Dr. Yan Lin, Chief Physician of the Department of Medical Imaging at the Second Affiliated Hospital of Shantou University Medical College, and her research team, in collaboration with partners, have developed and optimized a set of fluid metabolite markers based on nuclear magnetic resonance (NMR) technology. This breakthrough holds promising potential as a powerful tool for early diagnosis and screening of esophageal squamous cell carcinoma (ESCC). Supported by grants from the National Natural Science Foundation of China and the Natural Science Foundation of Guangdong Province, their findings were published on March 19th in Nature Communications.
Research Overall Design Concept: Visual from Interviewee
"We only need five serum or urine metabolites to efficiently identify early-stage esophageal squamous cell carcinoma, greatly improving the feasibility of early screening," said Lin Yan, the project leader and corresponding author of the paper, to the China Science Daily. Currently, such nuclear magnetic resonance metabolomics detection methods have been clinically applied in countries like the UK and Canada. However, this research represents the first large-scale application of these methods for esophageal squamous cell carcinoma sample detection.
Reviewer evaluations of this research stated that it "represents the largest and most comprehensive metabolomics analysis of esophageal squamous cell carcinoma to date" and "demonstrates tremendous effort and provides a valuable resource for the scientific community."
Esophageal cancer is predominantly squamous cell carcinoma in China, unlike adenocarcinoma which is more common in Western countries. Esophageal squamous cell carcinoma is the most common subtype of esophageal cancer in China, and understanding its metabolic phenotype changes before malignant histological alterations may provide important opportunities for early detection and timely intervention.
Predictive statistics suggest that by 2040, the global incidence of newly diagnosed esophageal squamous cell carcinoma cases could reach as high as 806,000. However, existing clinical early detection methods have limitations, leading many patients to miss the optimal treatment window. In China, based on health economic analysis, early screening for esophageal squamous cell carcinoma has been proven to be one of the most cost-effective cancer prevention and control strategies.
Yet, the latest "Guidelines for Screening, Early Diagnosis, and Early Treatment of Esophageal Cancer in China" still show that due to insufficient evidence, there are currently no recommended biomarkers for screening or diagnosing esophageal cancer. Therefore, there is an urgent need to develop a reliable, non-invasive, accessible, and cost-effective tool to promote early detection of esophageal squamous cell carcinoma.
"Our goal is to discover tissue-specific metabolic biomarkers of esophageal squamous cell carcinoma progression and, based on these markers, to construct and optimize fluid metabolism classifiers based on nuclear magnetic resonance technology," explained Lin Yan. The team adopted a comprehensive research strategy, collecting 1,153 multidimensional matched tissue, serum, and urine samples, utilizing nuclear magnetic resonance and targeted mass spectrometry technologies for cross-platform detection, and ensuring the reliability of the results through multi-center validation.
Eight Years of Effort
Lin Yan told the China Science Daily that the large sample of esophageal squamous cell carcinoma in the study has been collected, tested, and analyzed since 2016, spanning 8 years, and has brought together team efforts and support from multiple clinical centers. Establishing a rigorously standardized biobank, conducting rigorous clinical trial registration, continuously optimizing sample pretreatment schemes and detection parameters based on practical considerations, interpreting massive high-dimensional data, and strictly adhering to the requirements of the "Standards for Reporting of Diagnostic Accuracy Studies" to produce standardized reports on research results, all reflect the result of efforts.
Lin Yan has 18 years of research experience in detecting body metabolites using ultra-high-field 1H-NMR. She pursued her PhD at the University of Nottingham's Sir Peter Mansfield Magnetic Resonance Centre from 2008 to 2011, under the supervision of Professor Peter Morris, conducting research on detecting human metabolites using 7T 1H-NMR. After completing her doctorate, she continued her research on detecting metabolites using ultra-high field strength 1H-NMR upon returning to China.
"Through comprehensive review and analysis of published literature, we noticed that most existing metabolomics studies on esophageal squamous cell carcinoma focus on exploring fluid samples such as serum or urine. However, it cannot be ignored that various potential factors such as environment, lifestyle, phenotype differences, and comorbidities may affect fluid metabolism, leading to certain differences between the fluid metabolism profile and characteristic molecular events in esophageal squamous cell carcinoma tissue," said Lin Yan.
Based on tissue metabolic characteristics confirmed by both nuclear magnetic resonance and targeted mass spectrometry technologies, Lin Yan's team, in collaboration with researchers from Shantou University Medical College Affiliated Tumor Hospital, Shantou Central Hospital, the Swiss Federal Institute of Technology Lausanne, and Chongqing University Affiliated Tumor Hospital, developed and optimized a set of 16 serum metabolite markers and 10 urine metabolite markers based on nuclear magnetic resonance technology. These fluid markers not only accurately reflect the unique metabolic characteristics of esophageal squamous cell carcinoma tissue but also have sufficient clinical sensitivity.
Furthermore, by further simplifying the generated fluid metabolic marker combinations based on nuclear magnetic resonance, even including only five arbitrary types of serum or urine metabolite markers, effective diagnosis and prediction of early-stage esophageal squamous cell carcinoma can be achieved. This makes it applicable in clinical screening and potentially provides a practical model with wide applicability for other research institutions.
Aiding Early Screening of Esophageal Squamous Cell Carcinoma
Co-corresponding author and professor at the Second Affiliated Hospital of Shantou University Medical College, Wu Renhua, stated that preliminary research has established a tissue biomarker resource library for digestive tract tumors, confirming the good performance of fluid metabolic fingerprint spectra based on nuclear magnetic resonance technology in distinguishing digestive tract tumor patients from healthy individuals, indicating the potential application value of fluid metabolic fingerprint spectra in esophageal squamous cell carcinoma detection.
Using proton nuclear magnetic resonance as the driving tool for research, combined with targeted quantitative mass spectrometry technology and machine learning algorithms, researchers conducted cross-platform detection and analysis of 1,153 multidimensional matched samples. They discovered significant disruptions in the "alanine, aspartate, and glutamate metabolism pathway" during the development of esophageal squamous cell carcinoma. This finding not only reveals the metabolic characteristics of esophageal squamous cell carcinoma but also provides important clues for subsequent selection of early diagnostic markers. Dr. Ma Changchun, co-first author of the paper and Associate Chief Physician at the Affiliated Tumor Hospital of Shantou University Medical College, pointed out that their research revealed changes in the "alanine, aspartate, and glutamate metabolism" pathway at both tissue and fluid levels throughout the progression of esophageal squamous cell carcinoma (ESCC), from normal mucosa to early and late stages of the disease, as well as from tumor burden to recovery after tumor removal until reaching a healthy state. This suggests that this metabolic pathway is a noteworthy characteristic molecular event in the progression of ESCC.
Zhao Yan, co-first author of the paper and a Ph.D. student from the 2021 cohort at Shantou University Medical College, mentioned to Chinese Science Bulletin that considering tissue-specific biomarkers detectable in bodily fluids have been identified, the next step is to promptly validate these findings in high-risk populations including those with low- and high-grade intraepithelial neoplasia, gastroesophageal reflux disease, and Barrett's esophagus, among others, to further refine the model. Additionally, the team will delve into multidisciplinary integration analyses including metabolomics, proteomics, and metagenomics to elucidate the origin and functional mechanisms of metabolites, with relevant work currently underway.
"This study primarily focuses on the Southern Chinese population, and its generalizability to other regions requires further exploration. At present, the sample size in the early stages of the study is limited, hence it's essential to conduct larger-scale prospective cohort studies in different regions and rigorously control confounding factors to validate our findings," emphasized Lin Yan.
It's noted that the research findings, with Lin Yan as the first inventor, have secured three national invention patents and have been validated through multicenter samples. This not only offers new insights and methods for the early diagnosis of ESCC but also accumulates valuable experience for the team in the fields of medical imaging and tumor diagnosis.
For more information about the related paper, please visit: https://doi.org/10.1038/s41467-024-46837-0
