Secretome profiling reveals acute changes in oxidative stress, brain homeostasis, and coagulation following short-duration spaceflight

Abstract

As spaceflight becomes more common with commercial crews, blood-based measures of crew health can guide both astronaut biomedicine and countermeasures. By profiling plasma proteins, metabolites, and extracellular vesicles/particles (EVPs) from the SpaceX Inspiration4 crew, we generated “spaceflight secretome profiles,” which showed significant differences in coagulation, oxidative stress, and brain-enriched proteins. While >93% of differentially abundant proteins (DAPs) in vesicles and metabolites recovered within six months, the majority (73%) of plasma DAPs were still perturbed post-flight. Moreover, these proteomic alterations correlated better with peripheral blood mononuclear cells than whole blood, suggesting that immune cells contribute more DAPs than erythrocytes. Finally, to discern possible mechanisms leading to brain-enriched protein detection and blood-brain barrier (BBB) disruption, we examined protein changes in dissected brains of spaceflight mice, which showed increases in PECAM-1, a marker of BBB integrity. These data highlight how even short-duration spaceflight can disrupt human and murine physiology and identify spaceflight biomarkers that can guide countermeasure development.

Targeted proteomic analysis of dry blood spot samples collected during long-term spaceflights

“…Quantitative analysis of blood proteins is not an easy task, and in-depth analysis of a wide panel of proteins using mass-spectrometric methods requires additional efforts and should rely on well-validated reproducible protocols [11,14,18,19]. There are several strategies that are recommended for blood proteomics and clinical studies.…”

“…There are several strategies that are recommended for blood proteomics and clinical studies. We followed protocols and recommendations for absolute concentration measurements of more than 200 proteins in plasma/serum and DBS which was proposed by the Prof. Borcher's team [11,18,19]. Finally, 119 proteins were quanti ed by LC-MRM MS in ≥ 2/3 of 58 DBS samples collected from 7 Russian cosmonauts during long-duration SF.…”

“…Our group has previously demonstrated changes in the proteome composition after long-duration (169-199 days) missions on the ISS, but only samples collected before and after SF were analyzed [17,18]. As space ight becomes more common with commercial crews, blood-based measures become actual for health monitoring during shortduration space ight and the rst study already performed within the SpaceX Inspiration4 (i4) mission [19] The aim of the current study was to apply the method of blood proteome analysis with DBS microsampling technique to study the physiological response to SF conditions at the molecular level. Targeted mass-spectrometry based proteomics with a validated assay of stable isotope-labeled peptide standard (SIS) for 119 blood proteins was selected as a robust and precise tool for quantitative analysis.…”