hexaploid [I. trifida (2n = 2x = 30) and I. triloba (2n = 2x = 30) [MDH gene of sweet potato and its two diploid relatives to reveal their additional biological functions.Being the seventh largest food crop in the world, sweet potato  , have beMDH genes from sweet potato and its two diploid wild relatives were identified with high confidence. A collinear analysis showed that segmental duplication was the main driver of the evolution of sweet potatoes and their relatives. An evolutionary analysis was performed for these MDH genes. Moreover, an expression profiling analysis revealed that these MDH genes were expressed in distinct tissues and actively responded to distinct environmental stresses, such as salinity, drought, and low temperatures. Overall, our findings provide a theoretical basis to further investigate the functions of sweet potato MDH genes.In this study, 29 MDHs were identified in I. batatas, I. trifida, and I. triloba, respectively, and they were named IbMDH 1-9, ItfMDH 1-10, and ItbMDH 1-10 according to their locations on the chromosomes. In sweet potato, the CDS length of IbMDHs ranged from 981 bp (IbMDH2) to 1326 bp (IbMDH9), and the genomic length varied from 2040 bp to 4842 bp. The lengths of the protein sequences ranged from 326 aa (IbMDH2) to 441 aa (IbMDH9), with a molecular weight (MW) of 34.59 kDa to 48.18 kDa. The isoelectric points (pI) ranged from 6.10 (IbMDH2) to 8.74 (IbMDH7). Most IbMDHs were stable with an instability index of less than 40. The GRAVY values of IbMDHs varied from −0.182 to 0.211. A subcellular localization prediction showed that most IbMDHs were in the chloroplast, IbMDH1, and IbMAH3 in the cytoplasm, and IbMDH7 in the mitochondria   , and the51.1 Mya . MDH gene family among I. batatas, I. trifida, I. triloba, Arabidopsis thaliana, Oryza sativa, and Gossypium arboreum, a phylogenetic tree for 63 MDHs of these six species  was constructed. The results showed that all of the MDHs were divided into five groups : Group 1 , Group 2 , Group 3 , Group 4 , and Group 5   . These rMDHs contained these ten conserved motifs but varied within different subgroups. The MDH genes in Group 1 missed five motifs ; in Group 2, the MDHs missed motif 3, motif 4, and motif 6, where IbMDH9 in Group 2 also missed motif 8; in Group 3, the MDHs missed motif 9; in Group 4 and Group 5, the MDHs lacked motif 9, where IbMDH5 in Group 4 missed motif 6 and IbMDH3 in Group 5 missed motif 4. Therefore, the MDH genes showed some differences throughout their evolutions.The distribution of the conserved motifs prediction using the MEME website found that ten conserved motifs were identified containing 15 to 50 amino acids . Most ofMDHs showed some variations  were higher in the primary roots than in other tissues. Additionally, the tissue-specific expression pattern was observed. For instance, the IbMDH4 and IbMDH8 genes were expressed only in the leaves, and the IbMDH9 transcript was abundant in the leaves and stems. The transcript levels of these ing qPCR a. The reMDHs in six tissues  of I. trifida and I. triloba were analyzed . Under a low-potassium treatment, except for IbMDH1, which was upregulated in “Xushu 32” and downregulated in “Ningzishu 1”, the transcript levels of eight sweet potato MDH genes were significantly upregulated in both varieties   . In fibregulated .ItfMDHs and ItbMDHs were also analyzed using the RNA-seq data of I. trifida and I. triloba under the ABA, GA3, and IAA treatments [ItfMDHs  were strongly induced, and only ItfMDH8 was downregulated, and the others did not show significant changes. Under the GA3 treatment, ItfMDH4 and ItfMDH7 were upregulated. Under the IAA treatment, ItfMDH3 was upregulated. In I. triloba, under the ABA treatment, ItbMDH4 and ItbMDH8 were upregulated, and the others were downregulated. Under the GA3 treatment, ItbMDH3 was upregulated, and ItbMDH9 was downregulated. Under the IAA treatment, ItbMDH7 was upregulated, and ItbMDH1, ItbMDH4, and ItbMDH10 were downregulated  were upregulated upon both stresses, showing similar expression patterns  were upregulated after both stresses, and IbMDH4, IbMDH7, and IbMDH8 were upregulated only under drought stress  , IbMDH9 (34), and IbMDH2 (32), while IbMDH3 was targeted least, by only 14 TFs  . Among ty 14 TFs .I. trifida and I. triloba, have often been studied [MDH genes were identified in I. batatas, I. trifida, and I. triloba, respectively, which is less than rice [MDH genes were scattered on seven chromosomes, several of which contained two MDH genes, such as LG13 and LG15 in sweet potatoes, and Chr02 and Chr06 in I. trifida and I. triloba , which could lead to functional differences. Based on the phylogenetic tree, the MDH gene family was divided into five subgroups  of the sweet potato MDH genes showed high expression patterns in the primary roots, and some MDH genes exhibited tissue-specific expressions; for instance, IbMDH4 and IbMDH8 were expressed only in the leaves, and IbMDH9 was expressed in the leaves and stems  ItfMDHs and 50% (5/10) ItbMDHs were highly expressed in the flower buds, and other MDH genes were also expressed in the roots, stems, and leaves  and ABRE (ABA-responsive element) were abundant in the MDH gene promoters  of the IbMDHs in the stems and 56% (5/9) in the leaves were upregulated under both stresses. Notably, sweet potato MDH transcripts responded more to drought stress than salt stress. For example, the transcript levels of IbMDH1 and IbMDH3 were increased by 168-fold and 12-fold upon drought stress, but increased by 55-fold and 7-fold under salt stress, and IbMDH4, IbMDH7, and IbMDH8 in the leaves were upregulated only after drought stress  and Ipomoea Genome Hub . The Arabidopsis MDH proteins were downloaded from the TAIR database , and Oryza sativa and Gossypium arboreum MDH proteins were from Phytozome v13  [MDH genes, two methods were utilized. First, HMMsearch (ver. 3.1b2) with default settings was used to search the protein sequences of the three species for the MDH domain (Pfam accession numbers: PF00056 and PF02866) [Arabidopsis MDH protein sequences were used as queries in a BLASTP (ver. 2.10.0+) search against the protein database of each species with an E-value threshold of 1 × 10–10 [https://www.ncbi.nlm.nih.gov/Structure/bwrpsb/bwrpsb.cgi, accessed on 19 July 2023) [http://smart.embl-heidelberg.de/, accessed on 19 July 2023) [http://web.expasy.org/protparam/, accessed on 20 July 2023) was used to calculate protein physicochemical parameters [https://wolfpsort.hgc.jp/, accessed on 20 July 2023) was used to create subcellular localization predictions [The protein sequences and GFF gene annotation files of sweet potato, ly 2023) . To idenPF02866) . Second, × 10–10 . Then, aly 2023)  and the ly 2023) . Finallyrameters . WoLF PSdictions .IbMDHs, ItfMDHs, and ItbMDHs on the chromosomes. The collinearity relationships of MDH genes within species were constructed using MCScanX [TBtools software (v.2.001)  was used MCScanX  and visuMDHs among the whole genomes of I. batatas, I. trifida, and I. triloba was performed using MCScanX [MDH genes were estimated using the TBtools software’s Ka/Ks calculation package. The estimated time of detergence  was calculated as described previously [The synteny analysis of  MCScanX  and visueviously .I. batatas, I. trifida, I. triloba, Arabidopsis, O. sativa, and G. arboreum was performed using ClustalW in MEGA 7 [http://www.evolgenius.info/evolview/, accessed on 28 August 2023) [The phylogenetic analysis of MDHs from n MEGA 7  with defst 2023)  was usedhttps://meme-suite.org/meme/, accessed on 28 August 2023) analyzed conserved motifs of MDH proteins, and the maximum number of motifs parameter was set to 10 [The MEME tool  website [MDH genes. Then, the TBtools software was used to visualize the cis-regulatory element figure. The upstream promoter region (2000 bp) of the  website , which ihttps://string-db.org/, accessed on 12 September 2023) was utilized to predict and execute potential protein–protein interaction networks using sweet potato MDH proteins based on known Arabidopsis homologs. Using the default parameters, the online STRING database 23  was usedsion 3.8 .MDH gene expression profile analysis. Three bio project datasets  were downloaded from the NCBI database. Another one was our in-house (unpublished) sweet potato heat treatment. Among them, “Xushu 18” was used for hormonal treatment, cold-tolerant “Liaohanshu 21” and cold-sensitive “Shenshu 28” were used for cold treatment, low-k-tolerant “Xushu 32” and low-k-sensitive “Ningzishu 1” were usedfor low-potassium treatment, and heat-tolerant “Guangshu 87” and heat-sensitive “Ziluolan” were used for heat treatment. Additionally, the gene expression data in I. trifida and I. triloba were downloaded from the Sweet Potato Genomics Resource . The MDH expression was measured in fragments per kilobase of exon per million fragments mapped (FPKM) [Four transcriptome bio project datasets were chosen for the sweet potato d (FPKM) . The heaI. batatas) cultivar “Jishu 26” was used for qRT-PCR analysis in this study. Sweet potato plants were cultivated in a field at the experimental field of Guangdong Ocean University, Guangdong, China. For tissue expression, the flower, leaf, stem, primary root, firewood root, and tuberous root tissues were sampled from 3-month-old “Jishu 26” planted in the field. For the abiotic stress treatments, twigs about 30 cm in length from 3-month-old field-grown “Jishu 26” were cultured in the Hoagland solution for 14 days to treat them: for salt stress treatment, the twigs were cultured in the Hoagland solution with 0 and 200 mM NaCl, and for drought stress treatments, the twigs were cultured in Hoagland solution with 0 and 300 mM mannitol. The primary root, stem, and leaf samples were collected at 0, 8, 16, and 24 h after the treatments. The sweet potato . The qRT-PCR experiments were carried out based on previous studies . Three beference .MDH genes were identified in sweet potato and its two diploid relatives. A conserved motif and gene structure analysis showed the conservation and dispersion of MDHs during evolution. The expression profiles indicated that the MDHs showed tissue specificity and various expression patterns in sweet potato and its two diploid relatives. MDH genes may be involved in various hormone and abiotic stress responses to regulate plant growth and development. The findings in this study will help us better understand the biological functions of MDHs in coping with climate change and identify possible candidate genes for enhancing field and abiotic stress tolerance in sweet potato and its two diploid relatives.In this study, 29


MLL) gene rearrangements, and a subset of de novo acute myeloid leukemia. Recent research has shown a sustained interest in determining the regulators of cytochrome P450, family 2, subfamily E, member 1 (CYP2E1) expression and activity as an emerging field that requires further investigation in acute myeloid leukemia evolution. Therefore, this review suggests that CYP2E1 and its mutations can be a therapeutic or diagnostic target in acute myeloid leukemia.Oxidative stress is associated with several acute and chronic disorders, including hematological malignancies such as acute myeloid leukemia, the most prevalent acute leukemia in adults. Xenobiotics are usually harmless compounds that may be detrimental, such as pharmaceuticals, environmental pollutants, cosmetics, and even food additives. The storage of xenobiotics can serve as a defense mechanism or a means of bioaccumulation, leading to adverse effects. During the absorption, metabolism, and cellular excretion of xenobiotics, three steps may be distinguished: (i) inflow by transporter enzymes, (ii) phases I and II, and (iii) phase III. Phase I enzymes, such as those in the cytochrome P450 superfamily, catalyze the conversion of xenobiotics into more polar compounds, contributing to an elevated acute myeloid leukemia risk. Furthermore, genetic polymorphism influences the variability and susceptibility of related myeloid neoplasms, infant leukemias associated with mixed-lineage leukemia ( Acute myeloid leukemia is a cancerous condition that affects hemopoietic stem cells or progenitors and is defined by the stopping of myeloid lineage development and abnormal proliferation . The mosOxidative stress is implicated in several acute and chronic diseases, including hematological malignancies such as acute myeloid leukemia, which is the most common acute leukemia in adults, with an increasing incidence with age and high relapse rates . Despite2) on days 1–3 and cytarabine (100–200 mg/m2) in continuous infusion on days 1–7 [2/12 h [Cytarabine with anthracycline induction therapy is the standard of care for acute myeloid leukemia. The most commonly used regimen includes anthracycline daunorubicin ; Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS); NRAS Proto-Oncogene, GTPase (NRAS); Tyrosine-protein phosphatase non-receptor type 11 (PTPN11); neurofibromin 1 (NF1); and KIT proto-oncogene, receptor tyrosine kinase (KIT)). It is interesting to note that mutations in this class are usually found in subclonal cellular fractions, indicating that they are frequently late clonal events in the development of illness [Genetic mutations that have familial causes should also be considered . The mos illness .Somatic mutations within key epigenetic regulators are identified in >50% of acute myeloid leukemia, and are now recognized as a key, and often an inciting, component of leukemogenesis . It is oFLT3 and DNA methyltransferase 3 alpha , nucleophosmin 1 (NPM1) is one of the three most frequent driver mutations in acute myeloid leukemia. The regulation of pathways for cell proliferation, differentiation, adhesion, and death by receptor tyrosine kinase (RTKs) signaling pathways in acute myeloid leukemia is crucial for the onset and spread of malignancy. Around 20 separate subfamilies make up the RTKs, which include class III and TYRO3, AXL, and MERTK (TAM) family RTKs [c-Kit, CSF1R, FLT3, and platelet-derived growth factor receptors (PDGFR), have been discovered to have a major effect on leukemogenesis and transformation into acute myeloid leukemia. Class III RTKs have been linked to aberrant activation that promotes proliferation in leukemia. Particularly, FLT3 expression and c-Kit mutations are critical for acute myeloid leukemia. Both RTKs have been crucial targets in the development of antileukemic therapies, since they are both associated with worse prognoses. TYRO3, AXL, and MERTK are members of the TAM family of RTKs. These are essential for platelet activation and stabilization, for the normal hematopoiesis of certain innate immune cells, and have been linked to erythropoiesis [AXL and MERTK, have been linked to hematologic malignancies and have grown in interest as potential targets for creating novel treatments [Along with ily RTKs . Class Iopoiesis . TAM RTKopoiesis . TAM RTKeatments ,56,57,58DNMT3A, TET1/2, and IDH1/2 [In addition, compared to the genomes of other cancers, acute myeloid leukemia genomes are typically less mutated , with sid IDH1/2 . Hassan d IDH1/2  point toIDH1 gene mutations are present in around 6–10% of individuals with acute myeloid leukemia [IDH1 is implicated in controlling cellular metabolism, particularly lipid metabolism [IDH1 Arg132 mutation alters the way the enzyme functions, causing α-ketoglutarate to be converted to R(—)-2-hydroxyglutarate [leukemia . Given itabolism ,66. Isoctabolism . It has lutarate . This exlutarate .NPM1 gene encodes for the nuclear chaperone protein NPM, which shuttles between the nucleus and cytoplasm and plays a role in diverse cellular functions, including protein formation, ribosome biogenesis, DNA replication, and the cell cycle. NPM1 mutations are typically stable throughout the disease course, are identified in nearly all leukemic cells, and impart a distinct expression profile [NPM1 mutations in the setting of mutant DNMT3A, particularly in the setting of FLT3 internal tandem duplication (FLT3-ITD), confer a markedly poor prognosis [Nucleophosmin 1 mutations, occurring almost exclusively within exon 12 of the gene, occur in approximately one-third of adults with acute myeloid leukemia, and in more than 50% of NK-AML. The  profile . NPM1 murognosis ,71.RUNX1), CEBPA, and/or GATA binding protein 2 (GATA2) [RUNX1 is known to act as a direct transcriptional activator of several proteins important for platelet function and as a transcriptional repressor of others, including MYH10 [ANKRD26 [CEBPA encodes a master hematopoietic transcription factor that acts as a critical regulator of granulocyte and monocyte differentiation [GATA2 encodes a zinc finger transcription factor critical for normal hematopoiesis [Approximately 20% to 25% of adults with acute myeloid leukemia have mutations involving the myeloid transcription factors Runt-related transcription factor 1 ( (GATA2) . In thisng MYH10 ,73,74,75[ANKRD26 . In addintiation , while Gopoiesis ,79 and lopoiesis ,81.TP53) is a key tumor suppressor with highly variable functions related to the maintenance of genomic stability, including regulation of cellular senescence, apoptosis, metabolism, and DNA repair. Although uncommon in de novo acute myeloid leukemia, TP53 mutations occur in ~15% of therapy-related acute myeloid leukemia or acute myeloid leukemia with myelodysplastic syndrome-related changes, and are predominantly associated with complex cytogenetics, advanced age, chemotherapy resistance, and poor survival [TP53 mutations in acute myeloid leukemia portend lower response rates and inferior outcomes compared with TP53 wild-type acute myeloid leukemia patients [Tumor protein p53  are identified in ~10% of patients with acute myeloid leukemia and are associated with older age, less proliferative disease, poor rates of response to standard treatment, and decreased survival. Spliceosome mutations are postulated to promote malignancy through the missplicing of various genes involved in epigenetic regulation, transcription, and genome integrity [Frequently mutated in myelodysplastic syndrome and myeloproliferative neoplasms, mutations in splicing factors (ntegrity .SMC3 and SMC1A), RAD cohesin complex component (RAD21), and cohesin subunit SA (STAG1/STAG2) make up the four core elements of cohesin with a ring shape. Cohesin helps several other subunits, such as NIPBL, MAU2, WAPL, PDS5A, PDS5B, and sororin, to form cohesion during the cell cycle [RNAPII), or transcription factors (TFs) [The structural maintenance of chromosomes  ,93,94,95MECOM  complex locus) gene on chromosome 3q26.2 have a very poor prognosis [Acute myeloid leukemia prognosis is exceedingly variable and unpredictable. It can be caused by molecular changes, chromosomal translocations, or genetic mutations. Genetic mutations have been shown to occur in around 97% of cases. rognosis ,105.FLT3, NPM1, CCAAT/enhancer binding protein α (CEBPA), RUNX1, additional sexcombs-like 1 (ASXL1), and TP53 [DNMT3A and TET2 genes, has been linked to leukemia genesis [DNMT3A and TET2 genes are common in patients with clonal hematopoiesis of undetermined potential [Modern molecular technologies have permitted the identification of a wide spectrum of genetic disorders. Six genes have already been incorporated into the European Leukemia Net risk categories , includiand TP53 ,110,111.and TP53 ,117,118. genesis ,120. Mutotential ,121,122;otential ,120. A sCurrently, niches are thought of as microenvironments that mix non-hematopoietic cells with the structure of the bone marrow to encourage hematopoietic stem cell self-renewal and differentiation by offering beneficial and crucial components ,124. TheDue to their ability to directly govern the development and differentiation of hematopoietic stem cells and their ability to release a range of soluble growth factors and cytokines, mesenchymal stromal cells really play an important role in immunomodulation ,126,127.The ability of allogeneic stem cells to differentiate into various stromal marrow components, such as pericytes, bone marrow stromal cells, myofibroblasts, osteoblasts, osteocytes, and endothelial cells, is also essential for allogeneic stem cell transplantation to be successful ,133. BonAdditionally, animal models of myeloproliferative neoplasms ,146,147,PTPN11 mutant was expressed [By using array comparative hybridization and transcriptome profiling, it was also discovered that CD271+ mesenchymal stromal cells directly obtained from myelodysplastic syndrome patients had genetic and transcriptomic changes ,159,160.xpressed .CYP2E1), glutathione S-transferase Mu 1 (GSTM1), NAD(P)H: quinone oxidoreductase (NQO1), N-acetyltransferase 2 (NAT2), and multidrug resistance protein 1 (MDR1), influence leukemia risk alone or in combination with chemical exposure [Despite the above-mentioned, few studies have used environmental sampling ,162 and exposure .CYP2E1 is not only markedly elevated in the liver, but also expressed in bone marrow [Salvia miltiorrhiza, Schisandra chinensis), pollutants (phenylamine), food additives (coffee and cocoa polyphenols), and industrial material and environmental contaminants (benzene) can stimulate CYP2E1 activity [CYPs are a family of heme-containing proteins that play an essential function in the metabolism of a wide variety of xenobiotics . CYP450 e marrow . Drugs aactivity ,171.CYP2E1, is a precondition for the cyto- and genotoxic effects of benzene exposure [Benzene has been recognized as an environmental contaminant that can generate hematotoxicity and leukemogenicity ,173,174.exposure ,176,177.exposure . These pexposure ,180,181.exposure ,183,184 exposure .CYP2E1-induced hepatic oxidative stress, which could be a viable treatment for ethanol-induced fatty liver [CYP2E1 induction and the involvement of PI3K/Akt in guarding against the cytotoxicity of CYP2E1 suggest that CYP2E1 overexpression may reduce the expression of critical proteins in the PI3K signaling pathway [Among the biochemical mechanisms abnormally elevated in malignancies, including acute myeloid leukemia, is the phosphoinositide 3-kinase-Akt-mammalian target of rapamycin route (PI3K-Akt-mTOR pathway) . PI3K enty liver ,191. The pathway ,191.CYP450 enzymes serve a critical role in preventing oxidative damage; they metabolize several exogenous and endogenous genotoxic chemicals, such as hydrogen peroxide, by inserting an oxygen atom into the substrate .CYP450 loci  may very well be categorized as risk factors for numerous kinds of cancers, due to the inactivation of enzymatic activity [Specific single-nucleotide polymorphisms at the activity ,199,200.CYP2B6 gene locus, the G516T polymorphism has been recognized as a nonsense polymorphism that decreases the activity of the protein complex. Hence, people who have the T allele (TT) have a reduced enzymatic activity than people who have the wild-type G allele (GG), but individuals holding the genotype GT exhibit intermediate activity [At the activity .CYP3A enzyme activity to vary up to 50-fold. Most genetic variants in the CYP3A4 gene result in lower enzyme activity. Single-nucleotide polymorphisms in the CYP3A4 promoter region (A290G)  have been implicated as a possible cause of this variability; hence, it may be considered a risk factor for cancer. However, the implications of these single-nucleotide polymorphisms are not well-understood [CYP450 gene have distinct pathogenic effects and prognostic characteristics in various hematological cancers. Ethnicity causes derstood ,204,205.The broad category of xenobiotics includes substances that are generally safe but may be harmful, such as medications, environmental toxins, cosmetics, and even elements included in our diet, such as food additives ,208,209.Xenobiotic metabolism increases their water solubility, thus enhancing their elimination from the body . When xeThe inflow of xenobiotics is mediated by sodium taurocholate cotransporting polypeptide, organic anion transporting polypeptides, and organic anion transporters . Phase IThere are significant inter- and intra-individual differences in the ability to metabolize, detoxify, and expel xenobiotics. These are genetic, epigenetic, environmental, and physiological pathophysiological in nature, and they change during life ,229,230.CYP2E1 is a hepatic monooxygenase involved in the metabolism of xenobiotics. The CYP2E1 gene is linked to the metabolism of many carcinogens. CYP2E1 is essential for the metabolism of endogenous compounds (such as acetone and fatty acids) as well as external substrates such as medications, contaminants, and ethanol [CYP2E1-mediated metabolism  and enzyme-associated toxicity, such as methemoglobinemia and acetaminophen-induced liver necrosis, has recently piqued researchers’ attention. Electrons are transferred to the substrates through CYP2E1-mediated oxidation of reduced nicotinamide, adenine dinucleotide phosphate, and molecular oxygen. This reaction adds additional polar groups to the substrates and generates hazardous intermediates such as epoxides or aldehydes [ ethanol . The mecldehydes .CYP2E1 active site as a sheet of hexagonal blocks has been devised, which might directly relate to the two-dimensional structure of chemicals. For a core part of the CYP2E1 active site, a region with the shape of benzopyrene was suggested as a model [An in silico approach for simulating the  a model . This waCYP2E1 polymorphisms have been linked to potential mechanisms of tumor initiation. The CYP2E1*5 allele is linked to an increased risk of developing acute myeloid leukemia and acute lymphoblastic leukemia [CYP2E1 single-nucleotide polymorphisms rs2070673TT and rs2030920CC are exposed to phenol, they exhibit increased transcription and enzyme activity as well as increased DNA damage [CYP2E1*5B(C-1019T) polymorphism has not been linked to therapy-related acute myeloid leukemia or myelodysplastic syndrome [CYP2E1*07 (rs2070673) allele has been linked to a higher survival rate [CYP2E1 gene have diverse pathogenic and prognostic consequences in various hematological malignancies. Furthermore, the existence of polymorphisms does not always correspond with the phenotypic functional activity of CYP2E1, and total functional evaluation is more accurate than testing for polymorphisms. Increased CYP2E1 expression has been linked to liver illnesses such as alcoholic hepatitis and non-alcoholic steatohepatitis and is considered to play a role in their etiology [leukemia . When lyA damage . The CYPsyndrome . In indival rate . As a reetiology ,240.MLL) gene rearrangements [NQO1 activity. These findings support the idea that common ambient pollutants detoxified by NQO1 are risk factors for acute leukemia [NQO1 polymorphism had the strongest connection with acute myeloid leukemia and inv(16)/CBF-MYH11 in a study concentrating on de novo acute myeloid leukemia [NQO1 gene, which is located on chromosome 16q22.1 [CYP2E1 was one of the four most differentially expressed genes in acute myeloid leukemia with inv(16)/CBF-MYH11, being raised 3.3-fold in acute myeloid leukemia with inv(16) [Therapy-related myeloid neoplasms , infant ngements , and a sngements , have loleukemia . In factleukemia . While t 16q22.1 , it is a inv(16) .Between the treatments used for acute myeloid leukemia, we can find azacitidine, which is a pyrimidine nucleoside analog of cytidine that can be directly integrated into RNA, disrupting RNA, protein production, and metabolism . It is oAzacitidine is available for intravenous and subcutaneous injection, with equivalent absorption for both routes . The cytCYP3A4 [Other treatments for acute myeloid leukemia are based in enasidenib, a novel, mutant IDH2 protein-targeting inhibitor used to treat relapsed or resistant acute myeloid leukemia ,252,253.CYP3A4 . Since eCYP3A4 . HoweverCYP3A4 .CYP3A4, with a minor contribution from glucuronidation by uridine diphosphate glucuronosyltransferase 1A [CYP3A4 enzyme-mediated metabolism, plasma levels of glasdegib tend to decline; CYP3A inhibitors such as ketoconazole should be administered to increase glasdegib levels [Finally, drugs against acute myeloid leukemia, such as glasdegib, gilteritinib, midostaurin, cytarabine, and venetoclax, have high oral bioavailability and are widely used ,259,260.erase 1A . Howeverb levels . Table 3Dietary phytochemicals are one specific class of dietary components having anti-cancer action, among other dietary variables that are well known for their chemo-preventive effects. In addition to being non-essential nutrients, phytochemicals can significantly contribute to the prevention of disease. Numerous phytochemicals have strong anti-oxidant and anti-carcinogenic properties, including polyphenols, flavonoids, allyl sulphides, and carotenoids .CYP2E1 activity after moderate alcohol consumption and β-carotene supplementation [Carotenoids are pigments found in fruits, vegetables, and whole grains that are yellow, orange, or red. Patients with asthma, cataracts, and heart disease have been shown to benefit from beta-carotene, the main source of vitamin A and its derivative retinoic acid ,273,274.entation , other sentation ,277. TheCYP2E1 in the xenobiotic metabolism that governs the myeloid leukemia microenvironment. Still, this review found some problems that could make it hard to combine scientific evidence. These problems include: (1) a lack of information because there were so few articles; (2) a lack of NQO1 levels during CYP2E1 activity in acute myeloid leukemia; (3) a lack of information about how gene polymorphisms affect the encoded protein; and (4) a lack of thought about how genes and the environment interact.The purpose of this review was to describe the pathophysiology of acute myeloid leukemia as well as the role of CYP2E1 contributes to an elevated acute myeloid leukemia risk. CYP2E1-related disorders relate to protein levels, and there are inter-individual variances in CYP2E1 expression levels, according to research. Furthermore, genetic polymorphism, drugs, plants, pollutants, food additives, and industrial material and environmental contaminants influence the variability and susceptibility to related myeloid neoplasms, infant leukemias associated with MLL gene rearrangements, and a subset of de novo acute myeloid leukemia. Recent research has shown a sustained interest in determining the regulators of CYP2E1 expression and activity as an emerging field that requires further investigation in acute myeloid leukemia evolution. This research has the potential to give insight into novel strategies for the treatment of acute myeloid leukemia via CYP2E1.Through the conversion of a range of xenobiotics into hazardous intermediates such as reactive oxygen species and free radicals,


This study examined the relationship between levels of the chemokines CXCL9, CXCL10, CXCL11, and CXCR3 and mortality in patients with COVID-19.. A total of 71 patients hospitalized with COVID-19 and 35 health workers with no symptoms and negative SARS-CoV-2 PCR results were included in the study. CXCL9, CXCL10, CXCL11, and CXCR3 levels were measured in blood samples using enzyme-linked immunosorbent assays. Participants were divided into three groups: healthy individuals, patients with mild to moderate pneumonia, and patients with severe pneumonia. Patients were also divided into sub-groups according to the outcome: dead and survived.2/FiO2 levels were significantly lower in patients with severe COVID-19 than in those with moderate disease. Patients with COVID-19 also had elevated neutrophil/lymphocyte ratios, neutrophil counts, and lactate dehydrogenase, C-reactive protein, D-dimer, and ferritin levels. Serum CXCL9, CXCL10, CXCL11, and CXCR3 levels were significantly higher in patients with severe COVID-19 than in those with non-severe COVID-19; were higher in both patient groups than in the control group; and were higher in patients who died than in those who survived. Lymphocyte counts, and fibrinogen and PaO This study confirmed that CXCL9, CXCL10, CXCL11, and CXCR3 levels are associated with disease severity in patients with COVID-19. These laboratory parameters can help to estimate disease severity and predict outcomes, and are useful in clinical decision-making. COVID-19 is a potentially fatal viral disease, ranging in severity from mild upper airway infection to severe pneumonia, acute respiratory distress syndrome (ARDS), and multiple organ failureA hyperinflammatory host response plays an important role in the manifestation of severe diseaseCytokines are a category of proteins, peptides, and glycoproteins secreted by hematopoietic and non-hematopoietic cells that respond to different types of stimuli. They are categorized into five groups: the interleukin-1, hematopoietin, interferon, tumor necrosis factor, and chemokine familiesChemokines are important proteins involved in the regulation of leukocyte migration in peripheral lymphatic organs and play an important role in the inflammatory response. They cannot be detected in most nonlymphoid tissues under physiological conditions. Their production by blood and tissue cells is strongly induced by interferon gamma (IFN-γ), the most typical T helper 1 (Th1) cytokine in several diseasesGenerally, chemokines contribute to combating viral infections by recruiting immune cells to the locus of infection, increasing the cytotoxic activities of these cells, and encouraging the release of antiviral mediatorsCXCL9, CXCL10, and CXCL11 are three chemokines without the Glu-Leu-Arg (ELR) motif. Due to their approximately 40% amino acid sequence similarity, they are more closely related than other chemokines. These chemokines play a specific role in the Th1 responseThe primary aim of this study was to investigate the association between mortality and the CXCL9, CXCL10, CXCL11, and CXCR3, chemokine levels, which contribute to the pathogenesis of COVID-19 through hyperinflammation, in patients with COVID-19.Seventy-one patients with COVID-19 hospitalized in the COVID-19 ward of our hospital between January and September 2021 and 35 healthcare workers with negative reverse-transcription polymerase chain reaction (RT-PCR) SARS-CoV-2 test results were included in this study. Patients aged older than 18 years were enrolled; however, those with other acute or chronic systemic disease, hematologic disease, malignancy, or immunosuppressive drug use were excluded. The diagnosis of COVID-19 was confirmed by a positive SARS-CoV-2 RT-PCR results on a nasal swab specimen, or chest radiography and computed tomography findings compatible with COVID-19. This study was conducted in accordance with the principles of the Declaration of Helsinki and was approved by the Health Sciences University Erzurum Regional Education and Research Hospital Clinical Research Ethical Committee, Türkiye (2022/08-108).2 ≤ 92%, and/or pulmonary infiltration > 50%) (n = 41). The patients were also subdivided into those who died and those who survived. Two groups were also established based on the extent of lung involvement on imaging: mild/moderate involvement and severe involvement. The extent of pulmonary involvement was classified as minimal (1-5%), mild (6-25%), moderate (26-49%), severe (50-74%), and diffuse (≥ 75%). Serum levels of CXCL9, CXCL10, CXCL11, and CXCR3 were compared between groups.The 106 individuals enrolled in this study were assigned to three groups. The first group consisted of healthy PCR-negative healthcare workers who consented to participate (n = 35). The second group consisted of patients with moderately severe pneumonia (n = 30). The third group consisted of patients with severe pneumonia  and ferritin levels, D-dimer elevation, lymphopenia or thrombocytopenia, abnormal liver function tests, or hypofibrinogenemia were monitored for macrophage activation syndrome (MAS).Blood specimens were centrifuged for 15 min at 4000 rpm to separate the serum. The serum samples were stored in Eppendorf tubes at −80ºC. CXCL9, CXCL10, CXCL11, and CXCR3 levels were measured in the same blood specimen and recorded. Laboratory test results, including serum CRP, lactate dehydrogenase (LDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyltransferase (GGT), creatinine, D-dimer, troponin I, prothrombin time (PT), fibrinogen, ferritin, white blood cell (WBC), lymphocyte count, neutrophil count, neutrophil/lymphocyte ratio, and platelet count were also recorded.Serum CXCL9, CXCL10, CXCL11, and CXCR3 levels were measured using an enzyme-linked immunosorbent assay . Serum LDH, AST, ALT, ALP, GGT, CK, creatinine, and CRP levels were measured using commercial kits and a Beckman Coulter AU5821 analyzer . Ferritin and troponin I levels were measured using commercial kits and a Beckman Coulter DxI800 analyzer. WBC, lymphocyte, neutrophil, and platelet counts were measured using a Sysmex XN-9000  analyzer. D-dimer levels were measured using a Radiometer AQT90 flex analyzer . The activated partial thromboplastic time, international normalized ratio, and fibrinogen level were measured using a Stago STA R Max analyzer .Data were analyzed using SPSS for Windows . Categorical variables were expressed as frequencies and percentages, and continuous variables were expressed as means ± standard deviations. Visual (histogram) and analytical methods  were used to assess the normality of the distribution of continuous variables. Chi-square tests were used to compare categorical variables between groups, and t-tests or one-way analysis of variance were used to compare continuous variables between groups, as required. The significance of intergroup differences was evaluated using post-hoc Tukey tests. Receiver operating characteristic (ROC) curves were plotted, and the optimal cut-off values sensitivity, specificity, and area under the curve (AUC) were calculated to assess the predictive power of different measures. Statistical significance was set at 5%.Seventy-one patients with COVID-19 and 35 healthy controls were included in the study. The mean age was 65.8 ± 14.5 years in the patient group and 62.6 ± 12.3 years in the control group. No significant differences in age (p = 0.41) or sex (p = 0.23) were observed between the groups.The COVID-19 group consisted of 45 men (63%) and 26 women (37%). Among the men, 21 (47%) had moderate COVID-19 and 24 (53%) had severe COVID-19. Among the women, 9 (35%) had moderate COVID-19 and 17 (65%) had severe COVID-19. COVID-19 severity did not differ significantly by sex (p = 0.07). The mean age was 65.53 ± 15.89 years in patients with moderate disease and 66.05 ± 13.12 years in those with severe disease. COVID-19 severity did not differ significantly by age (p = 0.88).In the COVID-19 group, the survivors were aged 65.74 ± 12.92 years, and those who died were aged 65.92 ± 16.22 years. Men comprised 27 (64%) of the survivors and 19 (66%) of the patients who died. No differences were observed in COVID-19 severity by age or sex .2/FiO2 levels were all significantly lower in patients with severe COVID-19 than in those with moderate disease . The neutrophil/lymphocyte ratio (NLR), neutrophil count, and LDH, CRP, D-dimer, and ferritin levels were all significantly higher in patients with severe COVID-19 than in those with moderate disease .The laboratory data of patients with COVID-19 are compared by disease severity in . CXCL9, CXCL10, CXCL11, and CXCR3 levels were significantly higher in patients with severe COVID-19 group than in those with moderate disease. CXCL9, CXCL10, CXCL11, and CXCR3 levels were significantly higher in both patient groups than in the control group.The CXCL9, CXCL10, CXCL11, and CXCR3 levels in the COVID-19 group and healthy controls are compared in Table 2The CXCL9, CXCL10, CXCL11, and CXCR3 levels were significantly associated with clinical manifestations including fever, tachypnea, muscle/joint pain, sore throat, headache, somnolence, shortness of breath, and nausea/vomiting .The patients with COVID-19 were also divided into two groups according to the severity of pulmonary involvement on chest imaging. The first group consisted of patients with mild involvement, and the second group consisted of patients with severe involvement. CXCL9, CXCL10, CXCL11, and CXCR3 levels were compared between the two groups . CXCL9, The patients were further classified into two groups based on the outcome: died or survived. Patients who died were more likely to develop MAS during follow-up. CXCL9, CXCL10, CXCL11, and CXCR3 levels were significantly higher in patients who died .In the ROC curve analysis of the ability of CXCL9, CXCL10, CXCL11, and CXCR3 to discriminate between moderate and severe COVID-19, the AUC values were 0.702, 0.939, 0.933, and 0.857, respectively. Using cut-off values of 689.1, 228.4, 248.1, and 688.1 pg/mL according to the Youden index elicited sensitivity values of 63%, 83%, 81%, and 78% and specificity values of 70%, 90%, 90%, and 83%, respectively .,In addition to their function in the regulation of leukocyte migration, chemokines and their receptors are involved in a broad range of pathophysiological processes such as inflammation, infectious diseases, allergic reactions, autoimmune diseases, and cancerCOVID-19 is a highly heterogeneous viral disease, and is characterized by a clinical course ranging from asymptomatic to fatal. Once the pathological role of the cytokine storm during COVID-19 became recognized, studies investigated whether specific chemokines may play a role in its progression,CXCL10 can be released by different cells, including monocytes, leukocytes, and endothelial and epithelial cells. CXCL10 is elevated in several diseases, including hepatitis B, tuberculosis, cancer, diabetes, and autoimmune disorders,+ T-cell countIn agreement with previous studies, this study found higher CXCL10 levels in patients with severe pneumonia than in patients with moderate pneumonia, and significant elevation in patients developing MAS and fatal cases. CXCL10 levels have also been shown to be increased in patients with ARDS. However, higher CXCL10 levels were found in patients with ARDS manifestations thought to be of viral origin than in those with ARDS of suspected bacterial origin,CXCL9 is induced by IFN-γ in macrophages and is involved in cancer and viral infections. It is also thought to play a role in T-cell trafficking, chemotaxis, and activation+ T-cell count in these diseases,,,CXCL11 is expressed at high levels in peripheral blood leukocytes, the pancreas, and the liver. Of CXCL9, CXCL10, and CXCL11, CXCL11 exhibits the greatest affinity for CXCR3, followed by CXCL10 and CXCL9+ Th1 cells and CD8+ T cells to the site of tissue injury, and inhibit angiogenesis during pulmonary fibrosis. All three chemokines have been implicated in the pathogenesis of idiopathic pulmonary fibrosis. Most patients with severe COVID-19 develop pneumonia and hyperinflammation, probably as a result of the MAS, known as a cytokine storm. Pulmonary fibrosis occurs as a secondary event linked to the progression of pathology associated with the inflammatory responseCXCR3 is a receptor that is preferentially expressed on the surfaces of monocytes, T cells, NK cells, dendritic cells, and cancer cellsIn this study, the levels of these chemokines were compared according to the severity of pulmonary involvement. Significant elevations in the levels of all three chemokines and their receptors were observed in the severe pulmonary involvement group. This finding suggests that these factors may be responsible for the development of pulmonary fibrosis in patients with severe COVID-19.,,Increases occur in parameters such as AST, ALT, urea, creatinine, LDH, creatine kinase, troponin, and CK-MB occur secondary to organ damage as the disease progresses and organ dysfunction develops. As in several other infections, an increase occurs in the CRP and ferritin levels, and the erythrocyte sedimentation rate as the disease progresses, and this has been linked to severe lymphopenia, thrombocytopenia, and leukopenia2/FiO2 levels were significantly lower in patients with severe COVID-19, whereas WBC, NLR, LDH, CRP, D-dimer, PT, and ferritin values were significantly higher in those with severe disease.The results of this study are consistent with the results of previous studies in terms of most parameters. Lymphocyte counts, fibrinogen levels, and PaOIn this study, CXCL9, CXCL10, CXCL11, and CXCR3 levels showed higher specificity than sensitivity based on the Youden index. This suggests that these chemokines may be effective in predicting the course of severe COVID-19.Interferon-inducible chemokines such as CXCL9, CXCL10, and CXCL11 may be vital to the host’s response in viral respiratory infections by encouraging viral elimination before the activation of the adaptive immune systemThe principal limitation of this study is the absence of a control group of patients with other illnesses associated with pneumonia, such as influenza or bacterial pneumonia. Such a control group would clarify whether the observed differences in the humoral immune response are specific to SARS-CoV-2. Further studies involving control groups are required to determine the usefulness of these chemokines in identifying COVID-19.In conclusion, identifying potential targets for reducing chemokine induction could be an important strategy for treating viral respiratory infections. This study emphasizes the association between CXCL9, CXCL10, CXCL11, and CXCR3 levels and disease severity in patients with COVID-19. It shows that these can predict disease severity and outcome and could assist in clinical decision-making. These findings may serve as a guide for future research. This study contributes to the literature by confirming the relationship between chemokine levels and fatal COVID-19 in a more accurate manner, and by supporting previous studies on the subject.


This scoping review aimed to summarize the effects of natural products targeting phosphoinositide-3-kinases/serine/threonine kinase (PI3K/AKT) in myocardial ischemia-reperfusion injury (MIRI). The review details various types of natural compounds such as gypenoside (GP), gypenoside XVII (GP-17), geniposide, berberine, dihydroquercetin (DHQ), and tilianin which identified to reduce MIRI in vitro and in vivo by regulating the PI3K/AKT signaling pathway. In this study, 14 research publications that met the inclusion criteria and exclusion criteria were shortlisted. Following the intervention, we discovered that natural products effectively improved cardiac functions through regulation of antioxidant status, down-regulation of Bax, and up-regulation of Bcl-2 and caspases cleavage. Furthermore, although comparing outcomes can be challenging due to the heterogeneity in the study model, the results we assembled here were consistent, giving us confidence in the intervention’s efficacy. We also discussed if MIRI is associated with multiple pathological condition such as oxidative stress, ERS, mitochondrial injury, inflammation, and apoptosis. This brief review provides evidence to support the huge potential of natural products used in the treatment of MIRI due to their various biological activities and drug-like properties. Coronary heart disease is the third leading cause of global mortality which associated with 17.8 million of deaths yearly . TherefoNatural products (NP) derived from plants could be an alternative source of promising drugs to mitigate the harmful aftereffects of MIRI. NP offers various medicinal values and health benefits ,10,11,12The phosphoinositide-3-kinases/serine/threonine kinase (PI3K/AKT) is an intracellular signaling pathway involved in the regulation of cellular metabolism, survival, and apoptosis . Many stAccordingly, the damages related to MIRI are reduced when the PI3K/AKT pathway is regulated, implying its significance in cardioprotection ,14,15. An = 36), combination of herbs in the treatment (n = 2), and no availability of full text (n = 1). Thus, a total of 14 articles from January 2016 to January 2021 were selected in this scoping review. A flowchart depicting the search process and study selection is presented in Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) for Scoping Review , other signaling pathways such as PI3-kinase/glycogen synthase kinase 3β/β-catenin pathway, NFkB signaling, IGF-1R/Nrf2 signaling, and the PI3k/Akt signaling pathway were not measured. By full-text screening of the remaining article 53 articles, a total of 39 articles were eliminated based on the inclusion/exclusion criteria; study models used were not MIRI experiment models  of the animals were ligated to induce ischemia for some period. Subsequently, ligation was released to mimic reperfusion. Even though the research adopted a similar in vivo design, the timings for ischemia and reperfusion were different. For example, Yu et al., 2016, ligated and occluded LAD for 45 min followed by 3 h of reperfusion, Jiang et al., 2016, occluded the LAD for 30 min followed by 4 h of reperfusion, Zeng et al., 2018, introduced ischemia for 45 min, followed by 4 h of reperfusion, Zeng et al., 2018, and Zhao et al., 2018, ligated the LAD for 45 min mimicking ischemia, followed by 12 h of reperfusion and Wu et al., 2016, induced ischemia by ligation of LAD for 30 min, followed by 24 h of reperfusion.Additionally, IR can also be induced with prolonged cold ischemia (18 h) and followed by 8 h of reperfusion in rat. For the in vitro model, hypoxic-reoxygenation (H/R) injury was established by culturing H9c2 cell in glucose-free DMEM medium placed in the anaerobic glove box or hypoxic chamber containing high percentage of nitrogen gas to mimic hypoxia. Then, the medium of the cells was changed with glucose containing medium and placed in the regular incubator with atmosphere of 95% air. Using similar H/R injury method in H9c2 cells, Wu et al., 2013, induced 8 h of hypoxia followed by 4 h of reoxygenation, Shen et al., 2018, induced 10 h of ischemia followed by 2 h of reperfusion, and Liao et al., 2016, induced 6 h of hypoxia and 12 h of reoxygenation.In addition, MIRI can be induced ex vivo using Langendorff perfusion model. There were 3 studies using this model. A study by Mei-ping Wu et al., 2016, stopped the perfusion on the isolated heart for 30 min to induce ischemia and then reperfusion for another 30 min, Zun Peng Shu et al., 2019, found the perfusion on the isolated heart was stopped for 45 min to induce cardiac ischemia and then reperfusion occurred for another 60 min, while Yingli Yu et al., 2019, introduced global ischemia for 40 min and reperfusion for 60 min.Dracocephalum moldavica was extracted in ethanol [Dracocephalum moldavica was further extracted in aqueous ethanol and purified to obtain tilianin, which was then identified by HPLC [Five studies mentioned the part of the plant, method of extraction, and the solvents used ,23,26,28 ethanol ,23. HPLC by HPLC . Whereas by HPLC ,30,31,32Gynostemma pentaphyllum (Thunb.) Makino [Gypenoside is the main component of ) Makino ,34, and ) Makino . In an o) Makino . In the ) Makino .Gypenoside XVII (GP17) is an antioxidant phytoestrogen of the gypenosides group has a similar structure as estradiol ,36. GP17Gardenia jasminoides J. Ellis inhibited cardiomyocytes apoptosis, thus improving cell viability and decreased LDH levels [In a hypoxia/reoxygenation (H/R) of H9c2 cells, pre-treatment with geniposide, an extract from H levels . MitochoH levels .Berberis vulgaris L. [Berberine is an alkaloid derivative, isolated from Coptis chinensis Franch  and Berbgaris L. . In the garis L. . In a segaris L. .Allium cepa L. and Silybum marianum (L.) Gaertn [Dihydroquercetin (DHQ) is a flavonoid that is commonly found in ) Gaertn . In H9c2) Gaertn . Two myo) Gaertn .Dictamnus dasycarpus Turcz [Cortex Dictamni (CD) is a species of us Turcz ,41. Aqueus Turcz .Dracocephalum moldavica L. is a herbaceous plant that belongs to the Lamiaceae family [Dracocephalum moldavica (TFDM) was found to decrease MIRI severity by decreasing CK-MB, LDH, MDA, and apoptotic cells index at aa dose of 3, 10, and 30 mg/kg. Furthermore, it was also able to reduce the infarction size, preserved cell membrane, and mitochondria. In addition, antioxidative and anti-apoptotic properties of TFDM were confirmed by increased SOD and Bcl-2/Bax ratio and decreased cleaved caspase-3, -7 and -9 levels. Additionally, TFDM elevated p-PI3K and p-AKT as well p-PI3K/PI3K and p-AKT/AKT ratio. Activation of PI3K/AKT signaling was facilitated by protein expression of p-GSK-3β and p-ERK1/2. The cardioprotective effect of TFDM was abolished upon administration of LY294002 and PD98059, which were the inhibitor of PI3K and ERK 1/2 phosphorylation, respectively [e family . Total fectively .Dracocephalum moldavica L.(DML) [Tilianin is a powerful flavonoid in dried  L.(DML) . Tiliani L.(DML) .Schisandra chinensis (Turcz.) Baill. [Schisandrin B (Sch B) is a natural medicinal monomer extracted from ) Baill.  Sch B re) Baill. . AKT act) Baill. ,45.2O2-induced cardiomyocyte apoptosis. The protective effect of higenamine was achieved via upregulation of cleaved-caspase-3 and -9 and protein expression of p-AKT. However, ß2-adrenergic receptor (AR) antagonist and PI3K inhibitor eliminated these protective effects [Higenamine derives from plant-based protoberberines alkaloids . Higenam effects . In summMyrica rubra (Lour) Siebold and Zucc. belongs to the family Myricaceae. Hypoxic/reoxygenation of H9c2 cells revealed that Myrica rubra flavonoid (MRF), at a dose of 6.25 µg/mL, was able to attenuate intracellular ROS level, protected the cells against mitochondrial membrane potential depolarization, inhibited caspase-3 activity, and alleviated the ratio of apoptotic cells [ic cells . This pric cells .Paeonia lactiflora Pall. [Total paeony glycoside (TPG) is a major constituent of ra Pall. . TPG redra Pall. .Bauhinia championii Benth. or Phanera championii Benth. belongs to the genus Bauhinia [Bauhinia championii flavone (BCF), at a dose of 3.125 µg/mL, markedly attenuated apoptotic rate, caspase-3, Bax, and cytochrome-C. BCF also significantly increased the protein expression of p-PI3K and p-AKT. In addition, BCF significantly decreased the level of ROS-generating mitochondria, attenuated mitochondrial transmembrane potential (∆Ψm) dissipation, reduced the mPTP opening and increased ATP production. However, co-administration of LY294002 eliminated the protective effect of BCF [Bauhinia . Bauhinit of BCF . Figure The studies reported in this review utilized Western blotting analysis for detection of PI3K and Akt. Western blot is not only able to detect the protein expression but also able to quantitate protein phosphorylation, which is the fundamental mechanism to study cell signaling . AnalysiActivation of PI3K/Akt signaling involved in the protection against MIRI-induced apoptosis. In order to evaluate the cardioprotective effect played by PI3K/Akt signaling on cell survival, its effects in the apoptotic-related mechanism were observed. There were many assays with various techniques used to measure various aspect of apoptosis. Some studies used flow cytometric quantitation of cell death using Annexin-V/propidium staining to detect early and late apoptotic events. Annexin has greater specificity to bind to the phosphotidylserine (PS), which is the membrane phospholipid. As apoptosis failed to maintain the phospholipid in the cell membrane, PS appear on the outer leaflet of the cell membrane which will be stained with Annexin V. Cells stained positive with Annexin V and negative for PI was suggestive for early apoptotic event, whereas Annexin V positive and PI negative population indicate late apoptotic event . AnotherThe antagonizing effect of PI3K/Akt inhibitors against NP were important to be verified to confirm the PI3K/Akt signaling pathway involvement. Protective effects of NPs in this review were abolished upon administration of PI3K inhibitors such as LY 294002, LY 204002, and wortmannin. There were two studies that did not use PI3K inhibitor ,27 Tabl. PI3K inThis scoping review suggests the evidence of the NP-based therapies that provide cardioprotection in MIRI targeting the PI3K/AKT mechanism. This became a novel therapeutic strategy in the treatment of MIRI. There are increasing numbers of NP-derived drugs used in medicine due to their various biological activities and drug-like properties. One of the major metabolites in the plant is a flavonoid. Previous studies showed that a high intake of dietary flavonoids and their subclasses were associated with lower mortality of cardiovascular disease . The NPsMIRI is associated with multiple pathological condition such as oxidative stress, ERS, mitochondrial injury, inflammatory process, and apoptosis. Therefore, the role of PI3K/Akt signaling pathway post MIRI is very crucial, as it improves the cell survival and viability by limiting the apoptotic event. Generally, the apoptotic signaling pathway can either be stimulated by intrinsic or extrinsic pathways . Bcl-2 aERS is a type of oxidative stress which induces cell dysfunction in MIRI via regulation of the PI3K/AKT pathway . Oxygen Nrf2 is a redox-sensitive transcription factor that plays vital role to combat oxidative stress . Nrf2 coPI3K/AKT pathway is a key signaling cascade that mitigates MIRI. Most of NPs in A systematic search was conducted to identify the NP that alleviate MIRI targeting on PI3K/AKT pathway using PRISMA framework for scoping review . Peer-reThe findings from this scoping review were restricted by several factors. First, our search strategy was focused solely on targeting one signaling pathway, namely the PI3K/Akt pathway, while MIRI is a complex injury that involves various other pathways such as TGFβ1/Smads signaling pathway , Parkin-We summarized that NPs enriched with various bioactive compounds are beneficial to mitigate the pathological effects in post-MIRI. Recent studies suggest that these harmful effects can be alleviated by targeting the PI3K/AKT pathway, which might reduce apoptosis, inflammation, and oxidative stress. Further research is warranted to validate the safety and efficacy of NPs-based drugs in MIRI.


Small-cell lung cancer (SCLC) represents a highly aggressive form of lung cancer. Although SCLC initially responds to chemoimmunotherapy, it inevitably relapses. Antibody–drug conjugates (ADCs) have emerged as a promising treatment approach. ADCs consist of antibodies targeting specific tumor antigens, delivering cytotoxic drugs directly to cancer cells, and reducing side effects. In this review, we collected the available data regarding ADCs evaluated in SCLC. While some ADCs yielded negative results, others, such as those targeting B7-H3, SEZ6, CEACAM5, and TROP2 ADC, show promise. Further research is needed to determine ADCs’ role in SCLC treatment.Small-cell lung cancer (SCLC) is a highly aggressive disease, accounting for about 15% of all lung cancer cases. Despite initial responses to chemoimmunotherapy, SCLC recurs and becomes resistant to treatment. Recently, antibody–drug conjugates (ADCs) have emerged as a promising therapeutic option for SCLC. ADCs consist of an antibody that specifically targets a tumor antigen linked to a cytotoxic drug. The antibody delivers the drug directly to the cancer cells, minimizing off-target toxicity and improving the therapeutic index. Several ADCs targeting different tumor antigens are currently being evaluated in clinical trials for SCLC. Despite the negative results of rovalpituzumab tesirine (Rova-T), other ADCs targeting different antigens, such as B7-H3, seizure-related homolog 6 (SEZ6), and CEACAM5, have also been investigated in clinical trials, including for SCLC, and their results suggest preliminary activity, either alone or in combination with other therapies. More recently, sacituzumab govitecan, an anti-TROP2 ADC, demonstrated promising activity in lung cancer, including SCLC. Furthermore, an anti-B7-H3 (CD276), ifinatamab deruxtecan (DS7300A), showed a high response rate and durable responses in heavily pretreated SCLC. Overall, ADCs represent an intriguing approach to treating SCLC, particularly in the relapsed or refractory setting. Further studies are needed to determine their efficacy and safety and the best location in the treatment algorithm for SCLC. In this review, we aim to collect and describe the results regarding the past, the present, and the future of ADCs in SCLC. Small-cell lung cancer (SCLC) is a highly aggressive neuroendocrine carcinoma, constituting around 13–15% of lung cancer cases. Patients often face an unfavorable prognosis, with a low 5-year survival rate ) . SimilarAnother phase 3 study  was terminated early due to the lack of survival benefits and a high rate of adverse events (grade ≥ 3) . In the A phase 1 study evaluated the safety, tolerability, pharmacokinetics, and preliminary efficacy of SC-002, an ADC directed against DLL3 and conjugated with PBD, in patients with advanced SCLC and large-cell neuroendocrine carcinoma. Among the 35 enrolled patients who received at least one dose of SC-002, 66% experienced serious adverse events, with 37% considered related to SC-002. Grade 3 and 4 adverse events were observed in 60% and 6% of patients, with effusion and hypoalbuminemia being the most common. One patient experienced a grade 5 adverse event. Partial responses were achieved in 14% of patients, while no complete responses were observed . In lineB7-H3, also known as CD276, is a member of the B7 superfamily of immune checkpoint molecules. B7-H3 is a transmembrane protein with an extracellular domain composed of two identical domains (4IgB7-H3 isoforms) and a short intracellular tail . This muB7-H3 is overexpressed in a broad spectrum of cancer histologies. This overexpression appears to be associated with advanced tumor stages and higher tumor grades in various cancers, such as endometrial, cervical, breast, and kidney cancer and oral squamous cell carcinoma. In this light, this elevated expression of the B7-H3 protein has been linked to increased cancer cell proliferation and enhanced invasive potential in several cancer types, including pancreatic, breast, colorectal, and prostate cancer.Interestingly, the role of B7-H3 in predicting patient outcomes is still not clear. In many cancer types, its overexpression has been consistently associated with a less favorable prognosis ,40,41. HIfinatamab deruxtecan  is a targeted ADC against B7-H3, composed of humanized anti–B7-H3 IgG1 conjugated to deruxtecan (Dxd), a topoisomerase I inhibitor, through a cleavable tetrapeptide linker. Upon binding to B7-H3, the ifinatamab deruxtecan complex undergoes internalization and linker cleavage within the intracellular environment by lysosomal enzymes that are overexpressed in tumor cells. Following its release, DXd, which is permeable to cell membranes, induces DNA damage and cell death through apoptosis . DXd, a At present, two studies are currently investigating I-Dxd in SCLC. A phase 1/2 study (NCT04145622) is being conducted in the United States and Japan and focuses on I-Dxd in patients with specific advanced solid tumors (including SCLC) . The stuAs of 31 January 2023, of the 21 SCLC patients, 11 demonstrated an ORR, accounting for 52% of the cohort, comprising 1 complete response (CR) and 10 partial responses (PRs). The median duration of response (DOR) was 5.9 months (95% CI 2.8–7.5). Median PFS was 5.8 months , while median OS reached 9.9 months  .n = 2; grade 2, n = 4; grade 5, n = 1) were adjudicated as drug-related ILD [At the June 30, 2022, data cutoff, the 16.0 mg/kg cohort was closed for safety reasons, with high rates of serious and grade ≥3 treatment-emergent adverse events (TEAs). Anemia (19%), neutropenia (4%), nausea (3%), pneumonia (3%), and decreased neutrophil count (3%) were the most common (23%) grade ≥3 TEAs. ILD/pneumonitis was present in nine patients, of which seven  is currently evaluating the efficacy, safety, and pharmacokinetics of I-Dxd in patients with pretreated (at least one prior line of platinum-based chemotherapy and a maximum of three prior lines of therapy) metastatic SCLC .Seizure-related homolog 6 (SEZ6) is a transmembrane protein localized to the cell surface of specific neuronal lineage cells. In murine models, the SEZ6 gene family demonstrated a regulatory influence over various aspects of neural function. Specifically, this gene family has been observed to modulate synaptic density, synaptic plasticity, and dendritic morphology within the cortical and hippocampal regions. Furthermore, SEZ6 seems to influence neuronal connectivity in the cerebellum .Interestingly, SEZ6 exhibits prominent expression within the majority of SCLC cases, as well as in other neuroendocrine and central nervous system (CNS) tumors. Conversely, SEZ6 demonstrates low expression in normal tissues and non-neuroendocrine tumor types. This expression pattern underlines SEZ6 as an attractive therapeutic target, offering the potential to maximize treatment efficacy while minimizing toxicity .ABBV-011 is an anti-SEZ6 monoclonal antibody conjugated via a non-cleavable linker drug (LD19.10) to calicheamicin, a cytotoxic payload that induces double-strand breaks in the DNA . InteresA first-in-human, phase 1 study (NCT03639194) is currently evaluating the safety and tolerability of ABBV-011 as a single agent or in combination with budigalimab  in participants with relapsed or refractory SCLC. As of the data cutoff date, August 2022, a total of 99 patients had undergone treatment with ABBV-011 monotherapy. Considering a median duration of treatment of 12 weeks (from 1.9 to 63.3 weeks), the ORR was 25%, with 10 partial responses (PRs). The median duration of response was 4.2 months , and the median PFS was 3.5 months. The clinical benefit rate (CBR) reached 65%, with 10 PRs and 16 cases of stable disease. Notably, a CBR lasting beyond 12 weeks was observed in 43% of cases. The most prevalent treatment-emergent adverse events (TEAEs) included fatigue (48%), nausea (45%), anorexia (38%), thrombocytopenia (38%), and vomiting (35%). Grade 3 TEAEs were documented in 18 patients (45%), with the most common occurrences being fatigue, thrombocytopenia, and neutropenia (each at 10%)  Table 1Table 1.In addition, a novel anti-SEZ6 ADC, ABBV-706, is currently under investigation in a phase 1 clinical trial (NCT05599984). This trial aims to evaluate the efficacy and safety of ABBV-706, both as monotherapy and in combination with budigalimab, cisplatin, or carboplatin in patients with advanced solid tumors (including SCLC) .Carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5), alternatively known as CD66e (Cluster of Differentiation 66e), belongs to the carcinoembryonic antigen (CEA) gene family, and it is involved in cell adhesion, signaling, and the promotion of cancer progression and metastasis ,49.CEACAM5 is overexpressed in numerous epithelial tumors, especially in lung cancers . HoweverTusamitamab ravtansine (SAR408701) is a targeted ADC against tumor cells expressing CEACAM5, composed of a humanized anti-CEACAM5 monoclonal antibody covalently linked to the potent cytotoxic agent, maytansinoid DM4, via a cleavable linker . The ant2 administered every 2 weeks (Q2W). In the same trial, two distinct treatment schedules were also investigated, and SAR408701 exhibited a favorable safety profile with both of these alternative dosing regimens. The MTDs were identified as 170 mg/m2 (Loading Dose), followed by 100 mg/m2 administered every 2 weeks (Q2W), and 170 mg/m2 administered every 3 weeks (Q3W) as a fixed dose [A first-in-human, phase 1 study (NCT02187848) is currently evaluating the safety and tolerability of SAR408701 in patients with advanced solid tumors, including, to date, only one patient affected by SCLC. Twenty-two patients (71%) experienced ≥1 TEAE, seven patients (22.6%) experienced ≥1 treatment-related grade ≥3 TEAE, and three patients (9.7%) discontinued treatment due to TEAEs . The mosxed dose . The development of antibody–drug conjugates (ADCs) is continuously progressing, and new potential therapies are constantly evolving, leveraging novel payloads/targets/linkers.A phase 1/2a study (NCT03221400) is currently evaluating PEN-866 in solid cancers, including SCLC. PEN-866 is an ADC that specifically targets intracellular Heat Shock Protein 90 (HSP90) and carries the topoisomerase 1 inhibitor SN-38 as a payload. PEN-866 demonstrated preliminary evidence of antitumor activity in preclinical models and in a phase I clinical trial ,54.Recently, Yamaguchi et al. demonstrated that junctional adhesion molecule 3 (JAM3) is expressed in SCLC cell lines and tissues . MoreoveAnother potential ADC target is the cell adhesion molecule neurexin-1 (NRXN1). NRXN1 is a transmembrane protein specifically overexpressed in SCLC and shows minimal to no expression in normal tissues. Using a primary anti-NRXN1 monoclonal antibody in combination with a secondary ADC demonstrated antitumor activity in SCLC cell lines .Recently, Kim et al. developed an ADC (4C9-DM1) targeting c-KIT with a microtubule inhibitor payload, which showed antitumor efficacy in SCLC cell lines and mouse xenografts .Despite the recent improvements in the development of SCLC treatments, this histological subtype continues to exhibit a poor prognosis. The introduction of immunotherapy in combination with chemotherapy has significantly improved the survival rates of patients with SCLC ,4. On thADCs, which combine the precision of monoclonal antibodies with the cytotoxic potential of chemotherapeutic agents, have already exhibited noteworthy success in treating several malignancies, including breast cancer ,59, bladOnly preliminary data are available regarding the role of ADCs in SCLC. Alongside trials with moderately promising results, such as those concerning SG , some stPatient selection is an important aspect worth exploring, particularly in identifying predictive factors that could anticipate treatment response. Despite the absence of available molecularly targeted therapies in SCLC, the application of ADCs may represent a potential treatment strategy that takes advantage of the intrinsic molecular characteristics exhibited by the tumor cells, such as the overexpression of TROP2, DLL3, B7-H3, and CECAM5.However, to date, the studies available have provided conflicting data regarding the overexpression of specific markers as predictors of treatment response. In this light, the overexpression of TROP2 has not been definitively correlated with improved therapeutic responses in patients with advanced NSCLC who have undergone anti-TROP2 ADC treatment . On the ADCs are designed to selectively deliver cytotoxic payloads specifically within the confines of neoplastic cells while sparing healthy neighboring tissues. However, studies conducted on breast cancer and NSCLC have brought to light substantial toxicities, ranging from ocular complications  to the dOverall, the novelty of ADCs means that their behavior, in terms of both efficacy and potential side effects, may not always conform to conventional expectations. Consequently, further investigations are unquestionably required to elucidate the mechanisms of action and toxicity of these specific drugs. In addition, it will be essential in the future to optimize the management of adverse events by developing ADCs with more favorable toxicity profiles and, on the other hand, by optimizing supportive therapy, potentially to prevent the most common AEs.Of note, the development of these drugs is taking place within a broader landscape of significant advancements in oncological therapeutics. Specifically, there is a growing emphasis on harnessing immune-related mechanisms to devise more effective treatments for several histologies. In this light, in a preclinical SCLC model, the combination of Rova-T with an anti-PD-1 agent showed promising results by enhancing tumor control and the immune response . SubsequWithin this rapidly evolving field, two notable and up-and-coming areas of research and development are represented by Chimeric Antigen Receptor T-cell (CAR-T cell) and bispecific T-cell engager (BiTE). CAR T-cell therapy has demonstrated remarkable success in hematological malignancies, leading to durable remissions ,67. OngoOn the other hand, bispecific antibodies have shown great promise in activating the immune response against cancer and are currently being explored in various cancer types. Of note, Tarlatamab, a bispecific T-cell engager molecule that binds to both DLL3 and CD3, initiating T-cell-mediated apoptosis, demonstrated encouraging results and manageable toxicity in ES SCLC .These developments exemplify the growing understanding of the immune system’s role in cancer control and the innovative strategies devised to exploit this knowledge.In this light, the proper role and placement of ADCs in SCLC need to be clarified, considering the forthcoming results of the ongoing trials and the emerging alternatives  in the treatment landscape.SCLC represents one of the most lethal solid cancers, requiring effective and safe therapeutic development. Within this context, ADCs emerge as a potential therapeutic alternative. The evidence in this field is steadily accumulating, with the progressive expansion of the target molecules and cytotoxic agents employed. Further studies, together with the results of ongoing trials, are eagerly awaited to better evaluate the efficacy and safety of ADCs in order to reshape the current treatment landscape of SCLC.