Biomarkers For Detection Of Cancer Research Articles

Electrochemical Detection of a Breast Cancer Biomarker with an Amine-Functionalized Nanocomposite Pt-Fe3O4-MWCNTs-NH2 as a Signal-Amplifying Label.

We report an ultrasensitive sandwich-type electrochemical immunosensor to detect the breast cancer biomarker CA 15-3. Amine-functionalized composite of reduced graphene oxide and Fe3O4 nanoparticles (MRGO-NH2) was used as an electrochemical sensing platform material to modify the electrodes. The nanocomposite comprising Pt and Fe3O4 nanoparticles (NPs) anchored on multiwalled carbon nanotubes (Pt-Fe3O4-MWCNTs-NH2) was utilized as a pseudoenzymatic signal-amplifying label. Compared to reduced graphene oxide, the composite MRGO-NH2 platform material demonstrated a higher electrochemical signal. In the Pt-Fe3O4-MWCNTs-NH2 label, multiwalled carbon nanotubes provided the substratum to anchor abundant catalytic Pt and Fe3O4 NPs. The nanocomposites were thoroughly characterized using transmission electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, and X-ray photoelectron spectroscopy. An electroanalytical study and prevalidation of the immunosensor was carried out. The immunosensor exhibited exceptional capabilities in detecting CA 15-3, offering a wider linear range of 0.0005-100 U mL-1 and a lower detection limit of 0.00008 U mL-1. Moreover, the designed immunosensor showed good specificity, reproducibility, and acceptable stability. The sensor was successfully applied to analyze samples from breast cancer patients, yielding reliable results.

Cycling back to folate metabolism in cancer.

Metabolic changes contribute to cancer initiation and progression through effects on cancer cells, the tumor microenvironment and whole-body metabolism. Alterations in serine metabolism and the control of one-carbon cycles have emerged as critical for the development of many tumor types. In this Review, we focus on the mitochondrial folate cycle. We discuss recent evidence that, in addition to supporting nucleotide synthesis, mitochondrial folate metabolism also contributes to metastasis through support of antioxidant defense, mitochondrial protein synthesis and the overflow of excess formate. These observations offer potential therapeutic opportunities, including the modulation of formate metabolism through dietary interventions and the use of circulating folate cycle metabolites as biomarkers for cancer detection.

Abstract PO5-07-08: A Serum Biomarker for the Early Detection of Breast cancer

Abstract Background: After decades of use, clinicians and medical societies are still debating on the clinical utility of mammography, the standard of care for breast cancer (BC) screening, because of its limitations such as subjective visual interpretation, high rate of false-positive and false-negative (in dense breast) and concerns over radiation exposure. There are 55 million women, and about 45% of them considered dense breast, in need of an accurate, easy to use test for the screening of BC regardless of breast tissue complexities to differentiate between BC and benign findings such as calcification and fibrosis. To this date there is no blood protein biomarker (BM) in clinical use for the early detection of BC. We have developed a novel BC specific protein biomarker -based blood test to detect patients with early-stage BC and established herein the diagnostic performance of our BM in discriminating BC patients from healthy individuals and benign breast cases. Methods: A total of 433 serum samples (318 primary BC, 32 benign breast and 83 normal breast) were obtained from the Cooperative Human Tissue Network and from the IRCCS San Raffaele Pisana Research Center (blind samples). 247 BC samples were provided as blind samples and were used to validate the screening utility. Serum samples were tested with our BC specific BM by sandwich ELISA. Receiver operating characteristic (ROC) curves were calculated to evaluate the diagnostic performance, by determining sensitivity (SE), specificity (SP), area under the curve (AUC), and confidence interval (CI). Statistical analysis, ROC curves and scatter plots were performed with GraphPad Prism 7.0. Results: BM elevation in early cancer cases (stage I-II, Ductal Carcinoma) was found to be statistically significant (p< 0.0001) as compared to healthy and benign cases. Our test discriminated early BC patients from non-cancer cases with 87.3% SE at 85.2% SP and was further validated in a blind set including 247 BC (Ductal & Lobular Carcinoma, stage I-III), 30 benign and 83 normal breast cases (88%SE, 86%SP). The sensitivity for early-stage (I-II) and late stage (III) BC was 86.8% and 94.3%, respectively. While lobular carcinoma is particularly not well detected by mammography with a sensitivity as low as 57% and much lower for women with dense breast, it was detected with high sensitivity (90%) in our test. 80% of fibrocystic changes (FCC), the most common form of benign breast disease in women over 50 years of age were negative with our test. Although it is a benign condition, FCC may be misdiagnosed as a malignant lesion by physical examination and imaging. Conclusion: Our results support the utility of our blood-based test for the early detection of breast cancer which presents improved features over mammography such as higher lobular cancer sensitivity, lower detection rate of common form of benign condition often misdiagnosed by imaging, absence of radiation and expected high compliance. Further testing is needed to confirm accuracy of our test in the dense breast population. Citation Format: Christine Chavany, Jeffrey Dea, Monica Guevara, Rafael Hernandez-Gonzalez, Patrizia Ferroni, Fiorella Guadagni, Rosaura Valle, Moncef Jendoubi. A Serum Biomarker for the Early Detection of Breast cancer [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-07-08.

Investigating alterations in amino acid levels and immune cell population as potential biomarkers for non-invasive early detection of breast cancer: Insights from a study in north India

Investigating alterations in amino acid levels and immune cell population as potential biomarkers for non-invasive early detection of breast cancer: Insights from a study in north India

Circulating MiR-1185¸ MiR-3183, and MiR-584 Levels in Plasma as Diagnostic Non-invasive Biomarkers for Early Detection of Breast Cancer

Introduction: MicroRNAs (miRNAs) are non-coding RNA molecules with short sequences that function as main post-transcriptional gene regulators of different biological pathways via negative regulation of gene expression, thereby leading to either mRNA degradation or translational blockade. Dysregulated expression of these miRNAs has been related etiologically to many human diseases, including breast cancer. Various cellular processes of breast cancer progression, including cell proliferation, apoptosis, metastasis, recurrence and chemodrug resistance, are modulated by oncogenic miRNA (oncomiR). Objective: The objective of this investigation was to study the expression level and potential diagnostic/ prognostic roles of circulating microRNAs (miR-3183, miR-1185, and miR-584) as novel breast cancer biomarkers. Method: The current study was conducted on 99 breast cancer (BC) female patients, aged between 20-63 years old, as the case group and 50 age-matched healthy females as control (HC). After microRNA extraction from the serum samples, real-time PCR was carried out for relative expression quantification of miR-1185, miR-3183a, and miR-584. The ROC curve analysis was performed to investigate the diagnostic value of miRNAs. Result: It was demonstrated that miRNA-1185, miRNA-584, and miRNA-3183 were significantly up-regulated (p-values <0.0001) in female BC cases compared to the control group. Besides, based on the ROC analysis for BC versus HC, it was revealed that the AUC for miRNA-584 was 0.844 (95% confidence interval (CI) and could be proposed as a diagnostic biomarker for breast cancer screening and follow-up. Conclusion: MiRNAs expression profiling using blood-based samples demonstrated their upregulation in the serum and plasma and revealed the concept that circulating miRNAs have high potential as novel noninvasive biomarkers for cancer diagnosis and screening.

Role of MicroRNA in Hypoxic Tumours and their Potential as Biomarkers for Early Detection of Cancer.

Hypoxia is a pathophysiological condition characterized by oxygen deficiency in tissues, which negatively affects normal biological functions. It is a typical microenvironment character of almost all solid tumours. Noncoding RNA are small functional RNA molecules that regulate gene expression at chromatin and posttranscriptional levels. Micro-RNAs (miRNAs) are a type of noncoding RNA and are ~12-22 nucleotides long that are crucial in regulating gene expression by partnering with the mRNAs of protein-coding genes. It is widely reported that miRs play an important role in various key processes and pathways during tumour formation, as well as advancement in hypoxic tumors by influencing the HIF pathway. The role of miRNAs in hypoxic tumours, namely in pancreatic, kidney, breast, lung and colorectal, are described. These miRNAs have immense potential as diagnostic and prognostic biomarkers for early cancer detection.

Recent Trends and Innovations in Bead-Based Biosensors for Cancer Detection.

Demand is strong for sensitive, reliable, and cost-effective diagnostic tools for cancer detection. Accordingly, bead-based biosensors have emerged in recent years as promising diagnostic platforms based on wide-ranging cancer biomarkers owing to the versatility, high sensitivity, and flexibility to perform the multiplexing of beads. This comprehensive review highlights recent trends and innovations in the development of bead-based biosensors for cancer-biomarker detection. We introduce various types of bead-based biosensors such as optical, electrochemical, and magnetic biosensors, along with their respective advantages and limitations. Moreover, the review summarizes the latest advancements, including fabrication techniques, signal-amplification strategies, and integration with microfluidics and nanotechnology. Additionally, the challenges and future perspectives in the field of bead-based biosensors for cancer-biomarker detection are discussed. Understanding these innovations in bead-based biosensors can greatly contribute to improvements in cancer diagnostics, thereby facilitating early detection and personalized treatments.

Circulating tumor DNA: from discovery to clinical application in breast cancer.

Breast cancer (BC) stands out as the cancer with the highest incidence of morbidity and mortality among women worldwide, and its incidence rate is currently trending upwards. Improving the efficiency of breast cancer diagnosis and treatment is crucial, as it can effectively reduce the disease burden. Circulating tumor DNA (ctDNA) originates from the release of tumor cells and plays a pivotal role in the occurrence, development, and metastasis of breast cancer. In recent years, the widespread application of high-throughput analytical technology has made ctDNA a promising biomarker for early cancer detection, monitoring minimal residual disease, early recurrence monitoring, and predicting treatment outcomes. ctDNA-based approaches can effectively compensate for the shortcomings of traditional screening and monitoring methods, which fail to provide real-time information and prospective guidance for breast cancer diagnosis and treatment. This review summarizes the applications of ctDNA in various aspects of breast cancer, including screening, diagnosis, prognosis, treatment, and follow-up. It highlights the current research status in this field and emphasizes the potential for future large-scale clinical applications of ctDNA-based approaches.

Stool protein mass spectrometry identifies biomarkers for the early detection of diffuse-type gastric cancer.

There is a high unmet need for early detection approaches for diffuse gastric cancer (DGC). We examined whether the stool proteome of mouse models of GC or individuals with hereditary diffuse GC (HDGC) have utility as biomarkers for early detection. Proteomic mass spectrometry of stool from a genetically engineered mouse model driven by oncogenic KrasG12D and loss of p53 and Cdh1 in gastric parietal cells (known as TCON mice) identified differentially abundant proteins compared to littermate controls. Immunoblot assays validated a panel of proteins including actinin alpha 4 (ACTN4), N-acylsphingosine amidohydrolase 2 (ASAH2), dipeptidyl peptidase 4 (DPP4), and valosin-containing protein (VCP) as enriched in TCON stool compared to littermate control stool. Immunofluorescence analysis of these proteins in TCON stomach sections revealed increased protein expression as compared to littermate controls. Proteomic mass spectrometry of stool obtained from HDGC patients with CDH1 mutations identified increased expression of ASAH2, DPP4, VCP, lactotransferrin (LTF), and tropomyosin-2 (TPM2) relative to stool from healthy sex and age-matched donors. Chemical inhibition of ASAH2 using C6-urea ceramide was toxic to GC cell lines and patient derived-GC organoids. This toxicity was reversed by adding downstream products of the S1P synthesis pathway, suggesting a dependency on ASAH2 activity in GC. An exploratory analysis of the HDGC stool microbiome identified features which correlated with patient tumors. Here we provide evidence supporting the potential of analyzing stool biomarkers for the early detection of DGC.

Cancer Methylation Biomarker Detection in an Automated, Portable, Multichannel Magnetofluidic Platform.

Early detection of cancer is critical to improving clinical outcomes, especially in territories with limited healthcare resources. DNA methylation biomarkers have shown promise in early cancer detection, but typical workflows require highly trained personnel and specialized equipment for manual and lengthy processing, limiting use in resource-constrained areas. As a potential solution, we introduce the Automated Cartridge-based Cancer Early Screening System (ACCESS), a compact, portable, multiplexed, automated platform that performs droplet magnetofluidic- and methylation-specific qPCR-based assays for the detection of DNA methylation cancer biomarkers. Development of ACCESS focuses on esophageal cancer, which is among the most prevalent cancers in low- and middle-income countries with extremely low survival rates. Upon implementing detection assays for two esophageal cancer methylation biomarkers within ACCESS, we demonstrated successful detection of both biomarkers from esophageal tumor tissue samples from eight esophageal cancer patients while showing specificity in paired normal esophageal tissue samples. These results illustrate ACCESS's potential as an amenable epigenetic diagnostic tool for resource-constrained areas toward early detection of esophageal cancer and potentially other malignancies.

The role of biomarkers in the early detection of pancreatic cancer.

Pancreatic surveillance can detect early-stage pancreatic cancer and achieve long-term survival, but currently involves annual endoscopic ultrasound and MRI/MRCP, and is recommended only for individuals who meet familial/genetic risk criteria. To improve upon current approaches to pancreatic cancer early detection and to expand access, more accurate, inexpensive, and safe biomarkers are needed, but finding them has remained elusive. Newer approaches to early detection, such as using gene tests to personalize biomarker interpretation, and the increasing application of artificial intelligence approaches to integrate complex biomarker data, offer promise that clinically useful biomarkers for early pancreatic cancer detection are on the horizon.

Electrospun EU/HPMC nanofibers decorated by ZIF-8 nanoparticle as the advanced electrochemical biosensor modifier for sensitive and selective detection of c-MET cancer biomarker in human plasma sample

This research presents a selective and sensitive electrochemical biosensor for the detection of the mesenchymal–epithelial transition factor (c-MET). The biosensing is based on a modification of the SPCE (screen-printed carbon electrode) with the electrospun nanofiber containing eudragit (EU), hydroxypropyl methylcellulose (HPMC), and Zeolite imidazolate frameworks (ZIF-8) nanoparticles. EU/HPMC/ZIF-8 nanofibers have presented a high capability of electron transfer, and more active surface area than bare SPCE due to synergistic effects between EU, HPMC, and ZIF-8. On the other hand, EU/HPMC nanofibers provided high porosity, flexible structures, high specific surface area, and good mechanical strength. The presence of ZIF-8 nanoparticles improved the immobilization of anti-c-MET on the modified SPCE and also resulted in increasing the conductivity. By c-MET incubation on the modified SPCE, c-MET was connected to anti-c-MET, and consequently the electrochemical signal of [Fe(CN)6]3-/4- as the anion redox probe was reduced. In order to investigate the structural and morphological characteristics and elemental composition of electrospun nanofibers, various characterization methods including FE-SEM, XRD, FTIR, and EDS were used. Under optimum conditions with a working potential range −0.3−0.6 V (vs. Ag/AgCl), linear range (LR), correlation coefficient (R2), sensitivity, and limit of detection (LOD) were acquired at 100 fg/mL-100 ng/mL, 0.9985, 53.28 μA/cm2.dec, and 1.28 fg/mL, respectively. Moreover, the mentioned biosensor was investigated in a human plasma sample to determine c-MET and showed ideal results including reproducibility, stability, and good selectivity against other proteins.

Development of a labeled-free and labeled electrochemical aptasensor for the detection of cancer antigen 125 by using magnetic g-C3N4/MoS2 nanocomposite

Efficient and timely detection of cancer biomarkers is pivotal for enhancing treatment outcomes and mitigating patient mortality. This study addresses the pressing need for a swift, accurate, and non-invasive method to identify cancer antigen 125 (CA125), a vital biomarker in ovarian cancer. Leveraging the growing prominence of nano-biosensors for their high selectivity and sensitivity, we present the development and characterization of an innovative electrochemical nano-biosensor. The sensor, featuring aptamer strands immobilized on a glassy carbon electrode modified with graphitic carbon nitrides, molybdenum disulfide, and magnetic nanoparticles (g-C3N4/MoS2/Fe3O4), demonstrates superior sensitivity and accuracy in CA125 detection. Utilizing methylene blue for electrochemical detection of labeled CA125 and ferrocyanide for label-free detection, our aptasensor achieves a low limit of detection (LOD) at 0.202 U.mL−1 and 0.215 U.mL−1, respectively, with a broad detection range from 2 to 10 U.mL−1. The modified electrode exhibits a pronounced affinity for CA125, demonstrating enhanced stability compared to other biomolecules. Crucially, the evaluation of both patient and normal serum samples underscores the aptasensor's remarkable performance. These findings not only establish a robust foundation for future research in ovarian cancer diagnosis but also highlight the potential clinical impact of our electrochemical nano-biosensor in advancing early cancer detection methodologies.

Fabrication of and characterization of directional antibody-conjugated gold nanourchin colloid and effect of laser polarization on SERS detection of breast cancer biomarker in serum

We describe the results of citrate-stabilized 100 nm gold nanourchins (GNU) functionalized and conjugated with monoclonal antibody (mAb) in directional orientation to investigate the interaction with breast cancer serum (BCS) overexpressed with CA 15–3 biomarker. UV-Vis, fluorescence, FT-NIR spectroscopy, and SERS with 637 nm Raman probe were employed for non-destructive analysis. UV-Vis spectroscopy exhibited two distinct peaks at 230 and 280 nm related to tyrosine (Tyr) and tryptophan (Trp) with the corresponding fluorescence peaks at 285 and 315 nm respectively. FT-NIR showed some new lines after the interaction such as CH2 combination in carbohydrates, and NH2 amino acids. Directional orientation indicated a higher SERS intensity than random due to higher parallel dipole moments of asymmetric molecules and the polarization sensitivity of GNU. The SERS signals increased significantly after the interaction and the intensity of the Raman lines varied nonlinearly with the laser power. The data were filtered using the filtfilt filter from scipy.signals, baseline corrected using the Improved Asymmetric Least Squares (isals) function from the pybaselines.Whittaker library, and normalized using the minmax_scale function from sklearn.preprocessing. The scikit-learn Principal Component Analysis (PCA) function was used to analyze the dataset by identifying the directions of maximum variability in the data.

Detecting androgen receptor (AR), AR variant 7 (AR-V7), prostate-specific membrane antigen (PSMA), and prostate-specific antigen (PSA) gene expression in CTCs and plasma exosome-derived cfRNA in patients with metastatic castration-resistant prostate cancer (mCRPC) by integrating the VTX-1 CTC isolation system with the QIAGEN AdnaTest

BackgroundTherapies for metastatic castration-resistant prostate cancer (mCRPC) include targeting the androgen receptor (AR) with androgen receptor inhibitors (ARIs) and prostate-specific membrane antigen (PSMA). Having the ability to detect AR, AR splice variant 7 (AR-V7), or PSMA in circulating tumor cells (CTCs) or circulating exosomal cell-free RNA (cfRNA) could be helpful to guide selection of the appropriate therapy for each individual patient. The Vortex Biosciences VTX-1 system is a label-free CTC isolation system that enables the detection of the expression of multiple genes in both CTCs and exosomal cfRNA from the same blood sample in patients with mCRPC. Detection of both AR-V7 and PSMA gene expression in both CTCs and cfRNA simultaneously has not yet been reported.MethodsTo characterize the combined VTX-1-AdnaDetect workflow, 22Rv1 cancer cells were spiked into blood from healthy donors and processed with the VTX-1 to mimic patient samples and assess performances (capture efficiency, purity, AR and AR-V7 expression). Then, we collected 19 blood samples from 16 patients with mCRPC and therapeutic resistance to androgen receptor inhibitors (ARIs). Plasma was separated and the plasma-depleted blood was processed further with the VTX-1 to collect CTCs. Both plasma exosomal cfRNA and CTCs were subsequently analyzed for AR, AR-V7, PSMA, and prostate-specific antigen (PSA) mRNA expression using the AdnaTest ProstateCancerPanel AR-V7 assay.ResultsAR-V7 expression could be detected in 22Rv1 cells spiked into blood from healthy volunteers as well as in CTCs and plasma-derived exosomal cfRNA from patients with mCRPC by processing blood with the VTX-1 CTC isolation system followed by the AdnaTest ProstateCancerPanel AR-V7 assay. 94.7% of patient blood samples (18/19) had detectable AR expression in either CTCs or exosomal cfRNA (16 in CTCs, 12 in cfRNA). 15.8% of the 19 patient blood samples (3/19) were found to have AR-V7-positive (AR-V7+) CTCs, one of which was also AR-V7+ in the exosomal cfRNA analysis. 42.1% of patient blood samples (8/19) were found to be PSMA positive (PSMA+): 26.3% (5/19) were PSMA+ in the CTC analysis and 31.6% (6/19) were PSMA+ in the exosomal cfRNA analysis. Of those 8 PSMA+ samples, 2 had detectable PSMA only in CTCs, and 3 had detectable PSMA only in exosomal cfRNA.ConclusionVTX-1 enables isolation of CTCs and plasma exosomes from a single blood draw and can be used for detecting AR-V7 and PSMA mRNA in both CTCs and cfRNA in patients with mCRPC and resistance to ARIs. This technology facilitates combining RNA measurements in CTCs and exosomal cfRNA for future studies to develop potentially clinically relevant cancer biomarker detection in blood.

Exploring research progress in studying serum exosomal miRNA-21 as a molecular diagnostic marker for breast cancer.

Breast cancer is one of the most prevalent malignancies affecting women globally and poses a significant public health challenge. Early clinical detection plays a pivotal role in providing optimal treatment opportunities and favorable prognoses, crucial for reducing breast cancer mortality and enhancing patients' quality of life. Therefore, the timely identification and diagnosis of breast cancer are imperative. Conventional tumor markers, such as carcinoembryonic antigen (CEA) and carbohydrate antigen 15-3 (CA15-3), serve as reliable methods for actively monitoring disease progression and have become a routine auxiliary diagnostic approach in clinical settings. However, these biomarkers exhibit limitations in sensitivity and specificity, particularly in the early screening and diagnosis of tumors, often yielding results inconsistent with clinical manifestations. In recent years, research has increasingly focused on exosomes released by tumor cells as potential new biomarkers for early stage breast cancer screening. Exosomes carry various components, including tumor-derived proteins, nucleic acids, and lipids. This paper delves into the specific utilization of serum exosomal microRNA-21 (miR-21) as a biomarker for early detection and diagnosis of breast cancer, evaluating its efficacy within this framework.

Genome-wide 5-hydroxymethylcytosines in circulating cell-free DNA as noninvasive diagnostic markers for gastric cancer.

5-Hydroxymethylcytosine-enriched gene profiles and regions show tissue-specific and tumor specific. There is a potential value to explore cell-free DNA 5-hydroxymethylcytosine feature biomarkers for early gastric cancer detection. A matched case‒control study design with 50 gastric cancer patients and 50 controls was performed to sequence the different 5-hydroxymethylcytosine modification features of cell free DNA. Significantly differential 5-hydroxymethylcytosine modification genes were identified to construct a gastric cancer diagnostic model. Data set from GEO was used as an external testing set to test the robustness of the diagnostic model. Accounting for more than 90% of 5-hydroxymethylcytosine peaks were distributed in the gene body in both the gastric cancer and control groups. The diagnostic model was developed based on five different 5-hydroxymethylcytosine modification genes, FBXL7, PDE3A, TPO, SNTG2 and STXBP5. The model could effectively distinguish gastric cancer patients from controls in the training (AUC = 0.95, sensitivity = 88.6%, specificity = 94.3%), validation (AUC = 0.87, sensitivity = 73.3%, specificity = 93.3%) and testing (AUC = 0.90, sensitivity = 81.9%, specificity = 90.2%) sets. The risk scores of the controls from the model were significantly lower than those of gastric cancer patients in bothour own data (P < 0.001) and GEO external testing data (P < 0.001), and no significant difference between different TNM stage patients (P = 0.09 and 0.66). Furthermore, there was no significant difference between the healthy control and benign gastric disease patients in the testing set from GEO (P = 0.10). The characteristics of 5-hydroxymethylcytosine in cell free DNA are specific to gastric cancer patients, and the diagnostic model constructed by five genes' 5-hydroxymethylcytosine features could effectively identify gastric cancer patients.

Abstract LB327: DNA methylation biomarkers for early bladder cancer detection and treatment response monitoring

Abstract Objectives: Bladder cancer (BC) stands as the 5th most prevalent cancer in the USA, with over 83,000 new cases diagnosed in 2023. The absence of non-invasive diagnostic tools for sensitive early BC detection and treatment response monitoring poses a significant challenge. This study evaluates the clinical potential of the Bladder CARE™ Assay for early BC detection and monitoring treatment response in BC patients. Methods: Under an institutional review board-approved protocol, voided urine samples were prospectively collected from USC patients with prior BC history under surveillance/anticancer treatment, prior genitourinary manipulation. Enrollment spanned February 2019 to September 2021. Samples underwent analysis using the Bladder CARE™ Assay, a DNA methylation test for quantitative BC and upper tract urothelial carcinoma (UTUC) detection from urine. Results were reported as Bladder CARE Index (BCI) and samples were categorized as “positive” (BCI &amp;gt; 5), “low-positive” (2.5 &amp;lt; BCI &amp;lt; 5), or “negative” (BCI &amp;lt; 2.5). Correlations between BCI value and categories, and clinicopathological findings were assessed. Results: A total of 110 previously diagnosed BC patients (median age: 74; 86% male) were enrolled in this study. Within 36 months post-TURBT, 24 patients (21.8%) showed evidence of recurrence. Bladder CARE™ Assay detected all recurrences, averagely 7.35 months earlier than cystoscopy. Of 55 patients (50%) undergoing anticancer therapies (45 BCG, 4 MMC, 2 GEM, 1 9UT, 1 GEM/DOCE, 1 MMC/BCG, and 1 BCG, MMC, and GEM), 7 (12.7%) recur (non-responders) and 11 (20%) did not show evidence of recurrence by 18 months post-TURBT (responders); 18-month post-operative data was unavailable for 37 patients (67.3%), which could not be classified in responders or non-responders. Bladder CARE™ detected 85.7% of non-responders, with BCI increasing prior to positive histology (avg. BCI: 86.1). In responders, BCI remained stable in negative/low-positive range post-TURBT (avg. BCI: 2.5). In addition, data from pre- and post-TURBT assessments were available for 20 of the 110 enrolled patients. The Bladder CARE™ Assay demonstrated a decrease in BCI post-TURBT in 19 of these 20 patients (95%). Conclusions: This prospective pilot study underscores the Bladder CARE™ Assay's capacity to pre-emptively detect BC months ahead of the gold standard. Furthermore, its quantitative nature offers prognostic insight, enabling non-invasive monitoring of patient response to anticancer treatments. A larger-scale study is the next step to validating these promising findings. Citation Format: Paolo Piatti, Sia Daneshmand, Sanam Ladi Seyedian, Saum Ghodossipour, Hamed Ahmadi, Suzanne Roberts, Alireza Ghoreifi, Michael Basin, Simin Hajian, Yap Ching Chew, Jeffrey Bhasin, Benjamin Jara, Lucy Sanossian, Hooman Djaladat, Anne Shuckman, Sumeet Bhavandia, Gangning Liang. DNA methylation biomarkers for early bladder cancer detection and treatment response monitoring [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB327.

Urine proteome profile of firefighters with exposure to emergency fire-induced smoke: A pilot study to identify potential carcinogenic effects

Firefighters are frequently exposed to a variety of chemicals formed from smoke, which pose a risk for numerous diseases, including cancer. Comparative urine proteome profiling could significantly improve our understanding of the early detection of potential cancer biomarkers. In this study, for the first time, we conducted a comparative protein profile analysis of 20 urine samples collected from ten real-life firefighters prior to and following emergency fire-induced smoke. Using a label-free quantitative proteomics platform, we identified and quantified 1325 unique protein groups, of which 45 proteins showed differential expressions in abundance in response to fire-smoke exposure (post) compared to the control (pre). Pathway analysis showed proteins associated with epithelium development (e.g., RHCG, HEG1, ADAMTSL2) and Alzheimer's disease (SORL1) were significantly increased in response to smoke exposure samples. A protein-protein-network study showed a possible link between these differentially abundant proteins and the known cancer gene (TP53). Moreover, a cross-comparison analysis revealed that seven proteins—ALDH1A1, APCS, POMC, COL2A1, RDX, DDAH2, and SDC4 overlapped with the previously published urine cancer proteome datasets, suggesting a potential cancer risk. Our findings demonstrated that the discovery proteomic platform is a promising analytical technique for identifying potential non-invasive biomarkers associated with fire-smoke exposure in firefighters that may be related to cancer.

Abstract LB108: Specific plasma DNA end-motifs distinguish patients with lung adenocarcinoma versus benign nodules

Abstract Background: Recent studies have identified blood plasma DNA as a rich source of biomarkers for non-invasive cancer detection. In particular, analysis of shorter DNA fragments or “fragmentomics” have shown promising results with numerous competing approaches such as those based on mutation calls, length distributions, or preferred fragmentation patterns. In this study, we assessed the potential of fragment end motif distributions in discriminating patients with lung adenocarcinoma versus benign tumors. Methods: Plasma samples from 37 patients (20 benign, 17 malignant) were subject to whole genome sequencing. For each sample, DNA fragments were grouped as short (30-99bp) and long (100-269bp) and end motif distributions were calculated by counting occurrences of 4-9bp sequences at either the 5’ or 3’ end of each fragment. A proprietary machine learning algorithm was employed to identify a minimal set of motifs capable of accurately distinguishing benign from malignant samples. Results: Just two 5’ end motifs in long fragments, CCAT and GACA, were sufficient for distinguishing malignant from benign samples in a two-step decision tree (89% accuracy with all samples, 86% accuracy in leave-one-out cross validation). Accuracy was significantly higher when compared to random guessing by shuffling labels (p=1.6*10−6, FWER &amp;lt; 0.05, Bonferroni correction, 351 tests). CCAT motif frequency was also significantly associated with cancer stage (p=0.04, Kruskal-Wallis test) and specifically elevated in early stage samples (I, II). Conclusions: These results suggest that plasma DNA end motif distributions are informative in discriminating patients with lung adenocarcinoma versus benign tumors. The two-motif model suggests that specific, rather than systemic changes in end motif distributions are more strongly associated with lung cancer patients and may point to specific mechanisms linked to cancer progression. Citation Format: Jason C. Hyun, Shun H. Yip, Neeti Swarup, Raghuraman Ramamurthy, Jordan C. Cheng, Irene Choi, Edmund Wong, Akanksha Arora, Denise Aberle, David T. Wong, Cheuk Y. Tang. Specific plasma DNA end-motifs distinguish patients with lung adenocarcinoma versus benign nodules [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB108.