Development Of Non-invasive Diagnostics Research Articles

Cervicovaginal Metabolome and Tumor Characteristics for Endometrial Cancer Detection and Risk Stratification.

Endometrial cancer is highly prevalent and lacking noninvasive diagnostic techniques. Diagnosis depends on histological investigation of biopsy samples. Serum biomarkers for endometrial cancer have lacked sensitivity and specificity. The objective of this study was to investigate the cervicovaginal environment to improve the understanding of metabolic reprogramming related to endometrial cancer and identify potential biomarker candidates for noninvasive diagnostic and prognostic tests. Cervicovaginal lavages were collected from 192 participants with endometrial cancer (n = 66) and non-malignant conditions (n = 108), and global untargeted metabolomics was performed. Using the metabolite data (n = 920), we completed a multivariate biomarker discovery analysis. We analyzed grade 1/2 endometrioid carcinoma (n = 53) and other endometrial cancer subtypes (n = 13) to identify shared and unique metabolic signatures between the subtypes. When compared to non-malignant conditions, downregulation of proline (P < 0.0001), tryptophan (P < 0.0001), and glutamate (P < 0.0001) was found among both endometrial cancer groups, relating to key hallmarks of cancer including immune suppression and redox balance. Upregulation (q < 0.05) of sphingolipids, fatty acids, and glycerophospholipids was observed in endometrial cancer in a type-specific manner. Furthermore, cervicovaginal metabolites related to tumor characteristics, including tumor size and myometrial invasion. Our findings provide insights into understanding the endometrial cancer metabolic landscape and improvement in diagnosis. The metabolic dysregulation described in this article linked specific metabolites and pathophysiological mechanisms including cellular proliferation, energy supply, and invasion of neighboring tissues. Furthermore, cervicovaginal metabolite levels related to tumor characteristics, which are used for risk stratification. Overall, development of noninvasive diagnostics can improve both the acceptability and accessibility of diagnosis.

Standard operating procedure to reveal prostate cancer specific volatile organic molecules by infrared spectroscopy

The growing number of prostate cancer cases is a real concern in modern society. Over 1.4 million new cases and about 400 thousand (>26%) deaths were registered worldwide in 2020 due to prostate cancer. The high mortality rate of prostate cancer is due to the lack of reliable early detection of the disease. Till now the most reliable diagnosis of cancer is tissue biopsy, which is an invasive process. A non-invasive or minimally invasive technique could lead to a diagnostic tool that will allow for saving or prolonging the lifespan of millions of lives. Metabolite-based diagnostics may have a better chance of early cancer detection. However, reliable detection techniques need to be developed. Infrared spectroscopy based gaseous-biofluid holds great promise towards the development of non-invasive diagnostics. A pilot study based on breath analysis by infrared spectroscopy showed promising results in distinguishing prostate cancer patients from healthy volunteers. Details of the spectral metabolic analysis are presented.

Breath volatile organic compounds for the gut-fatty liver axis: promise, peril, and path forward.

The worldwide interest in the gut microbiome and its impact on the upstream liver highlight a critical upside to breath research: it can uniquely measure otherwise unmeasurable biology. Bacteria make gases [volatile organic compounds (VOCs)] that are directly relevant to pathophysiology of the fatty liver and associated conditions, including obesity. Measurement of these VOCs and their metabolites in the exhaled breath, therefore, present an opportunity to safely and easily evaluate, on both a personal and a population level, some of our most pressing public health threats. This is an opportunity that must be pursued. To date, however, breath analysis remains a slowly evolving field which only occasionally impacts clinical research or patient care. One major obstacle to progress is that breath analysis is inherently and emphatically mutli-disciplinary: it connects engineering, chemistry, breath mechanics, biology and medicine. Unbalanced or incomplete teams may produce inconsistent and often unsatisfactory results. A second impediment is the lack of a well-known stepwise structure for the development of non-invasive diagnostics. As a result, the breath research landscape is replete with orphaned single-center pilot studies. Often, important hypotheses and key observations have not been pursued to maturation. This paper reviews the rationale and requirements for breath VOC research applied to the gut-fatty liver axis and offers some suggestions for future development.

Exosomal miRNAs as potential biomarkers of cardiovascular risk in children.

Intercellular interactions are essential for basic cellular activities and errors in either receiving or transferring these signals have shown to cause pathological conditions. These signals are not only regulated by membrane surface molecules but also by soluble secreted proteins, thereby allowing for an exquisite coordination of cell functions. Exosomes are released by cells upon fusion of multivesicular bodies (MVB) with the plasma membrane. Their envelope reflects their cellular origin and their surface and internal contents include important signaling components. Exosomes contain a wide variety of proteins, lipids, RNAs, non-transcribed RNAs, miRNAs and small RNAs that are representative to their cellular origin and shuttle from donor cells to recipient cells. The exosome formation cargo content and delivery is of immense biological interest because exosomes are believed to play major roles in various pathological conditions, and therefore provide unique opportunities for biomarker discovery and development of non-invasive diagnostics when examined in biological fluids such as urine and blood plasma. For example, circulating miRNAs in exosomes have been applied as functional biomarkers for diagnosis and outcomes prediction, while synthetic miRNAs in polymer-based nanoparticles are applicable for therapeutics. This review provides insights into the composition and functional properties of exosomes, and focuses on their potential value as diagnostic markers in the context of cardiovascular disease risk estimates in children who suffer from conditions associated with heightened prevalence of adverse cardiovascular disease, namely obesity and sleep-disordered-breathing.

Abstract LB-108: Exosome profiling of mRNAs and miRNAs in serum samples from GBM patients .

Abstract Background: Biomarker discovery and development in oncology has traditionally required the use of tissue, either FFPE or frozen. In recent years, the use of Circulating Tumor Cells (CTCs), cell free DNA (cfDNA), and most recently, exosomes, has allowed biomarker research in biofluids to evolve. Exosomes contain mRNA, microRNA, DNA and protein in a protected, highly stable vesicle. Recently, we have shown that large panels of mRNA genes can be run on small samples of frozen, archived serum. The ability to assess the expressions patterns in blood represents a potentially important development for biomarker research and the development of non-invasive diagnostics. Methods: Frozen, archived serum (2ml) was obtained from a glioblastoma clinical trial cohort, containing serum from both responders and non-responders to a novel therapy. We isolated the microvesicle fraction from the serum, extracted the RNA, and qualified the RNA using an Agilent 2100 Bioanalyzer on an Agilent 6000 RNA Picochip. A standard in-house control extraction was processed in parallel and subjected to quality control on the Bioanalyzer as well as RT-qPCR. The first cohort of samples (n = 60) were reverse transcribed to cDNA, and the cDNA was assayed on a human cancer mRNA panel Array by qPCR (Life Technologies).The cancer panel includes a total of 648 cancer related genes including endogenous controls. The second cohort of samples (n = 40) was reverse transcribed and analyzed by a miRNA panel of 758 different miRNA assays and controls. Results: Our data clearly demonstrate the ability to detect relevant gene expression patterns in microvesicle RNA isolated from patient serum. High quality RNA was extracted from the samples received, and an average of 80% of the 648 cancer associated mRNA genes and 36% of the miRNAs were detected in the samples with the volumes used in this study. Data was demonstrated to be reproducible between and among samples and suggests a biological pattern of RNAs in serum exosomes that could be used to monitor response to treatment. Conclusions: RNA isolated from the microvesicle fraction of serum contains abundant, high-quality RNA, including mRNA. The analysis of a large panel of genes from a small sample of serum is possible, demonstrating that RNA isolated from microvesicles from peripheral blood has the potential to be used as a source of genetic material for non-invasive diagnostic tests. Citation Format: Johan Skog, Stefan Bentink, Lori LoGuidice, Charlotte Romain, Susan Belzer, Mikkel Noerholm. Exosome profiling of mRNAs and miRNAs in serum samples from GBM patients . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-108. doi:10.1158/1538-7445.AM2013-LB-108