Potential Diagnostic Utility Research Articles

The potential serum sphingolipid biomarkers for distinguishing Wilson disease

BackgroundThe diagnosis of Wilson's disease (WD) remains a challenging endeavor in clinical practice. Serum sphingolipids play a significant role in the development of liver disease. In this study, we examined the serum sphingolipid profile in patients with WD and explored the potential diagnostic utility of serum sphingolipid metabolites. These metabolites may aid in distinguishing WD patients from healthy controls and identifying those with a risk of cirrhosis. MethodsThis study consecutively enrolled 26 WD patients and 88 healthy controls. We utilized high-performance liquid chromatography–tandem mass spectrometry to analyze a panel of 88 serum sphingolipid metabolites. The data were analyzed by multivariate statistical methods. ResultsAmong the 88 sphingolipids metabolites analyzed, 17 sphingolipids were observed significant differences between WD and HC groups (all P < 0.05). Notably, five sphingolipids, namely S1P (d18:1), Cer (d18:2/21:0), SM41:2, sph(d18:1), and Cer (d18:2/22:0), each with an AUC exceeding 0.9, emerged as potential biomarkers for WD. Additionally, in the comparison between WD patients with and without cirrhosis, 24 sphingolipid metabolites exhibited significant differences (all P < 0.05). We identified Cer(d18:1/20:0), Cer(d18:2/22:0), Cer(d18:2/24:0), Cer(d18:2/20:0), and Cer(d18:2/18:0), each with an AUC exceeding 0.9, as potential serological markers for WD patients with cirrhosis. ConclusionFor enhanced clinical applicability, we propose considering Cer (d18:2/22:0) as a predictive marker applicable to both WD patients and their susceptibility to cirrhosis. This particular ceramide has exhibited strong diagnostic and predictive performance. These findings have the potential to facilitate non-invasive WD diagnosis.

Expression of NKX2.2 in Non-Ewing Tumors With Round Cell Morphology.

Background Round cell sarcomas pose diagnostic challenges due to overlapping histopathological features, necessitating precise immunohistochemical markers for accurate categorization. NKX2.2 has emerged as a sensitive diagnostic tool, particularly in Ewing sarcoma. This study extends this understanding to various round-cell sarcomas, shedding light on the potential diagnostic utility of NKX2.2 beyond its established role. The nuanced exploration of NKX2.2 expression aims to enhance diagnostic strategies, prognostic assessments, and therapeutic developments in the landscape of sarcoma research. Methodology Cases were retrieved from the surgical pathology and consultation files of Shaukat Khanum Memorial Cancer Hospital and Research Center, Lahore, Pakistan. Representative hematoxylin and eosin-stained slides of six different types of already confirmed tumors, including lymphoblastic lymphoma, neuroblastoma, rhabdomyosarcoma, synovial sarcoma, Wilms tumor, and Ewing sarcoma, were reviewed by a panel of pathologists. Immunohistochemistry, utilizing a rabbit anti-NKX2.2 monoclonal antibody, was performed on formalin-fixed paraffin-embedded tissue sections. The presence of NKX2.2 was defined as moderate or high nuclear immunoreactivity in at least 5% of cells. Results The histopathological examination revealed characteristic features in each sarcoma subtype, aligning with established diagnostic criteria. In Lymphoblastic lymphoma, T-cell lineage was confirmed through TdT expression, while the atypical finding of focal NKX 2.2 expression hinted at genetic diversity. Neuroblastoma exhibited the expected salt and pepper chromatin pattern, with NKX 2.2 expression raising questions about its prognostic significance. Rhabdomyosarcoma presented primitive cells expressing desmin, and NKX 2.2 focal expression echoed previous subtype-associated studies. Synovial sarcoma displayed both monophasic and biphasic growth patterns and TLE1 expression, with NKX 2.2 variation suggesting tumor heterogeneity. In Wilms tumor, the characteristic WT1 expression was observed, while NKX2.2's absence reaffirmed its irrelevance in this context. Ewing sarcoma displayed the anticipated homogenous cell population, strong NKX2.2 expression, and CD99 positivity across various sites. Furthermore, age and gender impact on this range of sarcomas found no significant relation with an expression of NKX2.2. Conclusion In conclusion, the diverse expression profiles of diagnostic markers discovered in this study, particularly the atypical expression of NKX2.2 beyond its established role in Ewing sarcoma, signify a significant advancement. This unique finding accentuates the potential diagnostic importance of NKX2.2 in various sarcomas, presenting a novel dimension to our understanding of these malignancies.

Robust and fast stochastic 4D Flow Vector-field signature technique for quantifying composite flow dynamics from 4D Flow MRI: Application to left atrial flow in atrial fibrillation

4D flow MRI is an emerging imaging modality that maps voxel-wise blood flow information as velocity vector fields that is acquired in 7-dimensional image volumes (3 spatial dimensions + 3 velocity directions + time). Blood flow in the cardiovascular system is often complex and composite involving multiple flow dynamics and patterns (e.g., vortex flow, jets, stagnating flow) that occur and interact simultaneously. The spectrum of such complex flow dynamics is embedded in the velocity vector field dynamics derived from 4D Flow MRI. However, current flow metrics cannot fully measure high-dimensional vector-field data and embedded complex composite flow data. Instead, these methods need to break down the vector-field data into secondary scalar fields of individual flow components using fluid dynamics operators. These methods are gradient-based and sensitive to data uncertainties, and only focus on individual flow components of the overall composite flow, therefore potentially underestimating the severity of overall flow changes associated with cardiovascular diseases. To address these limitations, in MICCAI 2021, we introduced a novel comprehensive stochastic 4D Flow vector-field signature technique that works directly on the entire spatiotemporal velocity vector field. This technique uses efficient stochastic gradient-free interrogation of multi-million flow vector-pairs per patient to derive the patient's unique flow profile of the complex composite flow alterations and in real-time processing. The signature technique's probabilistic gradient-free formulation should allow for highly robust quantification despite inherent errors in 4D flow MRI acquisitions. Here, we extend the application of the 4D flow vector-field signature technique to the left atrium to analyze complex composite flow changes in patients with atrial fibrillation. In 128 subjects, we performed extensive sensitivity testing and determined that the vector-field signature technique is highly robust to typical sources of data uncertainties in 4D flow MRI: degradation in spatiotemporal resolution, added gaussian noise, and segmentation errors. We demonstrate the excellent generalizability of the stochastic convergence from the aorta to the left atrium and between different 4D Flow MRI acquisition protocols. We compare the robustness of our technique to existing advanced flow quantification metrics of kinetic energy, vorticity, and energy loss demonstrating a superior performance of up-to 14-fold. Our results show the potential diagnostic and clinical utility of our signature technique in identifying distinctly altered composite flow signatures in atrial fibrillation patients independent of existing flow metrics.

The diagnostic performance of micro-RNA and metabolites in lung cancer: A meta-analysis.

The diagnosis of lung cancer is based on the microscopic exam of tissue or liquid. During the recent decade, many biomarkers have been pointed to have a potential diagnostic role. These biomarkers may be assessed in blood, pleural effusion or sputum and they could avoid biopsies or other risky procedures. The authors aimed to assess the diagnostic performances of biomarkers focusing on micro-RNA and metabolites. This meta-analysis was conducted under the PRISMA guidelines during a nine-year-period (2013-2022). the Meta-Disc software 5.4 (free version) was used. Q test and I2 statistics were carried out to explore the heterogeneity among studies. Meta-regression was performed in case of significant heterogeneity. Publication bias was assessed using the funnel plot test and the Egger's test (free version JASP). According to our inclusion criteria, 165 studies from 79 articles were included. The pooled SEN, SPE and dOR accounted, respectively, for 0.76, 0.79 and 13.927. The AUC was estimated to 0.859 suggesting a good diagnostic accuracy. The heterogeneity in the pooled SEN and SPE was statistically significant. The meta-regression analysis focusing on the technique used, the sample, the number of biomarkers, the biomarker subtype, the tumor stage and the ethnicity revealed the biomarker number (p = 0.009) and the tumor stage (p = 0.0241) as potential sources of heterogeneity. Even if this meta-analysis highlighted the potential diagnostic utility of biomarkers, more prospective studies should be performed, especially to assess the biomarkers' diagnostic potential in early-stage lung cancers.

S100 Protein Expression in Primary and Metastatic Neuroendocrine Neoplasms: A Specific Marker of Pancreatic Origin.

Neuroendocrine neoplasms can arise in a wide variety of anatomic sites including the gastrointestinal tract, pancreas, and lung, among others. Here, we report on the expression of S100 protein in a tissue microarray composed of 919 distinct primary and metastatic neuroendocrine neoplasms from 548 patients. S100 protein is a commonly used marker in many laboratories for the identification of neural and melanocytic neoplasms and occasionally used in the workup for neuroendocrine neoplasms when the diagnosis of paraganglioma is being considered. We show that strong S100 protein expression is highly specific to well-differentiated neuroendocrine tumors of pancreatic origin. This finding suggests potential diagnostic utility of this marker in cases of tumors of unknown origin, and emphasizes that S100 protein expression should not be an unexpected finding in neuroendocrine tumors of pancreatic origin.

The role of noncoding RNA and its diagnostic potential in intrahepatic cholestasis of pregnancy: a research update.

Intrahepatic cholestasis of pregnancy (ICP) is a common liver disorder that generally occurs during the second or third trimester of pregnancy. It rarely causes any harm to the mother; however, it can result in short- and long-term complications in the offspring. Therefore, it is crucial to diagnose and treat this condition to avoid poor pregnancy outcomes. The identification of novel markers with potential diagnostic, prognostic, and therapeutic utility in ICP has gained attention. Noncoding RNAs (ncRNAs), including microRNA, long noncoding RNA, and circular RNA, are a type of transcripts that are not translated into proteins. They possess vital biological functions, including transcriptional and translational regulation and DNA, RNA, and protein interactions. The pathogenesis of ICP is related to the aberrant expression of several circulating or placenta-related ncRNAs. In this review, we summarized all recent findings on ncRNAs and ICP and outlined the concepts that form the basis for the early diagnosis and targeted treatment of ICP.

Altered expression of the L-arginine/nitric oxide pathway in ovarian cancer: metabolic biomarkers and biological implications

MotivationOvarian cancer (OC) is a highly lethal gynecological malignancy. Extensive research has shown that OC cells undergo significant metabolic alterations during tumorigenesis. In this study, we aim to leverage these metabolic changes as potential biomarkers for assessing ovarian cancer.MethodsA functional module-based approach was utilized to identify key gene expression pathways that distinguish different stages of ovarian cancer (OC) within a tissue biopsy cohort. This cohort consisted of control samples (n = 79), stage I/II samples (n = 280), and stage III/IV samples (n = 1016). To further explore these altered molecular pathways, minimal spanning tree (MST) analysis was applied, leading to the formulation of metabolic biomarker hypotheses for OC liquid biopsy. To validate, a multiple reaction monitoring (MRM) based quantitative LCMS/MS method was developed. This method allowed for the precise quantification of targeted metabolite biomarkers using an OC blood cohort comprising control samples (n = 464), benign samples (n = 3), and OC samples (n = 13).ResultsEleven functional modules were identified as significant differentiators (false discovery rate, FDR < 0.05) between normal and early-stage, or early-stage and late-stage ovarian cancer (OC) tumor tissues. MST analysis revealed that the metabolic L-arginine/nitric oxide (L-ARG/NO) pathway was reprogrammed, and the modules related to "DNA replication" and "DNA repair and recombination" served as anchor modules connecting the other nine modules. Based on this analysis, symmetric dimethylarginine (SDMA) and arginine were proposed as potential liquid biopsy biomarkers for OC assessment. Our quantitative LCMS/MS analysis on our OC blood cohort provided direct evidence supporting the use of the SDMA-to-arginine ratio as a liquid biopsy panel to distinguish between normal and OC samples, with an area under the ROC curve (AUC) of 98.3%.ConclusionOur comprehensive analysis of tissue genomics and blood quantitative LC/MSMS metabolic data shed light on the metabolic reprogramming underlying OC pathophysiology. These findings offer new insights into the potential diagnostic utility of the SDMA-to-arginine ratio for OC assessment. Further validation studies using adequately powered OC cohorts are warranted to fully establish the clinical effectiveness of this diagnostic test.

Unraveling the Role of EV-Derived miR-150-5p in Prostate Cancer Metastasis and Its Association with High-Grade Gleason Scores: Implications for Diagnosis.

Metastasis remains the leading cause of mortality in prostate cancer patients. The presence of tumor cells in lymph nodes is an established prognostic indicator for several cancer types, such as melanoma, breast, oral, pancreatic, and cervical cancers. Emerging evidence highlights the role of microRNAs enclosed within extracellular vesicles as facilitators of molecular communication between tumors and metastatic sites in the lymph nodes. This study aims to investigate the potential diagnostic utility of EV-derived microRNAs in liquid biopsies for prostate cancer. By employing microarrays on paraffin-embedded samples, we characterized the microRNA expression profiles in metastatic lymph nodes, non-metastatic lymph nodes, and primary tumor tissues of prostate cancer. Differential expression of microRNAs was observed in metastatic lymph nodes compared to prostate tumors and non-metastatic lymph node tissues. Three microRNAs (miR-140-3p, miR-150-5p, and miR-23b-3p) were identified as differentially expressed between tissue and plasma samples. Furthermore, we evaluated the expression of these microRNAs in exosomes derived from prostate cancer cells and plasma samples. Intriguingly, high Gleason score samples exhibited the lowest expression of miR-150-5p compared to control samples. Pathway analysis suggested a potential regulatory role for miR-150-5p in the Wnt pathway and bone metastasis. Our findings suggest EV-derived miR-150-5p as a promising diagnostic marker for identifying patients with high-grade Gleason scores and detecting metastasis at an early stage.

Potential Diagnostic Utility of microRNAs in Gastrointestinal Cancers.

Early detection of gastrointestinal cancers is beneficial for patient survival and prognosis. MiRNAs have been shown to be potential cancer biomarkers that can be used to diagnose cancers. MiRNAs are single-stranded, small non-coding RNAs that are involved in the post-transcriptional regulation of the expression of different oncogenes. Cancer tissues contain miRNAs that play a special role in the etiology of cancer development or limiting cancer suppression. Dysregulation of miRNAs occurs in a variety of malignancies, including gastrointestinal cancers. MiRNAs are stable and protected against degradation by RNase, which enables their detection in tissues and biological fluids. The results of many studies suggest that miRNAs have a relatively higher diagnostic efficiency in distinguishing cancer patients from healthy people. The researchers have identified many miRNA signature in the blood for the detection of gastrointestinal cancers. This review focuses on the role and potential utility of miRNAs in the early detection, prognosis and evaluation of the treatment effectiveness of gastrointestinal cancers.

The performance of Carbohydrate Antigen 125-Thomsen-nouveau and anti-Müllerian hormone combined with CA125, Human epididymis protein 4 and Risk of Malignancy Algorithm in diagnosis for patients with Epithelial ovarian cancer

ObjectivesWe examined the blood concentrations of Carbohydrate Antigen 125-Thomsen-nouveau (CA125-Tn) and anti-Müllerian hormone (AMH) in epithelial ovarian cancer (EOC) patients to evaluate their potential diagnostic utility together with CA125, human epididymis protein 4 (HE4) and Risk of Malignancy Algorithm (ROMA). Design & methods50 healthy subjects, 45 EOC patients, 22 patients with borderline ovarian tumors (BOT), 21 patients with benign ovarian tumor (BET) and 45 patients with chocolate cyst of ovary (CCO) were studied. Blood levels of CA125, HE4, CA125-Tn and AMH were measured, and the ROMA value was calculated. We compared the differences in the levels of these biomarkers among groups. Additionally, a total of 10 testing strategies were established for comparison to maximize the diagnostic value. ResultsThe levels of CA125, HE4, CA125-Tn and ROMA value were significantly higher in EOC group compared with either the disease control (DC) group (BOT group, BET group and CCO group) or healthy control (HC) group (p < 0.001). In addition, they had better discriminatory performance with an area under the receiver operator characteristic curve (AUC) 0.93; 0.93; 0.93; 0.85, respectively (p < 0.001) compared with the AUC value of AMH 0.67 (p < 0.001). Among all 10 testing strategies, both single-positive of ROMA and double-positive of any 2 markers showed better Youden index (0.82, 0.79, respectively) and kappa value (κ) (0.82, 0.81, respectively). ConclusionsCA125-Tn and AMH can be treated as useful biomarkers of EOC when combined with CA125, HE4 and ROMA, because when any two biomarkers of them are positive, the value of EOC diagnosis is maximized.

Emerging Concepts in Precision Medicine in Axial Spondyloarthritis.

Axial spondyloarthritis (AxSpA) is among the rheumatology's most heritable complex diseases, yet precision medicine at clinics still needs to be explored. We reviewed the emerging concepts and recent developments in polygenic risk scores, Mendelian randomization, pharmacogenomics, single-cell sequencing, and spatial transcriptomics. Polygenic risk score has resulted in encouraging results with potential diagnostic utility as it appears to outperform current diagnostic tools. Its performance and generalizability vary with ethnicity. Mendelian randomization has elucidated multiple causal associations, particularly between inflammatory bowel disease and AxSpA. Single-cell transcriptomics (particularly scRNA-seq and scATAC-seq) has identified numerous cell types, including synovial and blood immunological cells, to understand the contribution of both innate and adaptative immunity in AxSpA. Current molecular tools provide an exciting opportunity to advance precision medicine for AxSpA patients. However, extensive research and implementation strategies are still required before they can have an impact in the clinic.

CELA3B immunostaining is a highly specific marker for acinar cell carcinoma of the pancreas.

Chymotrypsin-like elastase family member 3B (CELA3B, elastase-3B) is a pancreatic enzyme with digestive function in the intestine. Since RNA analyses of normal tissues suggest that CELA3B expression is limited to the pancreas, the potential diagnostic utility of CELA3B immunohistochemistry for the distinction of pancreatic from extrapancreatic cancers and in the distinction of acinar cell carcinoma from ductal adenocarcinoma was assessed. CELA3B expression was successfully analyzed in 13,223 tumor samples from 132 different tumor types and subtypes as well as 8 samples each of 76 different normal tissue types by immunohistochemistry in a tissue microarray format (TMA). In normal tissues, CELA3B immunostaining was only seen in acinar cells and in a fraction of ductal cells of the pancreas as well as on some apical membranes of surface epithelial cells of the intestine. Among tumors, CELA3B immunostaining was seen in 12 of 16 (75%) acinar cell carcinoma of the pancreas including 6 cases with strong staining (37.5%) as well as in 5 of 13,207 other tumors (0.04%). These included 1.2% of 91 adenoid cystic carcinomas, 1.2% of 246 mucoepidermoid carcinomas and 0.8% of 127 acinic cell carcinomas of salivary glands. Our data show a good sensitivity (75%) and a high specificity (99.9%) of CELA3B immunohistochemistry for diagnosing acinar cell carcinoma of the pancreas.

Dissection of schistosome tissues under LC-MS compatible preservative conditions for quantitative proteomics.

Schistosomes are blood flukes with specialised tissues and organs, each one playing a pivotal role in perpetuating the parasite life cycle. Herein, we describe a detailed methodology for preserving the proteome of adult Schistosoma mansoni worms during manual dissection for enrichment of tissues associated with the parasite's alimentary tract. We provide step-by-step directions for specimen storage and dissection while in preservative solution, tissue homogenisation, protein extraction and digestion using a methodology fully compatible with downstream quantitative liquid chromatography-mass spectrometry analysis. Our methodology uses label-free and QconCAT-based absolute quantification for detection of S. mansoni oesophageal gland products proposed as vaccine candidates. Through stabilisation of the proteome and minimising sample degradation during dissection our approach has allowed us to access the hidden proteome of target tissues not readily available from total lysates because of their small volume. This protocol can be replicated or adapted to other Schistosoma species lacking quantitative proteomics characterisation of specialised tissues for discovery of proteins with potential diagnostic and therapeutic utility.

#5436 CHARACTERIZATION OF KIDNEY FUNCTIONAL DAMAGE ASSOCIATED TO CO-TREATMENT WITH IMMUNE CHECKPOINT INHIBITORS AND CISPLATIN

Abstract Background and Aims Advances in the knowledge of the immune response against tumors and the evasion mechanisms of these have led to develop new cancer therapies focused on improving the immune response and thus increasing patient survival. This is the case of immunotherapeutic treatment based on immune checkpoint inhibitors (ICI). However, treatment based on these antibodies is associated with autoimmune side effects, in most of the body organs, that limit their use. Although nephrotoxicity is rare, renal effects have been shown to worsen the prognosis of cancer patients. Recently, therapies based on the combination of immuno and chemotherapy have been approved. They have improved efficacy but have increased the risk of suffering nephrotoxic adverse effects. Our hypothesis is that kidney damage associated with ICIs could be subclinical and not evidenced by the parameters used in clinical practice (mainly plasma creatinine). The aim of this work was to characterize functional kidney damage associated with ICI (anti-CTLA-4) in combination with chemotherapy (cisplatin) in a murine model. Method C57BL/6 mice were treated with combined therapy of cisplatin (10 mg/kg, single dose) and anti-CTLA-4 (10 or 15 mg/kg/day, for 6 days) administrated by intraperitoneal injection. In addition, groups treated with drug monotherapies and a control group were included. Urine and blood samples were collected at baseline, on day 3 and on day 6 (sacrifice). Biomarkers of subclinical kidney damage were determined by ELISA in urine samples. There were albumin, neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule 1 (KIM-1). Blood samples were centrifuged to obtain plasma, in which creatinine and urea were measured using colorimetric techniques. Data were analyzed with the statistical software SPSS®. Results Plasma creatinine and urea were elevated in the combined therapy groups with respect to the monotherapies. However, these parameters were similar to the normal range, so they do not reflect obvious kidney injury. It was evidenced a statistically significant elevation of biomarkers of subclinical kidney damage in the combined therapy groups with respect to monotherapies and control group. Conclusion This study suggests that the combination of anti-CTLA-4 with cisplatin could cause a potenciation effect on subclinical kidney damage compared to monotherapies. This experimental model would make possible to study the nephrotoxicity mechanisms of ICI. Moreover, it could have a potential diagnostic utility through the identification of biomarkers of subclinical kidney damage, that would avoid biopsy.

Glycoprofiling of early non-small cell lung cancer using lectin microarray technology

Abstract Objectives Non-small cell lung cancer (NSCLC) is one of the most common malignancies in the world with a high incidence and it lacks effective biomarkers for early-stage detection. In this investigation, we aimed to investigate the alterations in plasma glycans related to NSCLC and assess the possibility of plasma glycopatterns as potential biomarkers for the diagnosis of NSCLC. Methods First, plasma samples from 16 patients with early-stage lung adenocarcinoma (LUAD), 16 patients with early-stage Lung squamous cell carcinoma (LUSC), and 16 healthy volunteers, were selected for inclusion in this study to probe the difference in plasma glycopatterns using lectin microarrays. Then, the diagnostic effectiveness of the candidate lectins was evaluated using ROC. Results In contrast to the NL group, seven candidate lectins offered potential diagnostic utility in the NSCLC (LUAD and LUSC) group. F17AG was significantly altered in LUSC with an AUC of 0.818 (adj.P.Val&lt;0.05) compared to NL samples. There were 20 differentially expressed lectins in the LUAD group compared to the NL group. Based on the AUC values (AUC&gt;0.800) and the normalized fluorescence intensities of the lectins, we selected eight lectins, GAL2, PTL-1, GNA, SSA, LENTIL, CA, PHA-E, and MAA to perform logistic regression analysis, and found that the combination of these eight candidate lectins had high diagnostic potential. Conclusions The results of this study should help to distinguish between NSCLC and NL based on changes in plasma glycopatterns, which have a great deal of potential to be biomarkers for diagnosing NSCLC.

Large retrospective validation study of metabolomic biomarkers for resectable lung cancer detection and risk assessment.

3071 Background: Cancer can be regarded as a metabolic disease and many studies published aimed at identifying robust metabolites for lung cancer diagnosis using plasma samples. Regardless of the lung cancer staging, most of these studies were performed on relatively small sample sizes. The purpose of this study is to validate whether a panel of metabolomic biomarkers would improve risk assessment for lung cancer detection in 800 plasma samples from patients that underwent lung cancer resection, and to understand the potential role and intersection between lung cancer and other lung diseases. Methods: A blinded case-control study was performed using plasma samples from 586 patients with biopsy-confirmed lung cancer compared to 214 controls from the same institutional biorepository to evaluate the performance of a 5+ metabolites biomarker panel for lung cancer detection. The control group consists of 90 healthy individuals and 124 with other non-neoplastic lung diseases including asthma, COPD, bronchiectasis and COVID. The lung cancer subgroups include early-stage (Stage I &amp;II) adenocarcinoma and squamous cells carcinoma, advanced stages (Stage III &amp; IV) NSCLC and neuroendocrine tumors. In this study, plasma metabolic profiles were performed using different liquid chromatography methods coupled with a targeted and quantitative mass spectrometry approach. Metabolite concentrations, clinical data, and smoking history were used to develop logistic regression models to identify lung cancer at different stages using significant biomarkers. The area under the receiver operator characteristic curves (AUC), sensitivities and specificities at selected cut off points were calculated for each cancer stage. Results: Univariate and multivariate statistical analyses confirmed the performance of our 5+ metabolites panel (β-HBA, PC aa C38:0, PC ae C40:6, citric acid, tryptophan, and etc.) as being significantly different between controls and lung cancer cases. Linear regression model using metabolites alone yielded an AUC of 0.89 for lung cancer at all stages. When adding smoking status, the models achieved an AUC of 0.91 with sensitivity of 91% and specificity over 78% using a risk prediction threshold of 0.657. Conclusions: The blood-based metabolites panel validated on our current retrospective study exhibited robust performance for resectable lung cancer detection and risk assessment. This metabolomic panel assay demonstrates potential diagnostic applications and clinical utility for patient selection that require further follow-up and confirmation using LDCT or other lung imaging modalities. The inclusion of other lung diseases in the control group is more representative of a real clinical setting suggesting that this blood-based assay could be offered to the medical community. Further studies may also confirm its utility in the context of a lung cancer screening program.

Evaluation of the Potential Diagnostic Utility of the Determination of Selected Immunological and Molecular Parameters in Patients with Ovarian Cancer.

Ovarian cancer is one of the most serious challenges in modern gynaecological oncology. Due to its non-specific symptoms and the lack of an effective screening procedure to detect the disease at an early stage, ovarian cancer is still marked by a high mortality rate among women. For this reason, a great deal of research is being carried out to find new markers that can be used in the detection of ovarian cancer to improve early diagnosis and survival rates of women with ovarian cancer. Our study focuses on presenting the currently used diagnostic markers and the latest selected immunological and molecular parameters being currently investigated for their potential use in the development of new diagnostic and therapeutic strategies.

Abstract 5555: Altered p53/p16 expression is linked to urothelial carcinoma progression but is unrelated to prognosis in muscle-invasive tumors

Abstract Background: Most inactivating p53 mutations result in a nuclear accumulation of the defective p53 protein. However, p53 alterations that result in a complete lack of cellular p53 and complete absence of p53 immunostaining do also occur. As p16 is upregulated in p53 inactivated cells, p16 immunohistochemistry may be a surrogate marker for p53 inactivation. Design: In this study, we investigated p53 and p16 immunostaining on more than 2,500 urothelial bladder carcinomas in a tissue microarray format to better understand their impact in relation to clinicopathological parameters of disease progression and patient outcome. Results: p16 immunostaining was not observed in normal urothelium but occurred in 1,576 (63.5%) of cancers including 755 (30.4%) with a strong staining. The fraction of p16 positive cases increased markedly from pTaG2 low grade (9.6%) to pTaG3 high grade tumors (46.5% strongly positive, p&amp;lt;0.0001 for pTaG2 low vs. pTaG3) but continuously decreased from pTaG3 to pT2 (41.3% strongly positive), pT3 (36.5%) and pT4 (33.3%; p=0.0030). Within pT2-4 carcinomas, p16 positivity was also linked to high grade (p=0.0005) but unrelated to overall survival. p53 staining has been recorded as negative in 203 (8.4%), very weak in 373 (15.4%), weak in 1,341 (55.3%), strong in 115 (4.7%), and very strong in 393 (16.2%) cancers. The fraction of tumors with negative (potentially p53 null phenotype), strong, and very strong p53 positivity increased markedly from pTaG2 low grade to pTaG3 high grade tumors (p&amp;lt;0.0001) and from pTaG3 to muscle-invasive pT2-4 cancers (p=0.0007). p53 staining pattern was unrelated to histopathological parameters of malignancy or patient prognosis within pT2-4 carcinomas, however. There was a significant overall association between p53 and p16 expression but strong p16 expression predominated in tumors with very strong, strong, and negative p53 staining. Subset analyses showed that the combination of p53 negative/p16 strongly positive cancers was particularly linked to features of tumor aggressiveness. Conclusion: Our data show that altered function of p53 and p16 immunostaining increases during grade and stage progression although these alterations lack prognostic significance in pT2-4 carcinomas. That high level p16 expression is limited to neoplastic urothelium and that the p53 null phenotype is largely limited to grade 3 and invasive urothelial carcinomas are features with potential diagnostic utility. Citation Format: Simon Schallenberg, Henning Plage, Sebastian Hofbauer, Kira Kornienko, Sarah Weinberger, Paul G. Bruch, Florian Roßner, Sefer Elezkurtaj, Martina Kluth, Maximilian Lennartz, Tim Mandelkow, Elena Bady, Niclas C. Blessin, Andreas H. Marx, Henrik Samtleben, Margit Fisch, Michael Rink, Marcin Slojewski, Krystian Kaczmarek, Thorsten Ecke, Steffen Hallmann, Stefan Koch, Nico Adamini, Sarah Minner, Ronald Simon, Guido Sauter, David Horst, Tobias Klatte, Thorsten Schlomm, Henrik Zecha. Altered p53/p16 expression is linked to urothelial carcinoma progression but is unrelated to prognosis in muscle-invasive tumors. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5555.

Pathogenic role of Twist-1 protein in hydatidiform molar pregnancies and investigation of its potential diagnostic utility in complete moles

BackgroundComplete and partial moles (PM) are the most common gestational trophoblastic diseases. Due to some overlapping morphological findings, ancillary studies may be necessary.MethodsIn this cross-sectional study, 47 cases of complete mole (CM) and 40 cases of PM were randomly selected based on histopathological criteria. Only those cases that were agreed upon by two expert gynecological pathologists and confirmed by the P57 IHC study were included. The expression level of the Twist-1 marker in villi stromal cells, as well as syncytiotrophoblasts, was evaluated quantitatively (percentage of positive cells), qualitatively (staining intensity) and as a total comprehensive score.ResultsExpression of Twist-1 is higher and more intense in villous stromal cells of CMs (p < 0.001). Moderate to strong staining intensity in more than 50% of villous stromal cells, can differentiate CM and PM with 89.5% sensitivity and 75% specificity. In syncytiotrophoblasts of CM, Twist-1 expression was significantly lower than PM (p < 0.001). Negative or weak staining intensity in less than 10% of syncytiotrophoblasts, can distinguish CM and PM with 82.9% sensitivity and 60% specificity.ConclusionA higher expression of Twist-1 in villous stromal cells of hydatidiform moles is a sensitive and specific marker for the diagnosis of CMs. An elevated expression of this marker in villous stromal cells suggests another pathogenic mechanism for more aggressiveness of CMs in addition to the characteristics of trophoblast cells. The opposite result was obtained in the expression of Twist-1 in the syncytiotrophoblasts, compatible with defects in the process of formation of these supportive cells in CMs.

A Comparative Analysis of Treatment-Related Changes in the Diagnostic Biomarker Active Metalloproteinase-8 Levels in Patients with Periodontitis.

Previous studies have revealed the potential diagnostic utility of aMMP-8, an active form of MMP-8, in periodontal and peri-implant diseases. While non-invasive point-of-care (PoC) chairside aMMP-8 tests have shown promise in this regard, there is a dearth of literature on the evaluation of treatment response using these tests. The present study aimed to investigate treatment-related changes in aMMP-8 levels in individuals with Stage III/IV-Grade C periodontitis compared to a healthy control group, using a quantitative chairside PoC aMMP-8 test, and to determine its correlation with clinical parameters. The study included 27 adult patients (13 smoker, 14 non-smoker) with stage III/IV-grade C periodontitis and 25 healthy adult subjects. Clinical periodontal measurements, real-time PoC aMMP-8, IFMA aMMP-8, and Western immunoblot analyses were performed before and 1 month after anti-infective scaling and root planing periodontal treatment. Time 0 measurements were taken from the healthy control group to test the consistency of the diagnostic test. Both PoC aMMP-8 and IFMA aMMP-8 tests showed a statistically significant decrease in aMMP-8 levels and improvement in periodontal clinical parameters following treatment (p < 0.05). The PoC aMMP-8 test had high diagnostic sensitivity (85.2%) and specificity (100.0%) for periodontitis and was not affected by smoking (p > 0.05). Treatment also reduced MMP-8 immunoreactivity and activation as demonstrated by Western immunoblot analysis. The PoC aMMP-8 test shows promise as a useful tool for the real-time diagnosis and monitoring of periodontal therapy.