This study examined the effects of exposure to low-Earth orbit (LEO) on hematopoietic stem and progenitor cell function, gene expression, telomere length, mutation burden, and <a href="https://www.ncbi.nlm.nih.gov/datasets/gene/103/">ADAR1</a> isoform expression, evaluated 32-45 days post-spaceflight compared to ground control in 3D nanobioreactors. Enriched <a href="https://www.ncbi.nlm.nih.gov/datasets/gene/947/">CD34+</a> hematopoietic stem and progenitor cells (HSPCs) derived from human normal bone marrow were transduced with a FUCCI2BL cell cycle reporter, and seeded into twin niche nanobioreactors for month-long culture on either the International Space Station (spaceflight) or at Kennedy Space Center (ground control) as part of NASA SpaceX Commercial Resupply Service missions (SpX-CRS24, SpX-CRS25, SpX-CRS26, and SpX-CRS27). Upon mission completion, live cells were flushed from the nanobioreactors and subjected to downstream functional clonogenic assays and multi-omic analysis. Whole transcriptome sequencing was performed at The Scripps Research Institute Next Generation Sequencing Core on Illumina NextSeq 2000 sequencer. Whole genomic sequencing was performed by Novogene Corporation on Illumina NovaSeq X Plus sequencer. Single cell RNA sequencing was performed at The Salk Institute for Biological Studies on Illumina NovaSeq X Plus sequencer. Post-spaceflight analysis revealed increased C-to-T and clonal hematopoiesis-associated mutations, repetitive element and base deaminase deregulation, decreased telomere maintenance gene expression, and decreased self-renewal capacity compared to ground controls. These results demonstrate the space-associated aging effects in HSPCs after month-long exposure to LEO, providing insights into utilizing the LEO environment to model space-accelerated aging and other age-related malignancies.

Nanobioreactor Detection of Space-Associated Hematopoietic Stem and Progenitor Cell Aging