Extracellular RNAs present in biofluids have emerged as potential biomarkers for disease. Where most studies focus on blood-derived fluids, other biofluids may be more informative. We present an atlas of messenger, circular, and small RNA transcriptomes of a comprehensive collection of 20 human biofluids. By means of synthetic spike-in controls, we compare RNA content across biofluids, revealing a 10,000-fold difference in concentration. The circular RNA fraction is increased in most biofluids compared to tissues. Each biofluid transcriptome is enriched for RNA molecules derived from specific tissues and cell types. Our atlas enables an informed selection of the most relevant biofluid to monitor particular diseases. To verify the biomarker potential in these biofluids, four validation cohorts representing a broad spectrum of diseases were profiled, revealing numerous differential RNAs between case and control subjects. Spike-normalized data are publicly available in the R2 web portal for further exploration.

Austrian Institute of Technology Center for Health and Bioresources Molecular Diagnostics Vienna Austria

Center for Medical Genetics Department of Biomolecular Medicine Ghent University C Heymanslaan 10 9000 Ghent Belgium

Central European Institute of Technology Masaryk University Brno Czech Republic

Department of Dermatology Ghent University Hospital C Heymanslaan 10 9000 Ghent Belgium

Department of Gastroenterology Ghent University Hospital C Heymanslaan 10 9000 Ghent Belgium

Department of Internal Medicine 3 Clinical Division of Endocrinology and Metabolism Unit of Gender Medicine Medical University of Vienna Vienna Austria

Department of Neurology Ghent University Hospital C Heymanslaan 10 9000 Ghent Belgium

Department of Obstetrics Women's Clinic Ghent University Hospital C Heymanslaan 10 9000 Ghent Belgium

Department of Oncogenomics Amsterdam University Medical Centers University of Amsterdam Amsterdam the Netherlands

Department of Ophthalmology Ghent University Hospital C Heymanslaan 10 9000 Ghent Belgium

Department of Pathology University Hospital Brno Brno Czech Republic

Department of Reproductive Medicine Ghent University Hospital C Heymanslaan 10 9000 Ghent Belgium

Department of Respiratory Medicine Ghent University Hospital C Heymanslaan 10 9000 Ghent Belgium

Department of Rheumatology Ghent University Hospital C Heymanslaan 10 9000 Ghent Belgium

Helmholtz Zentrum München German Research Center for Environmental Health Institute of Computational Biology Neuherberg Germany

Illumina San Diego CA USA

Masaryk Memorial Cancer Institute Department of Comprehensive Cancer Care Brno Czech Republic

OncoRNALab Cancer Research Institute Ghent Ghent University C Heymanslaan 10 9000 Ghent Belgium

University of Texas Health Science Center School of Biomedical Informatics Center for Precision Health Houston TX USA

VIB Inflammation Research Center Ghent University Technologiepark 71 9052 Ghent Belgium

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$a Charting Extracellular Transcriptomes in The Human Biofluid RNA Atlas / $c E. Hulstaert, A. Morlion, F. Avila Cobos, K. Verniers, J. Nuytens, E. Vanden Eynde, N. Yigit, J. Anckaert, A. Geerts, P. Hindryckx, P. Jacques, G. Brusselle, KR. Bracke, T. Maes, T. Malfait, T. Derveaux, V. Ninclaus, C. Van Cauwenbergh, K. Roelens, E. Roets, D. Hemelsoet, K. Tilleman, L. Brochez, S. Kuersten, LM. Simon, S. Karg, A. Kautzky-Willers, M. Leutner, C. Nöhammer, O. Slaby, RW. Prins, J. Koster, S. Lefever, GP. Schroth, J. Vandesompele, P. Mestdagh

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$a Extracellular RNAs present in biofluids have emerged as potential biomarkers for disease. Where most studies focus on blood-derived fluids, other biofluids may be more informative. We present an atlas of messenger, circular, and small RNA transcriptomes of a comprehensive collection of 20 human biofluids. By means of synthetic spike-in controls, we compare RNA content across biofluids, revealing a 10,000-fold difference in concentration. The circular RNA fraction is increased in most biofluids compared to tissues. Each biofluid transcriptome is enriched for RNA molecules derived from specific tissues and cell types. Our atlas enables an informed selection of the most relevant biofluid to monitor particular diseases. To verify the biomarker potential in these biofluids, four validation cohorts representing a broad spectrum of diseases were profiled, revealing numerous differential RNAs between case and control subjects. Spike-normalized data are publicly available in the R2 web portal for further exploration.

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