Hypothetical LOC Genes as Biomarkers of Spaceflight Adaptation: A Comparative Study from ISS, Suborbital, and Earth-Based Experiments

Microgravity constitutes one of the most profound environmental stressors encountered by humans during spaceflight, capable of altering fundamental cellular processes and gene regulatory networks. While the effects of spaceflight on well-characterized protein-coding genes have been widely documented, little is known about the behavior of uncharacterized or poorly annotated genomic regions under these conditions. LOC (Locus) genes, often classified as long non-coding RNAs and excluded from conventional analyses, represent a largely unexplored component of the human transcriptome. In this study, we systematically investigated the transcriptional responses of LOC genes as part of the MESSAGE (Microgravity Associated Genetics) Science Mission, Turkiyes first human space biology initiative. Peripheral blood samples were collected from astronauts across five mission phases: pre-launch baseline, post-suborbital flight ([~]100 km), and on International Space Station (ISS) Days 4, 7, and 10 ([~]400 km). RNA-Seq analyses revealed six LOC genes with statistically significant expression changes (p < 0.05, Kruskal-Wallis test), alongside additional transcripts that, while not statistically significant, exhibited biologically meaningful temporal fluctuations. These dynamic profiles included continuous upregulation, transient activation with subsequent return to baseline, and delayed induction at later ISS stages, highlighting the functional diversity of LOC responses. To assess their translational potential, Open Reading Frame (ORF) analyses were performed on significant transcripts, revealing conserved ORF structures--most notably identical ORF33 sequences in LOC124905103 and LOC124900480-- suggesting coding capacity. Phylogenetic analyses further supported evolutionary clustering consistent with expression and ORF similarities. Collectively, these findings challenge the notion of LOC genes as transcriptional noise, instead positioning them as candidate biomarkers and functional elements of microgravity adaptation. By extending space biology research into the "dark genome," this study provides novel insights with potential implications for astronaut health monitoring and therapeutic development in long-duration missions.