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Cell lines derived from circulating tumor cells (CTCs) in the blood provide important biological information on cancer metastasis. CTC-ITB-01 is a CTC cell line derived from a patient with metastatic estrogen receptor-alpha (ER-alpha) positive breast cancer two months before the death of the patient. After a LC-MC/MS based proteomics analysis of CTC-ITB-01, we found extraordinary high levels of the poorly characterized protein SUSD2 (sushi domain-containing protein 2) in CTC-ITB-01. Expression of SUSD2 on subsets of CTCs was validated on clinical blood samples of patients with metastatic breast cancer. SUSD2-positive CTCs could be captured specifically by a MACS-based approach. We overexpressed SUSD2 in the poorly-metastatic cell line MCF-7. This resulted in upregulation of ER-alpha, the tumor progression protein GRP78 (78-kDa glucose-regulated protein) and downregulation of the tumor suppressor protein PDCD4 (programmed cell death protein 4). We observed downregulation of SUSD2 and PDCD4 after hypoxia and simulation of re-oxygenation in the blood in MCF-7 and MDA-MB-468, while in CTC-ITB-01 SUSD2 levels remained unchanged, and only PDCD4 was downregulated under hypoxia. In conclusion, we show, for the first time, that SUSD2 is expressed in CTCs and appears to affect key proteins in tumor progression and survival., Competing Interests: Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

This study aimed to elucidate the potential causal relationship between 4,907 plasma proteins and the risk of gastric cancer using a two-sample Mendelian randomization approach. We utilized genome-wide association study (GWAS) data to perform two-sample Mendelian randomization analyses, treating the 4,907 plasma proteins as exposure factors and gastric cancer as the outcome. Instrumental variables for plasma proteins were selected based on strongly correlated SNPs identified through data processing and screening of the GWAS data provided by the deCode database. We employed a set of statistical methods centered on inverse variance weighting (IVW) for Mendelian randomization analysis to estimate the odds ratios (ORs) for the effects of these plasma proteins on gastric cancer susceptibility. According to the IVW method, 14 plasma proteins were associated with gastric cancer (p < 0.005). Specifically, CHST15 (OR = 0.7553, 95% CI = 0.6346 - 0.8988), L1CAM (OR = 0.7230, 95% CI = 0.5876 - 0.8896), FTMT (OR = 0.8246, 95% CI = 0.7241 - 0.9391), and PMM2 (OR = 0.5767, 95% CI = 0.3943 - 0.8433) were negatively correlated with GASTRIC CANCER, whereas ABO (OR = 1.1868, 95% CI = 1.0638 - 1.3240), FAM3D (OR = 1.2109, 95% CI = 1.0850 - 1.3515), FAM3B (OR = 1.2988, 95% CI = 1.0953 - 1.5402), ADH7 (OR = 1.3568, 95% CI = 1.1044 - 1.6670), MAP1LC3A (OR = 1.3704, 95% CI = 1.1194 - 1.6778), PGLYRP1 (OR = 1.4071, 95% CI = 1.1235 - 1.7623), PDE5A (OR = 1.7446, 95% CI = 1.2693 - 2.3978), GLUL (OR = 3.1203, 95% CI = 1.5017 - 6.4839), NFE2L1 (OR = 3.1759, 95% CI = 1.6163 - 6.2402), and MAFG (OR = 3.1945, 95% CI = 1.5329 - 6.6575) were positively correlated. Convergent results from Weighted Median and MR-Egger analyses confirmed these associations. Reverse Mendelian randomization analysis indicated that gastric cancer does not significantly alter the levels of these 14 plasma proteins (p > 0.05). Sensitivity analyses, including assessments of heterogeneity and horizontal pleiotropy, confirmed the robustness and reliability of our findings without significant bias. Pathway enrichment analysis of gene expression associated with these 14 plasma proteins, using GO and KEGG pathways, revealed that CHST15, L1CAM, FTMT, and PMM2 may serve as protective factors against gastric cancer, while ABO, FAM3D, FAM3B, ADH7, MAP1LC3A, PGLYRP1, PDE5A, GLUL, NFE2L1, and MAFG may contribute to gastric cancer pathogenesis. These results highlight the complex biological interactions between plasma proteins and tumorigenesis, providing valuable insights for preventive and therapeutic strategies in gastric malignancy management., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Glycosylation changes of circulating proteins carrying the CA19-9 antigen may offer new targets for detection methods to be explored for the diagnosis of epithelial ovarian cancer (EOC). Search for assay designs for targets initially captured by a CA19-9 antigen reactive antibody from human body fluids by probing with fluorescent nanoparticles coated with lectins or antibodies to known EOC associated proteins. CA19-9 antigens were immobilized from ascites fluids, ovarian cyst fluids or serum samples using monoclonal antibody C192 followed by probing of carrier proteins using anti-MUC16, anti-MUC1 and, anti STn antibodies and seven lectins, all separately coated on nanoparticles. Compared to reference CA19-9 and CA125 immunoassays, nanoparticle aided detection using MUC16, Ma695 and STn antibodies and lectin WGA provided, both separately and combined, improved discrimination of EOC and borderline cancers from benign samples when applied to 60 cyst fluid specimens. When applied to a panel of 44 serum samples (EOC N = 24, healthy and benign samples N = 20) two assays, CA19-9 Ma695 and CA19-9 MUC1 , stood out with equally superior separations (p-values < 10 -8 ) of the two groups compared to conventional CA19-9 immunoassay (p-value 0.03).Eu +3 -NP based CA19-9 MUC16 , CA19-9 Ma695 , CA19-9 STn and CA19-9 WGA show promise for improved EOC detection when applied to ascites & cyst fluids. When applied to circulation-derived samples, the two MUC1 based assays, CA19-9 Ma695 and CA19-9 Ma552 outperformed other assay constructs. Our results call for further validation in larger EOC cohorts preferentially with early stage ovarian cancers and all major histotypes against commonly occurring benign conditions., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics approval statement: The study was approved by the local ethics committee Regionala etikprövningsnämnden i Göteborg (Dnr 201–15) in Gothenburg and was performed in accordance with the Declaration of Helsinki and all its amendments. Patient consent statement: Informed consent was obtained from all individual participants included in the study., (© 2025. The Author(s).)

It has been documented that D-dimer levels have potential utility as a measure of tumor activity in diffuse large B-cell lymphoma (DLBCL), however whether it can be used as a predictive marker of treatment outcome has not been established. This study means to retrospectively evaluate the role of D-dimer in prediction of treatment efficacy in patients with DLBCL. 151 patients with newly diagnosed DLBCL were enrolled. Blood samples were taken from those patients during the initial visit to our hospital and again after two cycles of chemotherapy to measure D-dimer levels. The link between plasma D-dimer concentrations and patients' clinical characteristics was explored before and after treatment. D-dimer levels within the range of 0-1 µg/mL were considered negative, while readings above this range were considered positive. D-dimer difference percentage (Ddp) represents the percentage change in D-dimer levels before and after chemotherapy, calculated as (post-chemotherapy D-dimer minus pre-chemotherapy D-dimer) / pre-chemotherapy D-dimer × 100. Patients showed statistically different plasma D-dimer levels at initial consultation across treatment-response groups: CR (0.63 µg/mL [0.43-1.27]), PR (1.39 µg/mL [0.73-2.46]), SD (0.89 µg/mL [0.59-1.24]), and PD (1.34 µg/mL [0.67-2.62]). After chemotherapy, the PR group exhibited a mean D-dimer level of -0.38 µg/mL (range - 1.64 to -0.11), which was significantly lower than that of the PD group (mean 0.04 µg/mL, range - 0.40 to 0.79; p < 0.05). The CR group revealed significantly lower initial D-dimer levels (median 0.63 µg/mL) and greater reductions after chemotherapy compared to the non-CR group (median 1.17 µg/mL, p < 0.05). Patients with coagulation disorders such as DIC, DVT, or PE were excluded to minimize confounding factors. While this study demonstrates the utility of D-dimer in predicting short-term treatment response, the relationship with long-term outcomes such as PFS and OS requires further investigation. Patients who respond well to chemotherapy typically exhibit lower D-dimer levels at the initial diagnosis. Those in the SD or PD groups usually experience a greater increase in D-dimer levels following chemotherapy. Consequently, variations in plasma D-dimer levels before and after treatment may offer valuable insights for evaluating the efficacy of chemotherapy treatment., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics approval and consent to participate: This retrospective study was approved by the Ethics Committee of the Second Affiliated Hospital of Nanjing Medical University (approval number: 2023KY-13-01) and was conducted in accordance with the Declaration of Helsinki., (© 2025. The Author(s).)

Pancreatic ductal adenocarcinoma lacks suitable biomarkers for early diagnosis of disease. In gene panels developed for early diagnosis of pancreatic cancer, high AHNAK2 mRNA expression was one possible biomarker. In silico analysis of published human sample datasets (n = 177) and ex vivo analysis of human plasma samples (n = 30 PDAC with matched 30 healthy control) suggested AHNAK2 could be a diagnostic biomarker. At a plasma level of 421.47 ng/ml, AHNAK2 could potentially diagnose PDAC with a specificity and sensitivity of 83.33% and 86.67%. In vitro analysis suggests that in cell lines with diffuse cytoplasmic distribution of AHNAK2, there was colocalization of AHNAK2 with Cortactin in filipodia. This colocalization increased when cells were cultured on substrates such as Fibronectin and Collagen, as well as in hypoxia, and resulted in an augmented invasion of cancer cells. However, in cell lines with a vesicular AHNAK2 staining, such changes were not observed. Our study posits AHNAK2 as a valuable diagnostic biomarker in PDAC, now demanding prospective validation. Determination of mechanisms regulating AHNAK2 subcellular localisation may help explain its biological role., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethical approval: Ethics approval for the use of human pancreatic cancer tissue was obtained from Barts Pancreas Tissue Bank (BPTB; bartspancreastissuebank.org.uk; 2019/11/QM/HK/P/Blood and 2021/04/QM/HK/E/Blood&Tissue)., (© 2025. The Author(s).)

Blood-based predictive markers for the efficacy of immune checkpoint inhibitors (ICIs) have not yet been established. We investigated the association of the plasma level of S100A8/A9 with the efficacy of immunotherapy. We evaluated patients with unresectable stage III/IV or recurrent non-small cell lung cancer (NSCLC) who were treated with ICIs at Okayama University Hospital. The pre-treatment plasma levels of S100A8/A9 were analyzed. Eighty-one eligible patients were included (median age, 69 years). Sixty-two patients were men, 54 had adenocarcinoma, 74 had performance status (PS) 0-1, and 47 received ICIs as first-line treatment. The median time to treatment failure (TTF) for ICIs was 5.7 months, and the median overall survival (OS) was 19.6 months. The TTF and OS were worse in patients with high plasma S100A8/A9 levels (≥ 2.475 µg/mL) (median TTF: 4.3 vs. 8.5 months, p = 0.009; median OS: 15.4 vs. 38.0 months, p = 0.001). Multivariate analysis revealed that PS ≥ 2, liver metastasis, and high plasma S100A8/A9 levels were significantly associated with short TTF and OS. In conclusion, plasma S100A8/A9 level may have a limited effect on ICI therapy for NSCLC., Competing Interests: Declarations. Competing interests: Kadoaki Ohashi has received honoraria from Eli Lilly Japan K.K., Novartis Pharma K.K., Chugai Pharmaceutical Co., Ltd., AstraZeneca K.K., Kyowa Kirin Co., Ltd., and Novartis Pharma K.K; grants from Nippon Boehringer Ingelheim Co., Ltd., Chugai Pharmaceutical Co., Ltd., Eli Lilly Japan K.K., Daiichi Sankyo Co., Ltd., Amgen K.K., and Novartis Pharma K.K; and receipt of research reagents from Genentech, Inc. and Novartis Pharma K.K. outside the submitted work. Kiichiro Ninomiya has received honoraria from AstraZeneca K.K., Nippon Boehringer-Ingelheim Co., Ltd., Kyowa Kirin Co., Ltd., Eli Lilly Japan K.K., Chugai Pharmaceutical Co., Ltd., Nippon Kayaku Co., Ltd., TAIHO PHARMACEUTICAL CO., LTD., MSD K.K., Ono Pharmaceutical CO., LTD., Takeda Pharmaceutical CO., LTD., Pfizer Japan Inc., Bristol Myers Squibb K.K., Elekta K.K., Janssen Pharmaceutical K.K., Amgen K.K., and Daiichi Sankyo Co., Ltd. outside the submitted work. Eiki Ichihara received honoraria from AstraZeneca K.K., Novartis Pharma K.K., Janssen Pharmaceutical K.K., Chugai Pharmaceutical Co., Ltd., Eli Lilly Japan K.K., Pfizer Japan Inc., Bristol Myers Squibb K.K., Ono Pharmaceutical Co., Ltd., Takeda Pharmaceutical CO., LTD., and Boehringer Ingelheim; and research fund from Janssen Pharmaceutical K.K., Pfizer Japan Inc., Bristol Myers Squibb K.K., Ono Pharmaceutical Co. Ltd., and Takeda Pharmaceutical CO., LTD. outside the submitted work. Katsuyuki Kiura has received honoraria from AstraZeneca K.K., Nippon Boehringer-Ingelheim Co., Ltd., Merk Biopharma, TAIHO PHARMACEUTICAL CO., LTD., Chugai Pharmaceutical Co., Ltd., Takeda Pharmaceutical CO., LTD., Eli Lilly Japan K.K., Pfizer Japan Inc. Bristol Myers Squibb K.K., NipponKayaku Co., Ltd., Ono Pharmaceutical Co., Ltd., and MSD K.K.; research fund from Nippon Boehringer-Ingelheim Co., Ltd., Ono Pharmaceutical Co., Ltd., Novartis Pharma K.K., Takeda Pharmaceutical CO., LTD., Teijin pharma Ltd., Kyorin Pharmaceutical Co., Ltd., Shionogi Pharma Co., Ltd., Nippon Kayaku Co., Ltd., TAIHO PHARMACEUTICAL CO., LTD., and Chugai Pharmaceutical Co., Ltd.; and consulting fees from Nippon Kayaku Co., Ltd., and Nipro Corp. outside the submitted work. Katsuyuki Hotta has received honoraria from AstraZeneca K.K., Chugai Pharmaceutical Co., Ltd., Eli Lilly Japan K.K., MSD K.K., Bristol Myers Squibb K.K., Ono Pharmaceutical Co., Ltd., Nippon Boehringer-Ingelheim Co., Ltd., NipponKayaku Co., Ltd., Amgen K.K., TAIHO PHARMACEUTICAL CO., LTD., Merck Biopharma Co., Ltd.; and research fund from MSD K.K., AstraZeneca K.K., Chugai Pharmaceutical Co., Ltd., Eli Lilly Japan K.K., Bristol Myers Squibb K.K., Ono Pharmaceutical Co., Ltd., and Abbvie Inc. outside the submitted work. Tadahiro Kuribayashi, Rie Kinoshita, Go Makimoto, Toshio Kubo, Kammei Rai, Masahiro Tabata, Yoshinobu Maeda, Shinichi Toyooka, and Masakiyo Sakaguchi declare no potential conflict of interest., (© 2025. The Author(s).)

Cholangiocarcinoma (CCA) is an aggressive cancer originating from bile duct epithelial cells, with a high rate of recurrence following surgical resection. Recurrence is categorized as early linked to aggressive tumor biology than late recurrence. This study aimed to identify novel peptide mass fingerprints (PMFs) and potential biomarker panels in the serum of CCA patients with early and late recurrence using mass spectrometry. Serum samples of 81 CCA patients were analyzed using MALDI-TOF MS and LC-MS/MS, with statistical analysis correlating peptide profiles with clinical outcomes like disease-free survival (DFS) and overall survival (OS). A 365-day DFS cut-off effectively distinguished early from late recurrence, with early recurrence linked to poorer survival outcomes. The PMFs from MALDI-TOF MS differentiated recurrence types based on specific mass signatures. LC-MS/MS analysis identified 95 peptides associated with cancer progression in early recurrence and 60 in late recurrence. Distinct protein associations were found: ATR, POLA1, BLM, SP100, and PPP1R15A for early recurrence, and SERPINA1, TGFB2, SERPING1, and CAD for late recurrence, with strong interactions with chemotherapeutic drugs. This study successfully demonstrated the use of PMFs for rapid discrimination between early and late recurrence in CCA and identified potential serum peptide biomarkers to improve accuracy in recurrence classification., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Nutrition and inflammation are closely related to prognosis in breast cancer patients. However, current nutritional and inflammatory measures predict disease free survival (DFS) of breast cancer are still different, and the most predictive measures remain unknown. This study aimed to compare the predictive effects of commonly used nutritional and inflammatory measures on DFS and to improve existing nutritional or inflammatory measures in order to develop a new model that is more effective for predicting postoperative recurrence and metastasis in breast cancer patients. The clinical data of 536 female breast cancer patients with invasive ductal carcinoma who underwent surgery at Shaoxing People's Hospital from January 2012 to December 2018 were retrospectively evaluated. The predictive effects of nutritional and inflammatory indicators on DFS were evaluated. Machine learning was used to evaluate and rank laboratory indicators, select relatively important variables to modify nutritional or inflammatory indicators with the best predictive power, and evaluate their predictive role in patients' postoperative recurrence and metastasis. Among various metrics predicting DFS, the CONUT score emerged paramount with an area under the curve (AUC) of 0.667. Interestingly, the combination of the erythrocyte levels with the CONUT score (ECONUT) achieved the highest AUC (0.722). The Kaplan-Meier survival analysis showed that the group exhibiting high ECONUT scores experiencing a notably poorer DFS. ECONUT was identified as an independent risk factor for postoperative DFS (P < 0.001). The ECONUT model could provide an effective assessment tool for predicting DFS in breast cancer patients., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics approval: The study was conducted in accordance with the Declaration of Helsinki, and the protocol was approved by the Ethics Committee of Shaoxing People’s Hospital (064-Y-01). Due to the retrospective nature of the study, Ethics Committee of Shaoxing People’s Hospital waived the need of obtaining informed consent., (© 2025. The Author(s).)

Glioblastoma multiforme (GBM) is the most prevalent, treatment-resistant, and fatal form of brain malignancy. It is characterized by genetic heterogeneity, and an infiltrative nature, and GBM treatment is highly challenging. Despite multimodal therapies, clinicians lack efficient prognostic and predictive markers. Therefore, new insights into GBM management are urgently needed to increase the chance of therapeutic success. Circulating miRNAs (miRs) are important regulators of cancer progression and are potentially useful for GBM diagnosis and treatment. This study investigated how miR-29a, miR-106a, and miR-200a affect the prognosis of GBM patients. This study was conducted on 25 GBM patients and 20 healthy volunteers as a control group. The expression levels of target miRs were analyzed pre- and post-treatment using qRT-PCR and evaluated in relation to both clinical GBM criteria and the patient's survival modes. The diagnostic efficacy of target miRs was assessed using the receiver operating characteristic (ROC) curve. MiRs levels showed significant differences among the enrolled participants. All investigated miRs were significantly elevated in GBM patients with non-frontal lesions. Only miR-200a showed a significant difference in GBM patients older than 60 years with a tumor size ≥ 5 mm. Regarding miR-106a, a significant difference was detected based on the surgical strategy and use of an Eastern Cooperative Oncology Group (ECOG) performance status equal to 2. For miR-29a, a significant upregulation was detected according to the surgical strategy. All post-treatment miRs levels in GBM patients were significantly downregulated. In conclusion, circulating miRs revealed a significant role in predicting GBM patient treatment outcomes providing valuable insights for personalized therapeutic strategies., Competing Interests: Declarations. Ethical approval and consent to participate: This study was performed in line with the principles of the Declaration of Helsinki Guidelines. Approval was granted by the Research Ethics Committee (REC), Faculty of Medicine, Ain Shams University (Approval No. FWA 000017585). Written informed consent was obtained from all individual participants included in the study. Consent for publication: The authors affirm that human research participants provided informed consent for publication. All authors have read the manuscript in its final form and approved the publication. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Testicular germ cell tumour (TGCT) is a malignancy with known inherited risk factors, affecting young men. We have previously identified several hundred differentially abundant circulating RNAs in pre-diagnostic serum from TGCT cases compared to healthy controls. In this study, we performed Weighted Gene Co-expression Network Analysis (WGCNA) on mRNA and miRNA data from these samples. Central genes (hub genes) enriched functional pathways, and regulatory feature prediction were identified for all TGCT subtypes together and according to histology. The TGCT susceptibility genes TEX14, NARS2, and G3BP2, were identified as hub genes in both seminoma and non-seminoma networks. We also identified UBCA1, RCC1, FMR1, OAS3, and UBE2W as hub genes associated with TGCT. The genes OAS3 and UBE2W have previously been associated with testicular dysgenesis. Furthermore, network module analysis indicated transcription factors for oestrogen-related receptors to have a potential role during development of TGCT. The overlap between mRNA network hub genes and TGCT susceptibility genes indicates a common role in TGCT development., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethical approval: The studies involving human participants were reviewed and approved by the Norwegian Regional Committee for Medical and Health Research Ethics (REC no: 2016/1590). All methods were performed in accordance with the relevant guidelines and regulations. The patients/participants provide their written informed consent to participate in this study., (© 2025. The Author(s).)

In this study, we measured human epidermal growth factor receptor (EGFR) mutations in both tissue and circulating tumor DNA (ctDNA) by using beads, emulsions, amplifications and magnetic polymerase chain reaction (BEAMing PCR). Noninvasive mutation detection by assessing circulating tumor DNA (ctDNA) offers many advantages over tumor biopsy. One hundred non-small cell lung cancer (NSCLC) patients were enrolled, and both preoperative plasma samples and formalin-fixed and paraffin-embedded (FFPE) samples were collected for the study. The EGFR mutation status was determined by BEAMing PCR in ctDNA. Real-time quantitative PCR (qPCR) data were collected from our hospital database (EMR-qPCR, Electronic Medical Records) for comparative analysis. Additionally, qPCR was also performed on FFPE tissues using a Diatech EGFR qPCR kit. The concordance rates were 98.8%, 98.9% and 95.5% for exons 19, 20 and 21, respectively, when the BEAMing data were compared with the EMR-qPCR data. Additionally, when the BEAMing and Diatech qPCR data were compared, 90%, 100%, 96% and 98% of the genes were obtained for exons 19, 20, 21 (L858R) and 21 (L861Q), respectively. For both comparisons, Cohen's kappa agreement was significant. The advantage of BEAMing is its ability to identify mutated DNA sequences in cancer cells in the background of normal cell DNA contamination. This could be useful for disease monitoring and progression., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics declarations: This study was approved by the ACTREC-TMC institutional review board, and all participants provided informed consent., (© 2025. The Author(s).)

No biomarker can effectively screen for early gastric cancer (EGC). Players in the A disintegrin and metalloproteinase (ADAM)-natural killer group 2 member D (NKG2D) receptor axis may have a role for that. As a proof-of-concept pilot study, the expression of ADAM8, ADAM9, ADAM10, ADAM12, ADAM17, and major histocompatibility complex (MHC) class I chain-related sequence A (MICA), a ligand for NKG2D, in gastric cancer was investigated in silico using The Cancer Genome Atlas (TCGA) database. Subsequently, the mRNA and protein expression levels of these markers except ADAM8 were tested in blood samples from patients with EGC and healthy controls. In the TCGA data analyses, EGC tissues (n = 57) expressed significantly higher mRNA levels of ADAM8, ADAM9, ADAM10, ADAM12, and ADAM17 than normal tissues (n = 35) (p < 0.005). In human blood sample analyses, ADAM12 (p = 0.0007), ADAM17 mRNA (p < 0.0001) and ADAM10 (p < 0.001 7 ) protein were significantly elevated in patients with EGC (n = 27 for mRNA and n = 25 for protein) compared to the controls (n = 30 for mRNA and n = 26 for protein). Areas under the curves calculated by receiver-operating characteristic analysis for ADAM12, ADAM17 mRNA and ADAM10 protein were 0.7568 (95% confidence interval [CI]: 0.6334 to 0.8802), 0.8062 (95% CI: 0.6889 to 0.9234; p < 0.0001), and 0.8108 (95% CI: 0.6895 to 0.9320; p = 0.0001), respectively. Thus, ADAM12, ADAM17 mRNA and ADAM10 protein levels in peripheral blood could hold potential as biomarkers for screening EGC, and further investigations are required., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Microvascular invasion (MVI) diagnosis relies on postoperative pathological examinations, underscoring the urgent need for a novel diagnostic method. C-Reactive Protein (CRP), has shown significant relevance to hepatocellular carcinoma (HCC) prognosis. This study aims to explore the relationship between preoperative serum CRP levels and microvascular invasion in hepatocellular carcinoma and develop a nomogram model for predicting MVI. Patients were categorized into MVI-positive and MVI-negative groups for analysis. Serum CRP levels were compared between the two groups. And then use LASSO regression to screen variables and build a nomogram. CRP levels showed significant differences between the MVI-positive and MVI-negative groups. Multivariable logistic regression analysis identified CRP (OR = 4.85, P < 0.001), lnAFP (OR = 3.11, P < 0.001), WBC count (OR = 2.73, P = 0.003), and tumor diameter (OR = 2.38, P = 0.01) as independent predictors of MVI. A nomogram based on these variables showed good predictive performance in both the training and validation cohorts with dual validation. The clinical prediction nomogram model, which includes serum CRP levels, WBC count, tumor diameter, and serum AFP levels, showed good performance in predicting MVI in both the training and validation cohorts., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Institutional review board statement: This research complied with all principles and regulations outlined in the Declaration of Helsinki and was approved by the ethics review committee of scientific research projects of the first affiliated hospital of Zhengzhou university (2024-KY-0282-002). Informed consent: This study was a retrospective study, and the hospital ethics committee waived the patient’s informed consent., (© 2024. The Author(s).)

Integration of multi-omic data for the purposes of biomarker discovery can provide novel and robust panels across multiple biological compartments. Appropriate analytical methods are key to ensuring accurate and meaningful outputs in the multi-omic setting. Here, we extensively profile the proteome and transcriptome of patient pancreatic cyst fluid (PCF) (n = 32) and serum (n = 68), before integrating matched omic and biofluid data, to identify biomarkers of pancreatic cancer risk. Differential expression analysis, feature reduction, multi-omic data integration, unsupervised hierarchical clustering, principal component analysis, spearman correlations and leave-one-out cross-validation were performed using RStudio and CombiROC software. An 11-feature multi-omic panel in PCF [PIGR, S100A8, REG1A, LGALS3, TCN1, LCN2, PRSS8, MUC6, SNORA66, miR-216a-5p, miR-216b-5p] generated an AUC = 0.806. A 13-feature multi-omic panel in serum [SHROOM3, IGHV3-72, IGJ, IGHA1, PPBP, APOD, SFN, IGHG1, miR-197-5p, miR-6741-5p, miR-3180, miR-3180-3p, miR-6782-5p] produced an AUC = 0.824. Integration of the strongest performing biomarkers generated a 10-feature cross-biofluid multi-omic panel [S100A8, LGALS3, SNORA66, miR-216b-5p, IGHV3-72, IGJ, IGHA1, PPBP, miR-3180, miR-3180-3p] with an AUC = 0.970. Multi-omic profiling provides an abundance of potential biomarkers. Integration of data from different omic compartments, and across biofluids, produced a biomarker panel that performs with high accuracy, showing promise for the risk stratification of patients with pancreatic cystic lesions., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics approval and consent to participate: This work was performed in accordance with the Code of Ethics of the World Medical Association (Declaration of Helsinki) for experiments involving humans. Patients provided informed consent for sample and data acquisition, and the study received full ethical approval from Tallaght University Hospital Joint Research Ethics Committee Review Board (ID: 0319-264)., (© 2024. The Author(s).)

Using Fourier Transform Infrared spectroscopy (FTIR), it is possible to show chemical composition of materials and / or profile chemical changes occurring in tissues, cells, and body fluids during onset and progression of diseases. For diagnostic application, the use of blood would be the most appropriate in biospectroscopy studies since, (i) it is easily accessible and, (ii) enables frequent analyses of biochemical changes occurring in pathological states. At present, different studies have investigated potential of serum, plasma and sputum being alternative biofluids for lung cancer detection using FTIR. However, until now, it has not been shown which biofluid; among serum and plasma, that can serve as the best material medium for detecting lung cancer with highest levels of accuracy. In this study, plasma and serum isolated from blood consenting participants without lung cancer symptoms (controls) and lung cancer patients. The samples were measured using FTIR and subsequently analyzed by machine learning (ML) algorithms in order to show which fluids (serum or plasma) would better enhance detection of lung cancer. Higher absorbances values of PO 2- , CH 2 , CH 3 and amides vibrations in FTIR spectra of both serum and plasma samples, collected from lung cancer patients were observed in comparison to individuals without lung cancer symptoms (controls). Principal component analysis (PCA) of FTIR spectra showed plasma and serum samples collected from lung cancer patients and individuals without lung cancer symptoms were better differentiated in fingerprinting region (from 800 to 1800 cm - 1 ) when compared to lipid region (2800-3000 cm - 1 ). Moreover, also sensitivity specificity and accuracy calculated by logistic regression (LR) and receive operating characteristic (ROC) showed higher values for fingerprint range (800-1800 cm - 1 ) in comparison with lipids (2800-3000 cm - 1 ) one for both, serum and plasma. However, using these methods differences between serum and plasma were not existed. From the all obtained results, it was visible, that both fluids could be used in detected lung cancer using FTIR. Moreover, it was also showed that fingerprint range gave a better distinction between the studied patient groups than the lipid range. This was noticeable for both serum and plasma., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Proteases are promising biomarkers for cancer early detection. Their enzymatic activity against peptide substrates allows for their straightforward detection using low-cost tests. However, the complexity of the human proteome makes it challenging to develop sensitive and selective tests against a specific protease biomarker. Here, we report a different approach by utilizing the total protease activity in plasma samples to detect pancreatic cancer. Instead of targeting a specific protease using a specific peptide substrate, we utilized an array of 360 FRET substrates to screen for cleavage patterns in plasma samples collected from screen negatives and pancreatitis or pancreatic ductal adenocarcinoma cancer (PDAC) patients. In this proof of concept study, we first screened all 360 substrates using a small cohort (n = 13) to identify the top 5 substrates that best separate different conditions. Then, we performed a validation study using a larger cohort (n = 86) and the selected substrates. There was a statistically significant increase in the total protease activity in PDAC samples compared to screen negative and pancreatitis samples. The selected substrates detected PDAC with an area under the curve (AUC) of 0.8. This work represents a novel strategy for identifying peptide substrates for the detection of PDAC and other cancers., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Esophageal adenocarcinoma (EAC) is an aggressive cancer characterized by a high risk of relapse post-surgery. Current follow-up methods (serum carcinoembryonic antigen detection and PET-CT) lack sensitivity and reliability, necessitating a novel approach. Analyzing cell-free DNA (cfDNA) from blood plasma emerges as a promising avenue. This study aims to evaluate the cost-effective and genome-wide cell-free reduced representation bisulfite sequencing (cfRRBS) method combined with computational deconvolution for effective disease monitoring in EAC patients. cfDNA methylation profiling with cfRRBS was performed on 162 blood plasma samples from 33 EAC cancer patients and 28 blood plasma samples from 20 healthy donors. The estimated tumor fraction for EAC patients at the time of diagnosis was significantly different from the healthy donor plasma samples (one-sided Wilcoxon rank-sum test: p-value = 0.032). Tumor fractions above 15% and focal gains/amplifications in MYC (chr8), KRAS (chr12), EGFR (chr7) and NOTCH2 (chr1) were observed in four samples of distinct patients at the time metastatic disease was detected. This study showed feasibility to estimate tumor fractions in blood plasma of EAC patients based on cfDNA methylation using cfRRBS and computational deconvolution. Nevertheless, in this study only cancer patients with evidence of metastatic disease show high tumor fractions and copy number alterations., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics: Written informed consent was obtained from all cancer patients and healthy donors. Sample collection was approved by the ethics committee of Ghent University Hospital (registration numbers B670201628317, B670201628319 and B670201733701). The research was conducted according to the local legislation and institutional requirements., (© 2024. The Author(s).)

Equine melanocytic neoplasm (EMN) represents a form of skin tumor observed predominantly in grey horses aged over 15 years. Despite its prevalence, current therapeutic and preventive strategies for EMN have been subject to limited investigation. This study endeavors to shed light on potential phosphoproteins present in equine serum and fecal samples, potentially linked to EMN, with a specific focus on functional interactions in EMN pathogenesis. We examined 50 samples (25 serum, 25 feces), divided into three groups based on EMN severity: normal (n = 16), mild (n = 18), and severe EMN (n = 16). Equine phosphoproteome analysis identified 2,359 annotated serum phosphoproteins and 2002 annotated fecal phosphoproteins through differentially expressed proteins (DEPs). KEGG analysis emphasized the role of environmental information processing. Notably, the integrin NF-kappaB binding P-TEFb to stimulate transcriptional elongation signaling pathway, involving SNW1 protein, was implicated in early stage of EMN development in both serum and fecal samples. This highlights SNW1's potential role in mediating transcriptional processes, offering a novel marker within environmental information processing. This study enhances understanding of EMN mechanisms in horses, suggesting early detection through non-invasive methods and identifying a functional pathway involving SNW1, which could inform future treatment and prevention strategies., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics approval: All methods were carried out in accordance with relevant guidelines and regulation, and the study was performed in compliance with the ARRIVE guidelines. The research ethics was approved by the Faculty of Veterinary Science, Mahidol University-Institute Animal Care and Use Committee (FVS-MU-IACUC-Protocol No. MUVS-2020-12-62), Animal use license No. U1-3498-2545., (© 2024. The Author(s).)

This study aimed to investigate the correlation between baseline MRI features and baseline carcinoembryonic antigen (CEA) expression status in rectal cancer patients. A training cohort of 168 rectal cancer patients from Center 1 and an external validation cohort of 75 rectal cancer patients from Center 2 were collected. A nomogram was constructed based on the training cohort and validated using the external validation cohort to predict high baseline CEA expression in rectal cancer patients. The nomogram's discriminative ability and clinical utility were tested using the receiver operating characteristic (ROC) curve, and decision curve analysis (DCA). The baseline CEA high-expression group had significantly higher MRI-detected metastatic lymph node (mLN), MRI-detected extramural vascular invasion (mEMVI), infiltrating tumor border configuration (iTBC), peritoneal invasion, annular infiltration, maximum extramural depth (MED), and tumor length than the normal CEA group (P < 0.05). Among them, MED [odds ratio (OR):1.19 (1.03-1.38), P = 0.016] and annular infiltration [OR:2.36 (1.06-5.25), P = 0.036] were independently predicting factors for high baseline CEA expression. The trained and validated model for predicting high baseline CEA expression in the training and external validation cohorts had the area under the curves (AUC) of 0.787 (95% CI 0.716-0.859) and 0.799 (95% CI 0.698-0.899), respectively. The calibration curves of both cohorts demonstrated good agreement between predicted and observed outcomes. Decision curve analysis indicated the clinical value of the nomogram. We developed a visual nomogram to predict high baseline CEA expression for patients with rectal cancer, enabling clinicians to conduct a personalized risk assessment and therapy., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics approval and consent to participate: Due to the retrospective nature of the study, the Ethics Committee of Beijing Friendship Hospital (Approval No.2022-P2-332-01) and Tai’an City Central Hospital (Approval No. 2024-05-93) waived the need of obtaining informed consent. Consent for publication: Individual consent for this retrospective analysis was waived., (© 2024. The Author(s).)

DNA methylation is known to be involved in tumor progression. This is the first study to perform an extensive methylation analysis of plasma circulating tumor DNA (ctDNA) using targeted bisulfite sequencing in gastric cancer (GC) patients to evaluate the usefulness of ctDNA methylation as a new biomarker. Sixteen patients who received chemotherapy for recurrent GC were included. After confirmation of the methylation status of 63 genes using the Cancer Genome Atlas (TCGA) dataset, the methylation status in paired tumor and non-tumor tissues and plasma were investigated using targeted bisulfite sequencing in these genes. Forty-four of the 63 genes were significantly hypermethylated in GC patients in the TCGA cohort. Of these 44 genes, hierarchical clustering showed that five (SPG20, FBN1, SDC2, TFPI2, SEPT9) were particularly hypermethylated in tumor compared to non-tumor tissues in our GC cohort. In plasma methylation analysis, patients with high methylation of these genes had significantly worse overall survival than those with low methylation (log-rank P = 0.009). In a patient who underwent blood sampling at multiple points, the methylation levels of these five genes varied closely with clinical tumor status. The plasma ctDNA methylation levels of these five genes could be useful as a noninvasive prognostic biomarker for GC., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Platelets possess cancer-induced reprogramming properties, thereby contributing to RNA profile alterations and further cancer progression, while the former is considered a promising biosource for cancer detection. Hence, tumor-educated platelets (TEP) are considered a prospective novel method for early breast cancer (BC) screening. Our study integrated the data from 276 patients with untreated BC, 95 with benign disease controls, 214 healthy controls, and 2 who underwent mastectomy in Chinese and European cohorts to develop a 10-biomarker diagnostic model. The model demonstrated high diagnostic performance for BC in an independent test set (n = 177) with an area under the curve of 0.957. The sensitivity for BC diagnosis was 89.2%, with 100% specificity in asymptomatic controls, while that for the symptomatic group, including benign tumors and inflammatory diseases, was 62.1%. The model demonstrated substantial accuracy for stages 0-III BC (80% for stage 0 [n = 5], 83.3% for stage I [n = 12], 94.6% for stage II [n = 37], and 88.9% for stage III [n = 9]) and precisely helped determine residual cancer in two patients who underwent mastectomy. Moreover, our developed classifiers distinguish different BC subtypes properly. In summary, we created and tested a new TEP-RNA-based BC diagnostic model that was confirmed valid and demonstrated high efficiency in detecting early-stage BC and heterogeneous subtypes, including recurrent tumors. However, these results warrant more validation in larger population-based prospective studies before clinical implementation., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Study approval: This study was approved by the Ethics Committees of the First Affiliated Hospital of Xiamen University (XMYY-2021KYSB005). This study does not include any experiments with animals performed by any of the authors., (© 2024. The Author(s).)

Colorectal cancer (CRC) is one of the most common malignant tumors worldwide, with a high incidence rate and mortality. The analysis of serum biomarkers for colorectal cancer diagnosis has attracted more and more attention because of its low cost, repeatability, and quantification. This study was aimed to evaluate the diagnostic performance of serum Ephrin-A1 in patients with CRC. We retrospectively analyzed CRC cases in a test cohort (121 patients and 108 controls) and validated them in a validation cohort (119 patients and 118 controls). The concentration of Ephrin-A1 in serum was detected by Enzyme-linked immunosorbent assay (ELISA) and the diagnostic performance of serum Ephrin-A1 was evaluated by receiver operating characteristic (ROC) analysis. In the test cohort, serum Ephrin-A1 levels in patients with all-stage CRC and early-stage CRC were significantly higher than those in healthy controls. The area under the ROC curve (AUC), sensitivity and specificity of all-stage CRC and early-stage CRC were 0.709 (95% CI 0.644-0.775) and 0.660 (95% CI 0.530-0.790), 48.76% and 45.00%, 81.48% and 81.48%, respectively. Similar results were observed in the validation cohort. Serum Ephrin-A1 might be served as a potential biomarker in the diagnosis of CRC., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Breast cancer (BC) is a leading cause of global female cancer-related deaths, despite treatment advancements. A growing focus on investigating microRNA-based therapeutics and their role in BC progression. A computational analysis was performed to identify the potential miRNA-mRNA network involved in the BC pathogenesis and assist with the treatment strategy. Then, the expression levels of five circulatory miRNAs (miR-200a-3p, miR-124-3p, miR-205-5p, miR-15a-5p, and miR-155-5p) were assessed by using qRT-PCR in 75 BC patients (early-stage: n = 26 and late-stage: n = 49) and 20 healthy controls. The analysis included various (a) stages (early and late) and (b) receptor statuses (ER + ve & HER2 -ve), (HER + ve & ER -ve), and triple-negative (TNBC). In-silico analysis suggested that STAT3 serves as an efficacy biomarker suppressed by miR-124-3p. Additionally, the miR-155-5p showed the ability to activate CTNNB1 which acts as a biomarker for BC progression, to inhibit DNA repair genes (ARID2, and WEE1), and the transcriptional factor gene (TCF4). MiR-205-5p and miR-16 suppressed VEGFA expression, a survival factor for BC. MiR-200a-3p, miR-205-5p, and miR-124-3p showed downregulation in the serum of BC patients compared to controls. The ROC analysis of those miRNAs demonstrated their significant diagnostic accuracy for identifying BC patients. Additionally, miR-155-5p exhibited a significant upregulation in TNBC and can be used as an indicative marker for TNBC. This study holds significant promise for the development of noninvasive miRNA biomarkers with potential clinical applications., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Post-neoadjuvant therapy (post-NAT) and post-surgical circulating tumor DNA (ctDNA) risk stratification may enhance the management of patients with locally advanced rectal cancer (LARC). In this study, we assessed the prognostic value of ctDNA-based MRD detection in LARC patients using a personalized, tumor-informed ctDNA assay. Plasma samples from LARC patients (N = 30) were analyzed retrospectively using the Signatera™ assay. The neoadjuvant rectal (NAR) score was calculated and compared to ctDNA status to predict recurrence risk and survival outcomes. ctDNA-positive patients post-NAT and post-surgery had worse Disease Free Survival (DFS) (HR: 7.82; p = 0.001, HR: 19.65; p = 0.001) when compared to ctDNA-negative patients. In the post-NAT setting, patients who responded to NAT had superior DFS compared to patients who did not clear their ctDNA or showed no radiological response (HR: 24.7, p = 0.001 and HR: 5.1, p = 0.054, respectively). When ctDNA status is used alongside the NAR score in the post-NAT setting, patients who were ctDNA-positive with an intermediate or high NAR score showed significantly worse DFS (HR: 47.5, p < 0.001) compared to ctDNA-negative patients with either a low or intermediate/high NAR score (HR: 9.8, p = 0.0301). Post-NAT ctDNA status, whether used alone or in combination with the NAR score, may predict NAT response, and improve risk stratification., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Hepatocellular carcinoma (HCC) is often detected at advanced stages among patients with hepatitis B virus (HBV), underscoring the urgency for more precise surveillance tests. Here, we compare the clinical performance of the novel - GAAD (gender [biological sex], age, alpha-fetoprotein [AFP], protein-induced by vitamin K absence-II [PIVKA-II]) and GALAD (gender [biological sex], age, AFP, Lens-culinaris AFP [AFP-L3]), PIVKA-II) algorithms to assess the utility of AFP-L3 for distinguishing HCC from benign chronic liver disease (CLD) in Chinese patients with predominantly chronic HBV infection. Eligible adults were enrolled, and biomarkers were measured using Elecsys (Cobas) or µTASWAKO assays. In total, 411 participants provided serum samples (HCC, n = 176 [early-stage, n = 110]; CLD, n = 136; specificity n = 101). HBV was the underlying disease etiology for most participants (HCC, 95%; benign CLD, 72%). For GAAD (Cobas), GALAD (Cobas), and GALAD (µTASWAKO), AUCs were 93.1% (95% CI: 90.0-96.2), 93.2% (90.0-96.3), and 92.7% (88.4-96.9) for early-stage, and 95.6% (93.6-97.6), 95.6% (93.6-97.7), and 95.8% (93.2-98.3) for all-stage HCC, versus CLD, respectively. Interestingly, both GAAD and GALAD algorithms demonstrated comparable diagnostic performance regardless of disease etiology (HBV vs. non-HBV), presence of cirrhosis, geographic region, and within pan-tumor specificity panels (p < 0.001), indicating AFP-L3 may have a negligible role in HCC surveillance., Competing Interests: Declarations. Competing interests: HLYC reports consultancy fees from Arbutus Biopharma, Gilead Sciences, Glaxo-Smith-Kline, Roche, Vir Biotechnology, Aligos Therapeutics, Vaccitech, and Virion Therapeutics; and speaker’s bureau participation for Echosens, Gilead Sciences, Roche, and Viatris. YH has no conflicts of interest to declare. KMad, KK, and AS are employees of Roche Diagnostics International AG. KMal is an employee of Microcoat Biotechnologie, contracted by Roche Diagnostics. JH reports speaker’s bureau participation for Glaxo-Smith-Kline, Gilead Sciences, and Roche Diagnostics; has received grant/research support from Gilead Sciences and BMS; and has served on an advisory committee or review panel for Aligos, Assembly, Glaxo-Smith-Kline, Gilead Sciences, Johnson Pharmaceutica, and Roche., (© 2024. The Author(s).)

Accumulating evidence has shown that tRNA-derived small RNAs (tsRNAs) play crucial roles in malignant tumor development. However, whether serum tsRNAs can act as potential biological markers for hepatocellular carcinoma (HCC) are still largely unknown. In the current study, a novel tsRNA, namely tsRNA-Thr-5-0015, was prominently elevated in the sera of HCC patients than that of hepatitis cases and healthy check-ups, and it was related with TNM stage and lymphatic metastasis of HCC patients. Moreover, methodological evaluation confirmed that tsRNA-Thr-5-0015 had excellent stability, precision, accuracy and linear range. Additionally, the combined detection of serum tsRNA-Thr-5-0015 with AFP as well as PIVKA-II improved the diagnostic sensitivity for HCC. Furthermore, dynamic monitoring found that the serum tsRNA-Thr-5-0015 was drastically decreased in the postoperative HCC patients. Besides, Kaplan-Meier analysis displayed that patients with high level of serum tsRNA-Thr-5-0015 had shorter overall survival than that of the low level ones. In addition, bioinformatic prediction unveiled that the downstream targets of tsRNA-Thr-5-0015 were enriched in several signaling pathways, such as MAPK, PI3K-Akt, etc. In summary, tsRNA-Thr-5-0015 may be a promising biomarker for HCC diagnosis, therapeutic effect assessment and prognosis judgement. Especially, the combination with serum tsRNA-Thr-5-0015, AFP and PIVKA-II can enhance the diagnostic efficiency for HCC., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics statement: Clinical samples were collected after informed consent was obtained from the subjects, and studies involving human subjects were reviewed and approved by the Ethics Committees of the Affiliated Hospital of Nantong University (Number: 2019-L071)., (© 2024. The Author(s).)

Renal and ovarian clear cell carcinoma (CCC) are both characterized by a clear cytoplasm and exhibit similar genomic alterations and clinical characteristics. We hypothesized that both CCCs may share clinical biomarker. Tissue factor pathway inhibitor 2 (TFPI2), a serine protease inhibitor, has emerged as a promising serum biomarker for ovarian CCC, and we evaluated the efficacy of TFPI2 as a biomarker for renal cell carcinoma (RCC). Serum samples were collected from patients with RCC and healthy volunteers, and TFPI2 levels were measured. Expression of TFPI2 in each cell type was evaluated using single-cell RNA sequencing. Survival analyses according to TFPI2 expression levels were performed based on publicly available databases. Serum TFPI2 was significantly elevated in patients with RCC compared to healthy volunteers, particularly those with clear cell histology. Metastatic RCC tumors exhibited higher TFPI2 than localized RCCs. Moreover, higher TFPI2 correlated with higher Fuhrman grades in clear cell RCC. Publicly available databases showed an association between TFPI2 expression and overall survival, particularly in clear cell RCC. Single-cell RNA sequencing confirmed TFPI2 expression in clear cell RCC and normal kidney tubular epithelial cells. TFPI2 has emerged as a potential serum biomarker for RCC, offering avenues for improved detection and prognostication., Competing Interests: Competing interests The authors declare no competing interests. Ethical approval This study was performed in accordance with the Declaration of Helsinki and the Ethical Guidelines for Medical and Health Research Involving Human Subjects, after approval by the Institutional Ethics Committee of Yokohama City University (B181100031, B200800009, and B210300038). Informed consent Written informed consent was obtained from all patients for their data to be used for research purposes., (© 2024. The Author(s).)

Intestinal permeability has been related to colorectal cancer (CRC) development. Zonulin, a protein able to regulate tight junction function and intestinal permeability, emerges as a promising marker to elucidate the contribution of bacterial translocation in CRC. An Italian case-control study included 77 CRC cases, 72 intestinal adenoma and 76 healthy controls (for a total of 148 tumor-free subjects), aged 20-85. Serum zonulin levels were quantified by ELISA kit and blood 16S rRNA gene copies by DNA extraction and polymerase chain reaction. We applied logistic regression models adjusted for center, sex, age and education. There was a positive association between zonulin and CRC risk. The odds ratio (OR) of CRC for the highest versus lowest tertile of zonulin as compared to tumor-free subjects was 2.36 (95% confidence interval, 1.14-4.86). The ORs were similar in colon and rectal cancers. The OR of colon cancer for the highest versus lowest levels of both zonulin and 16S rRNA gene copies was 4.55.Circulating levels of zonulin were higher in CRC patients compared to tumor-free controls supporting the hypothesis of an interplay of gut barrier dysfunction and bacterial translocation in colorectal carcinogenesis. Zonulin may interact with 16S rRNA gene copies and serve as a further biomarker in the evaluation of CRC diagnosis., Competing Interests: Declarations Ethics approval and consent to participate The study was conducted according to the guidelines of the Declaration of Helsinki, and ap-proved by the Ethics Committees of ASST Grande Ospedale Metropolitano Niguarda (No. 477–112016; November 25, 2016) and Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico (No. 742–2017; December 14, 2017). Written informed consent was obtained from all subjects involved in the study. Consent for publication All authors consent to the publication of the present version of the manuscript. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Fluorouracil-based chemotherapy responses in colorectal cancer (CRC) patients vary widely, highlighting the role of pharmacogenomics in developing better predictive models. We analyzed 379 CRC patients receiving fluorouracil-based chemotherapy, collecting data on fluorouracil metabolism-related SNPs (TYMS, MTHFR, DPYD, RRM1), blood inflammatory markers, and clinical status. Six machine learning models-K-nearest neighbors, support vector machine, gradient boosting decision trees (GBDT), eXtreme Gradient Boosting (XGBoost), LightGBM, and random forest-were compared against multivariate logistic regression and a deep learning model (i.e., multilayer perceptron, MLP). Feature importance analysis highlighted seven predictors: histological grade, N and M staging, monocyte count, platelet-to-lymphocyte ratio, MTHFR rs1801131, and RRM1 rs11030918. In a five-fold cross-validation, XGBoost and GBDT exhibited superior performance, with Area Under Curve (AUC) of 0.88 ± 0.02. XGBoost excelled in identifying favorable prognosis (recall = 0.939). GBDT demonstrated balance in recognizing both categories, with a recall for favorable prognosis of 0.908 and a precision for unfavorable prognosis of 0.863. MLP had a similar AUC (0.87) with high precision for favorable prognosis (recall = 0.946). In external validation, XGBoost model achieved an accuracy of 0.79. An online prognostic tool based on XGBoost was developed, integrating metabolism-related SNPs and inflammatory markers, enhancing CRC treatment precision and supporting tailored chemotherapy., Competing Interests: Data Availability  The data that support the findings of this study are openly available in Github at https://github.com/xxxxljyf/SNPs-and-Blood-Inflammatory-Marker-Featured-Machine-Learning-Models-in-CRC., (© 2024. The Author(s).)

Reversing cancer cachexia remains a challenge. Genetic biomarkers for the early detection of the disease have been explored in order to enable the implementation of preventive measures. We therefore genotyped candidate genes based on cachexia phenotype and quantified adiponectin and GDF-15 levels in cachectic patients with gastrointestinal cancer. Patients with a diagnosis of gastrointestinal cancer were divided into a cachectic and a non-cachectic group after the start of chemotherapy. A control group (no cancer) was also included. We genotyped the following single nucleotide polymorphisms (SNPs) by quantitative PCR: FOXO3 (rs1935949), FOXO3 (rs4946935), ACVR2B (rs2268757), and SELP (rs6136). In addition, we quantified adiponectin and GDF-15 levels by ELISA. The rs2268757 SNP in the ACVR2B gene was associated with the weight loss phenotype in cachectic patients with gastrointestinal cancer (non-cachectic, P = 0.004). Plasma adiponectin levels were higher in cachectic patients compared to controls (P = 0.01) and non-cachectic patients (P = 0.004). GDF-15 was also elevated in cachectic patients compared to controls (P < 0.0001) and non-cachectic patients (P = 0.001). Analysis by sex showed elevated adiponectin levels in men (control, P = 0.01) and cachectic women (control, P = 0.04; non-cachectic, P = 0.01), as well as elevated GDF-15 levels in men (control, P = 0.002) and cachectic women (control, P = 0.002; non-cachectic, P = 0.007). However, there was no significant difference in the levels of these cytokines between cachectic men and women. The results suggest the rs2268757 SNP in the ACVR2B gene, adiponectin, and GDF-15 as potential biomarkers of cachexia in gastrointestinal cancer., Competing Interests: Declarations Ethics approval and consent to participate This study was approved by the Research Ethics Committee of the University of Ribeirão Preto (UNAERP) (protocol number 5.968.150), São Paulo, Brazil. Written informed consent was obtained from all participants in the study. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

In PDAC patients, ctDNA detection's prognostic significance needs validation especially in resected patients. This study investigated ctDNA kinetics in portal and peripheral blood before and after resection, and whether tissue mobilization during surgery influences ctDNA detection. In this single-center prospective cohort, portal and peripheral blood were drawn during pancreaticoduodenectomy before and after tissue mobilization, during 12 postoperative months and were associated with overall survival (OS), recurrence-free survival (RFS) and CA19-9 (secondary endpoints). Tumor mutations were identified using next-generation-sequencing and ctDNA detected by digital droplet PCR. From 2018 to 2022, 34 patients were included. The 2-year RFS and OS were 47.6%(95%CI[29.5; 63.6]) and 65.7%(95%CI[46.5; 79.4]) respectively. Intraoperatively, ctDNA detection in portal or peripheral blood was associated with worse RFS (HR[95%CI]3.26[1.26; 8.45],p = 0.010) and OS (HR[95%CI]5.46[1.65;18.01],p = 0.002). Portal vein sampling did not improve ctDNA detection. CtDNA levels were increased by 2.5-fold (p = 0.031) in peripheral blood after tissue mobilization but not significantly linked to RFS or OS. Detecting ctDNA intraoperatively was correlated with poorer RFS (HR [95% CI] 3.26 [1.26;8.45], p = 0.010) and 0S (HR [95% CI] 5.46 [1.65;18.01], p = 0.002). Portal vein sampling did not improve ctDNA detection. Tissue mobilization increases ctDNA levels. Intraoperative detection of ctDNA is associated with a worse prognosis., (© 2024. The Author(s).)

Liquid biopsy has an underexplored diagnostic potential in colorectal cancer (CRC). Sufficient quantity and quality of its elements (circulating cell-free DNA (ccfDNA), exosomes and exosomal RNA) are essential for accurate results. The present study aims to establish the optimal protocol for handling liquid biopsy samples. Samples were obtained by collecting peripheral blood from colorectal adenoma patients in CellSave tubes. Plasma was separated within six hours using differential centrifugation and aliquots stored at - 20/- 80 °C until further processing. Three methods for isolation of ccfDNA, and two combinations of kits for isolation of exosomes and exosomal RNA were tested. The quality and quantity of ccfDNA isolates were evaluated. Exosomes were characterised by determining size, concentration, and total and specific protein content. Expression of chosen microRNAs, miR-19a-3p and miR-92-3p, which have been implicated in CRC progression, were determined. The vacuum-column-based kit showed the highest quantities of isolated ccfDNA (P-value < 0.001). Kits for exosome isolation significantly differed in size (P-value = 0.016), concentration (P-value = 0.016) and protein content (P-value = 0.016). There was no significant difference in expressions of miR-19a-3p (P-value = 0.219) and miR-92a-3p (P-value = 0.094) between the two isolation kits. The new, adapted protocol described, enables simultaneous analysis of multiple elements when investigating potential biomarkers of CRC., (© 2024. The Author(s).)

This study aimed to measure relative telomere length (RTL) in blood leukocytes of advanced-stage NSCLC patients either with or without Osimertinib-induced ADRs and determine whether RTL could serve as a biomarker of Osimertinib-induced ADRs. Blood leukocytes RTL were measured in 63 advanced-stage NSCLC patients and 62 age-matched healthy controls using real-time polymerase chain reaction. In patients with advanced-stage NSCLC, RTL was significantly shorter than that in healthy controls (P < 0.001). Compared to patients without ADRs and those with mild/moderate ADRs, patients with severe ADRs exhibited significantly decreased RTL (P < 0.001, P < 0.001, respectively). ROC curve analysis uncovered a diagnostic value of RTL as a biomarker of Osimertinib-induced ADRs (AUC = 1.000, P < 0.001). Kaplan-Meier analysis revealed a significant association between shorter RTL and increased cumulative incidence of Osimertinib-induced ADRs in patients with advanced-stage NSCLC (P < 0.001). Shorter RTL in blood leukocytes would reflect the occurrence of Osimertinib-induced ADRs and might emerge as a promising biomarker for identifying advanced-stage NSCLC patients who are at risk of experiencing Osimertinib-induced ADRs, particularly those with severe ADRs., (© 2024. The Author(s).)

This study aimed to explore the association between inflammation-based prognostic markers and outcomes in non-small cell lung cancer (NSCLC) patients undergoing therapy with immune checkpoint inhibitors (ICIs). We conducted a comprehensive search of the Embase, PubMed, and Cochrane databases for studies that reported on the impact of inflammation-based prognostic factors, such as C-reactive protein (CRP), on the prognosis of NSCLC patients treated with ICIs. The primary outcomes of interest were overall survival (OS) and progression-free survival (PFS). Statistical analyses were performed using Stata 14 software, with assessments of publication bias and heterogeneity conducted as necessary. Our meta-analysis included 27 studies encompassing 5,174 patients, evaluating factors such as CRP, the modified Glasgow Prognostic Score (mGPS), and the CRP-albumin ratio (CAR). The analysis revealed that elevated levels of CRP were significantly correlated with both reduced PFS (I 2  = 0%, P = 0.72; HR = 1.50, 95%CI: 1.33-1.67, P < 0.01) and shorter OS (I 2  = 0%, P = 0.55; HR = 1.90, 95%CI: 1.50-2.30, P < 0.01). Similarly, elevated levels of mGPS values were associated with worse PFS (I 2  = 0%, P = 0.75; HR = 1.28, 95%CI: 1.10-1.46, P < 0.05) and OS (I 2  = 0%, P = 0.94; HR = 1.56, 95%CI: 1.31-1.81, P < 0.05). However, the relationship between elevated levels of CAR and worse outcomes for PFS (I 2  = 59.6%, P = 0.94; HR = 1.42, 95%CI: 0.74-2.11, P > 0.05) and OS (I 2  = 45.3%, P = 0.16; HR = 1.41, 95%CI: 0.70-2.13, P > 0.05) was not statistically significant. Our findings suggest that CRP and mGPS may serve as potential prognostic markers in NSCLC patients receiving immunotherapy. Nonetheless, further research with more homogeneous study populations is necessary to valid these observations., (© 2024. The Author(s).)

Transcriptomic profiling of blood immune cells offers a promising alternative to invasive, sampling bias-prone tissue-based biomarker assays for predicting immune checkpoint inhibitor (ICI) therapy response in non-small cell lung cancer (NSCLC) patients. However, the optimal analytical approach to identify systemic correlates of response still needs to be explored. We collected peripheral blood mononuclear cells and whole blood (WB) samples from 33 ICI-treated NSCLC patients before ICI treatment and at the first response evaluation. After bulk polyadenylated RNA-sequencing, we assessed differences in gene expression profiles between non-responders and responders using differential expression analysis, single sample gene set enrichment analysis (ssGSEA), and cell type deconvolution. We evaluated gene expression values, ssGSEA scores, and deconvolved cell type proportions to distinguish non-responders from responders via ROC curve (AUC) analysis, training a logistic regression classification model. Gene expression values and deconvolved proportions yielded the best results with WB samples after treatment (AUC = 0.87 and 0.85, respectively). Overall, ssGSEA scores showed superior classification performance across all sample types and timepoints (AUC > 0.7). In conclusion, transcriptomic analysis through ssGSEA demonstrated the best performance as a non-invasive biomarker for predicting clinical benefit in ICI-treated NSCLC patients, with gene expression and deconvolution on post-treatment WB samples also showing promising results., (© 2024. The Author(s).)

Thrombospondin 2 (TSP2) plays a vital role in collagen/fibrin formation, bone growth, vascular density regulation, hemostasis, and cell adhesion. Close associations of serum TSP2 with histological severity in non-alcoholic fatty liver disease and chronic hepatitis C were reported. The present study investigated the significance of circulating TSP2 in chronic hepatitis B patients. Eighty-seven biopsy-proven chronic hepatitis B patients were analyzed in cross-sectional Study 1 to search for correlations between serum TSP2 levels prior to liver biopsy and clinicopathological parameters. In longitudinal Study 2, 51 chronic hepatitis B patients with long-term follow-up (mean: 7.5 years) were examined for changes in serum TSP2 levels during nucleos(t)ide analog (NA) therapy along with trends in hepatocarciongenesis. In Study 1, serum TSP2 levels were not significantly associated with portal inflammation or fibrosis. Study 2 revealed that serum TSP2 was significantly decreased after 48 weeks of NA therapy (P < 0.001). Notably, TSP2 levels at 48 weeks of NA administration (TSP2-48W) were significantly higher in the hepatocellular carcinoma (HCC) (+) group than in the HCC (-) group (P = 0.043). Kaplan-Meier analysis showed that higher TSP2-48W (≥ 24 ng/mL) was associated with future HCC development (P = 0.030). Serum TSP2 levels may be a potential predictor of HCC development in hepatitis B patients receiving NA therapy. Longitudinal prospective studies are necessary to validate our findings., (© 2024. The Author(s).)

Cardia gastric cancer (CGC) is prevalent in East Asia, and noninvasive, cost-effective screening methods are needed. This study investigated the diagnostic value of serum pepsinogen (PG), gastrin-17 (G-17), Helicobacter pylori (H. pylori) antibodies, and proteomic profiling for CGC and precancerous lesions. We conducted a case-control study involving biopsy-confirmed patients with CGC (n = 60), low-grade intraepithelial neoplasia (CLGD, n = 60), high-grade intraepithelial neoplasia (CHGD, n = 64), and healthy controls (n = 120) matched for age and sex from high-incidence areas in China. Serological markers including PGI, PGII, G-17, and H. pylori were measured using ELISA and Western blot, while plasma protein markers were assessed using Olink® technology. The VSOLassoBag algorithm and nine machine learning (ML) algorithms were employed to identify crucial features and construct predictive models. Various evaluation metrics, including the area under the receiver-operating-characteristic curve (AUC), were utilized to compare predictive performance. Elevated PGII levels, decreased PGR, and H. pylori infection were significantly associated with an increased risk of CGC and precancerous lesions (P for trend < 0.05). The eXtreme Gradient Boosting (XGBoost) model performed best in discriminative ability among the 9 ML models. Following feature reduction based on predictive performance, a final explainable XGBoost model was developed, incorporating five protein biomarkers (CDHR2, ICAM4, PTPRM, CDC27, and FLT1). This model exhibited excellent performance in distinguishing individuals with CGC and precancerous lesions from healthy controls (AUC = 0.931 for CGC, 0.867 for CHGD, and 0.763 for CLGD), surpassing the traditional serological marker-based model. This study underscores the diagnostic potential of serological markers and proteomic profiling in the detection of CGC. Further validation and exploration of combined biomarker approaches are warranted to enhance early diagnosis and improve outcomes in high-risk populations., (© 2024. The Author(s).)

Through their involvement in cancer metabolism, alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), γ-glutamyltransferase (GGT) and lactate dehydrogenase (LDH) reflect the tumor burden and thus could have a prognostic potential for patients treated with immune checkpoint inhibitors (CPI). Therefore, this study investigated the prognostic potential of these parameters in a real-world cohort of patients with metastatic renal cell cancer (mRCC) under first-line CPI-based therapy. The retrospective study cohort included 82 mRCC patients treated with CPI-based first-line therapy between 2019 and 2023. Progression-free survival (PFS), overall survival (OS) and response rates were evaluated according to baseline levels and early dynamic changes of ALAT, ASAT, GGT and LDH. Multivariate Cox proportional hazard regression models were generated to identify independent prognosticators for PFS and OS. High baseline levels and non-normalized kinetics of ALAT, ASAT, GGT and LDH were significantly associated with shorter PFS and OS (p < 0.05), which was also reflected by lower response rates. Combining the four parameters at baseline into a 4-Risk-Score resulted in an enhanced prognostic power, as indicated by a higher C-index of 0.693 for OS compared to the individual parameters (≤ 0.663). Patients with all four risk factors present showed the worst PFS and OS. Overall, baseline levels and early kinetics of the four parameters as well as the 4-Risk-Score were identified as independent prognosticators for PFS and OS by multivariate analysis. As standard laboratory parameters, ALAT, ASAT, GGT and LDH are cost-effective and could be easily used either alone or in combination for therapy monitoring of CPI-treated mRCC patients., (© 2024. The Author(s).)

This retrospective study evaluated the use of laminin γ2 monomer (LG2m) as a predictive biomarker for hepatocellular carcinoma (HCC) in patients with chronic hepatitis B virus (HBV) infection. Serum LG2m levels were measured in two cohorts of patients: cohort 1 comprised 56 patients with chronic liver disease for assessing LG2m stability, whereas cohort 2 included 89 patients with chronic HBV infection who did not have HCC for evaluating the usefulness of LG2m measurement in HCC prediction. LG2m was highly stable in cryopreserved serum, and an increased LG2m level was significantly associated with a higher risk of HCC in chronically HBV-infected patients (P = 0.012). Multivariable Cox regression analysis revealed that high LG2m was an independent significant risk factor for HCC (hazard ratio, 7.16; 95% confidence interval, 1.31-39.2; P = 0.023). These findings suggest that LG2m may serve as a useful biomarker for the prediction of future HCC in patients with chronic HBV infection., (© 2024. The Author(s).)

Enhancing early-stage breast cancer detection requires integrating additional screening methods with current diagnostic imaging. Omics screening, using easily collectible serum samples, could serve as an initial step. Alongside biomarker identification capabilities, omics analysis allows for a comprehensive analysis of prevalent histological types-DCIS and IDC. Employing metabolomics, metallomics, and machine learning, could yield accurate screening models with valuable insights into organism responses. Serum samples of confirmed breast cancer patients were utilized to analyze metabolite and metal ion profiles, using two distinct analysis methods, proton NMR for metabolomics and ICP-OES for metallomics. The resulting responses were then subjected to discriminant analysis, progression biomarker exploration, examination of correlations between patients' metabolites and metal ions, and the impact of age and menopause status. Measured NMR spectra and metabolite relative integrals were used to achieve statistically significant discrimination through MVA between breast cancer and control groups. The analysis identified 24 metabolites and 4 metal ions crucial for discrimination. Furthermore, four metabolites were associated with disease progression. Additionally, there were important correlations and relationships between metabolite relative integrals, metal ion concentrations, and age/menopausal status subgroups. Quantified relative integrals allowed for discrimination between studied subgroups, validated with a holdout set. Feature importance and statistical analysis for metabolomics and metallomics extracted a set of common entities which in combination provides valuable insights into ongoing molecular disturbances and disease progression., (© 2024. The Author(s).)

Selective and sensitive sarcosine detection is crucial due to its recent endorsement as a prostate cancer (PCa) biomarker in clinical diagnosis. The reduced graphene oxide-cobalt nickel tungsten boron oxides (CoNiWBO/rGO) nanocomposite is developed as a non-enzymatic electrochemical sensor for sarcosine detection in PCa patients' serum. CoNiWBO/rGO is synthesized by the chemical reduction method via a one-pot reduction method followed by calcination at 500 °C under a nitrogen environment for 2 h and characterized by UV-Vis, XRD, TGA, and SEM. CoNiWBO/rGO is then deposited on a glassy carbon electrode, and sarcosine sensing parameters are optimized, including concentration and pH. This non-enzymatic sensor is employed to directly determine sarcosine in serum samples. Differential pulse voltammetry (DPV) and linear sweep voltammetry (LSV) are employed to monitor the electrochemical behavior where sarcosine binding leads to oxidation. Chronoamperometric studies show the stability of the developed sensor. The results demonstrate a wide linear range from 0.1 to 50 µM and low limits of detection, i.e., 0.04 µM and 0.07 µM using DPV and LSV respectivel. Moreover, the calculated recovery of sarcosine in human serum of prostate cancer patients is 78-96%. The developed electrochemical sensor for sarcosine detection can have potential applications in clinical diagnosis., (© 2024. The Author(s).)

Analysis of serial liquid biopsy (LB) samples has been found to be a promising approach for the monitoring of tumor dynamics in the course of therapy for patients with colorectal cancer (CRC). Currently, somatic mutations are used for tracing the dynamics of the tumor via LB. However, the analysis of the dynamic changes in the molecular signatures such as microsatellite instability (MSI) is not currently used. We hypothesized that changes in blood MSI burden (bMSI) could be registered using serial LB sampling in the course of immune checkpoint inhibitors (ICI), and that its changes could potentially correlate with treatment outcomes. We report the preliminary findings of the observational trial launched to study (NCT06414304) the dynamics of bMSI in 9 MSI-positive CRC patients receiving ICI. NGS-based MSI testing was performed on both pre-treatment FFPE and serial LB samples. For patients who had detectable bMSI burden in any of the LB samples (n = 8, 89%), median bMSI was 1.4% (range, 0.01-40%). Among patients with detectable MSI in available FFPE samples, median MSI burden was 29.3% (range, 10-40%). bMSI detected in baseline LB and FFPE samples were positively correlated (Pearson's R 0.47). Maximal variant allele frequencies of driver mutations observed in LB were also positively correlated with bMSI burden (Pearson's R 0.7). Patients who had clinical benefit had undetectable bMSI burden at follow-up. Our results provide the rationale for further validation of bMSI as a predictive biomarker of ICI in MSI-positive patients., (© 2024. The Author(s).)

The objective of this study was to investigate the correlation between Rab10 (GTP binding protein RAB10), TLR4 (Toll-like receptor 4), and NF-κB (nuclear factor kappa-B) levels and therapeutic effects in peripheral blood of patients with breast cancer after surgery. The study included 160 patients with stage I-III breast cancer who underwent surgical treatment at our hospital's Department of Breast Surgery and Oncology between January 2021 and June 2021. ELISA was used to assess Rab10, TLR4, and NF-κB levels in peripheral blood. Based on their levels of Rab10, TLR4, and NF-κB in peripheral blood, participants were categorized into two groups: the low marker expression group (72 participants with relatively low expression of Rab10, TLR4, and NF-κB: Rab10<2.0ng/ml; TLR4<2.75ng/ml; NF-κB<3.5ng/ml) and the high marker expression group (88 participants with relatively high expression: Rab10 ≥ 2.0 ng/ml; TLR4 ≥ 2.75ng/ml; NF-κB ≥ 3.5ng/ml). All participants provided informed consent to participate the study. The baseline data of the two groups of patients, the presence or absence of lymph node metastasis and recurrence within 3 years after surgery, as well as the survival status within 3 years after surgery (including median overall survival and median progression-free survival) were statistically analyzed. The expressions of Rab10, TLR4, and NF-κB in the peripheral blood of patients were detected through enzyme-linked immunosorbent assay (ELISA). Kendall's tau-b correlation analysis was conducted to examine the relationship between the expressions of Rab10, TLR4, and NF-κB and the therapeutic effects outcomes. The levels of Rab10, TLR4, and NF - κ B in peripheral blood of the high marker expression group were higher than those of the low marker expression group (Rab10: 1.87 ± 0.18 vs. 3.15 ± 0.24 ng/ml; TLR4: 2.17 ± 0.20 vs. 3.26 ± 0.25 ng/ml); NF-κB: 2.68 ± 0.27 vs. 4.63 ± 0.30 ng/ml; P < 0.05). Analyzing the relationship between patient staging and Rab10, TLR4, and NF - κ B expression, the number of patients in high marker expression group III-IV increased compared to the low marker expression group (54.55% vs. 36.12%; P < 0.05), while the number of patients in high marker expression group I-II decreased compared to the low marker expression group (45.45% vs. 63.88%; P < 0.05). It was found that the number of patients with no recurrence or metastasis in the high marker expression group decreased compared to the low marker expression group (56.81% vs. 73.61%; P < 0.05), while the number of patients with recurrence or metastasis in the high marker expression group increased compared to the low marker expression group (43.19% vs. 26.39%; P < 0.05). The median overall survival and median progression free survival in the high marker expression group were shorter than those in the low marker expression group (median overall survival: 21.45 ± 2.68 months vs. 28.38 ± 3.44 months; median progression free survival: 15.25 ± 2.37 vs. 20.72 ± 2.58 months; P < 0.05). Kendall's tau-b correlation indicated a positive correlation between the expressions of Rab10, TLR4, and NF-κB and a poor therapeutic effects (P < 0.05), suggesting that elevated levels of Rab10, TLR4, and NF-κB may lead to a worsened therapeutic effects. There is a significant correlation between the presence of Rab10, TLR4, and NF-κB in the peripheral blood of breast cancer patients. Elevated levels of Rab10, TLR4, and NF-κB are linked to an increased risk of recurrence, metastasis, reduced overall survival, and progression-free survival., (© 2024. The Author(s).)

Circulating tumour cells (CTCs), derived from primary tumours, play a pivotal role in cancer metastasis by migrating into the peripheral bloodstream. These cells are paramount in clinical research, serving as early diagnostic markers for metastatic cancer. Analysing CTC counts and their biomarker characteristics can provide invaluable insights into tumour identification, profiling, and metastatic capabilities. However, the rarity and diverse nature of CTCs in the bloodstream present significant challenges to their isolation and detection, especially in the initial stages of metastasis. Recent advancements in nanotechnology have led to the development of innovative CTC separation and detection methods. This review focuses on applying nanoparticles, nanomaterials, and microfluidic platforms to simplify the isolation and detection of CTCs. The infusion of nanotechnology in this field marks a crucial turning point, enabling the necessary progress to advance CTC research., (© 2024. The Author(s).)

Circulating genetically abnormal cells (CACs) have emerged as a promising biomarker for the early diagnosis of lung cancer, particularly in patients with pulmonary nodules. However, their performance may be suboptimal in certain patient populations. This study aimed to refine patient selection to improve the detection of CACs in pulmonary nodules. A retrospective analysis was conducted on 241 patients with pulmonary nodules who had undergone pathological diagnosis through surgical tissue specimens. Utilizing consensus clustering analysis, the patients were categorized into three distinct clusters. Cluster 1 was characterized by older age, larger nodule size, and a higher prevalence of hypertension and diabetes. Notably, the diagnostic efficacy of CACs in Cluster 1 surpassed that of the overall patient population (AUC: 0.855 vs. 0.689, P = 0.044). Moreover, for Cluster 1, an integrated diagnostic model was developed, incorporating CACs, sex, maximum nodule type, and maximum nodule size, resulting in a further improved AUC of 0.925 (95% CI 0.846-1.000). In conclusion, our study demonstrates that CACs detection shows better diagnostic performance in aiding the differentiation between benign and malignant nodules in older patients with larger pulmonary nodules and comorbidities such as diabetes and hypertension. Further research and validation are needed to explore how to better integrate CACs detection into clinical practice., (© 2024. The Author(s).)

Patients diagnosed with early-stage cancers have a substantially higher chance of survival than those with late-stage diseases. However, the option for early cancer screening is limited, with most cancer types lacking an effective screening tool. Here we report a miRNA-based blood test for multi-cancer early detection based on examination of serum microRNA microarray data from cancer patients and controls. First, a large multi-cancer training set that included 1,408 patients across 7 cancer types and 1,408 age- and gender-matched non-cancer controls was used to develop a 4-microRNA diagnostic model using 10-fold cross-validation. In three independent validation sets comprising a total of 4,875 cancer patients across 13 cancer types and 3,722 non-cancer participants, the 4-microRNA model achieved greater than 90% sensitivity for 9 cancer types (lung, biliary tract, bladder, colorectal, esophageal, gastric, glioma, pancreatic, and prostate cancers) and 75-84% sensitivity for 3 cancer types (sarcoma, liver, and ovarian cancer), while maintaining greater than 99% specificity. The sensitivity remained to be > 99% for patients with stage 1 lung cancer. Our study provided novel evidence to support the development of an inexpensive and accurate miRNA-based blood test for multi-cancer early detection., (© 2024. The Author(s).)

Docetaxel is commonly used for treatment of castration-resistant prostate cancer. Unfortunately, many prostate cancer patients develop resistance to docetaxel. Clinical markers less invasive than biopsies, such as blood samples, would be ideal for monitoring and predicting patient treatment outcomes to docetaxel. Lipid alterations are often associated with the progression of many cancers, including prostate cancer. This study investigated the use of lipids from whole blood as clinical markers for docetaxel resistance in a small cohort of patients with prostate cancer. Qualitative lipidomics was performed by liquid chromatography-tandem mass spectrometry to assess the lipid composition of prostate cancer cells exposed to docetaxel as well as whole blood from prostate cancer patients before, during and after docetaxel treatment. Three patients had castration resistant prostate cancer, three had castration sensitive prostate cancer, and four had de novo prostate cancer during the extent of the study. Mean decrease accuracy and classical univariate receiving operating characteristic curve analyses were performed to identify potential biomarkers. In total, 245 and 221 altered lipids were identified from a second stage of mass spectrometry analysis of prostate cancer cells and clinical blood samples, respectively. Both models indicated that docetaxel treatment altered ether-linked phosphatidylcholines, lysophosphatidylcholine, diacylglycerols, ceramides, hexosylceramides, and sphingomyelins. The results also indicated several lipid changes were associated with sphingolipid signaling and metabolism, and glycerophospholipid metabolism. Collectively, these data suggest the potential usage of identified lipid species as indicators of docetaxel resistance in prostate cancer., (© 2024. The Author(s).)

The assessment of ctDNA has emerged as a minimally invasive avenue for molecular diagnosis and real-time tracking of tumor progression in NSCLC. However, the evaluation of ctDNA by amplicon-based NGS has been not endorsed by all the healthcare systems and remains to be fully integrated into clinical routine practice. To compare tissue single-gene with plasma multiplexed testing, we retrospectively evaluated 120 plasma samples from 12 consecutive patients with advanced non-squamous NSCLC who were part of a prospective study enrolling treatment-naïve patients and in which tissue samples were evaluated using a single-gene testing approach. While the plasma ctDNA detection of EGFR and BRAF mutations had an acceptable level of concordance with the archival tissue (85%), discordance was seen in all the patients in whom ALK alterations were only detected in tissue samples. Among six responders and six non-responders, early ctDNA mutant allelic frequency (MAF) reduction seemed to predict radiologic responses and longer survival, whereas increasing MAF values with the emergence of co-mutations like BRAF V600E , KRAS G12V or TP53 M237I seemed to be an early indicator of molecular and radiologic progression. This report using an amplicon-based NGS assay on ctDNA underscores the real-life need for plasma and tissue genotyping as complementary tools in the diagnostic and therapeutic decision-making process., (© 2024. The Author(s).)

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies, presenting a persisting global health burden. Neutrophils have a double-edged role in tumor progression exhibiting both pro-tumor and anti-tumor functions. CD71, also known as transferrin receptor 1, performs a critical role in cellular iron uptake and is highly expressed on proliferating cells, and especially on activated immune cells. CD71 is known to be elevated in various types of solid cancers and is associated with poor prognosis, however, the expression of CD71 on neutrophils in PDAC and its potential clinical impact is still unknown. Therefore, we analyzed CD71 on circulating neutrophils in PDAC and clinical control patients and found a significant increased expression in PDAC patients. High expression of CD71 on neutrophils in PDAC patients was associated with reduced outcome compared to low expression. CD71 on neutrophils correlated positively with the levels of proinflammatory cytokines IL-6, IFN-γ, and growth factor ligands CD40-L, and BAFF in plasma of PDAC patients. Finally, we have demonstrated that high expression of CD71 on neutrophils was also associated with an increased expression of CD39 and CD25 on circulating T-cells. Based on our findings, we hypothesize that CD71 on neutrophils is associated with tumor progression in PDAC. Further studies are required to investigate the distinct functionality of CD71 expressing neutrophils and their potential clinical application., (© 2024. The Author(s).)

Despite widespread cervical cancer (CC) screening programs, low participation has led to high morbidity and mortality rates, especially in developing countries. Because early-stage CC often has no symptoms, a non-invasive and convenient diagnostic method is needed to improve disease detection. In this study, we developed a new approach for differentiating both CC and cervical intraepithelial neoplasia (CIN)2/3, a precancerous lesion, from healthy individuals by exploring CC fatty acid metabolic reprogramming. Analysis of public datasets suggested that various fatty acid metabolizing enzymes were expressed at higher levels in CC tissues than in normal tissues. Correspondingly, 11 free fatty acids (FFAs) showed significantly different serum levels in CC patient samples compared with healthy donor samples. Nine of these 11 FFAs also displayed significant alterations in CIN2/3 patients. We then generated diagnostic models using combinations of these FFAs, with the optimal model including stearic and dihomo-γ-linolenic acids. Receiver operating characteristic curve analyses suggested that this diagnostic model could detect CC and CIN2/3 more accurately than using serum squamous cell carcinoma antigen level. In addition, the diagnostic model using FFAs was able to detect patients regardless of clinical stage or histological type. Overall, the serum FFA diagnostic model developed in this study could be a powerful new tool for the non-invasive early detection of CC and CIN2/3., (© 2024. The Author(s).)