Abstract
Liquid biopsy of plasma is a simple and non-invasive technology that holds great promise in biomedical research. It is based on the analysis of nucleic acid-based biomarkers with predictive potential. In the present work, we have combined this concept with the FTA technology for sentinel mussels. We found that hemocytes collected from liquid biopsies can be readily fixed on FTA cards and used for long-term transcriptome analysis. We also showed that liquid biopsy is easily adaptable for metagenomic analysis of bacterial profiles of mussels. We finally provide evidence that liquid biopsies contained circulating cell-free DNA (ccfDNA) which can be used as an easily accessible genomic reservoir. Sampling of FTA-fixed circulating nucleic acids is stable at room temperature and does not necessitate a cold-chain protection. It showed comparable performance to frozen samples and is ideally adapted for sampling in remote areas, most notably in polar regions threatened by anthropogenic activities. From an ethical point of view, this minimally-invasive and non-lethal approach further reduces incidental mortality associated with conventional tissue sampling. This liquid biopsy-based approach should thus facilitate biobanking activities and development of omics-based biomarkers in mussels to assess the quality of aquatic ecosystems.
Highlights
Because of their ability to accumulate xenobiotics in their tissues, their wide distribution, and their ecological and economical importance, blue mussels (Mytilus spp.) have long been recognized as good biological indicators for monitoring the effects of pollution and climate change in marine habitats [1]
Isolation and analysis of RNA from hemocyte pellets fixed on Flinders Technology Associates (FTA) cards Because hemocytes are implicated in several physiological functions, such as phagocytosis of pathogens, homeostasis, and reproduction, there is increasing interest in studying how their transcriptome is modulated by environmental stress factors [24,25,26,27,28]
Other than a battery-operated centrifuge, it does not require specialized equipment and can be incorporated into a small, lightweight portable unit (S1 Fig). To test whether such sampling framework is compatible for multi-omics analysis of hemolymph samples, we first compared RNA yields obtained from frozen hemocytes and hemocytes that were fixed on FTA cards
Summary
Because of their ability to accumulate xenobiotics in their tissues, their wide distribution, and their ecological and economical importance, blue mussels (Mytilus spp.) have long been recognized as good biological indicators for monitoring the effects of pollution and climate change in marine habitats [1]. A relatively large number of biomarkers have been developed, often using a multi-tier approach. They include functional biomarkers based on enzymatic activities or cellular functions, or on measuring concentrations of specific stress indicators [2,3,4]. Measure of these biomarkers requires on-site analysis or tissue biopsy performed by trained personnel [5, 6]. It often requires an uninterrupted cold chain to maintain sample quality. The use of formalin-fixed tissue biopsy is an alternative,
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