McDonald et al. · Nature Communications · 2024

Space radiation damage rescued by inhibition of key spaceflight associated miRNAs

McDonald J. Tyson, Kim JangKeun, Farmerie Lily, Johnson Meghan L., Trovao Nidia S., Arif Shehbeel, Siew Keith, Tsoy Sergey, Bram Yaron, Park Jiwoon, Overbey Eliah, Ryon Krista, Haltom Jeffrey, Singh Urminder, Enguita Francisco J., Zaksas Victoria, Guarnieri Joseph W., Topper Michael, Wallace Douglas C., Meydan Cem, Baylin Stephen, Meller Robert, Muratani Masafumi, Porterfield D. Marshall, Kaufman Brett, Mori Marcelo A., Walsh Stephen B., Sigaudo-Roussel Dominique, Mebarek Saida, Bottini Massimo, Marquette Christophe A., Wurtele Eve Syrkin, Schwartz Robert E., Galeano Diego, Mason Christopher E., Grabham Peter, Beheshti Afshin

Read the paper The product used

The study

What was asked, and what was found

McDonald et al., Nature Communications, 2024 asked what changes inside a human blood vessel when three spaceflight associated microRNAs are blocked during exposure to deep space radiation. A three dimensional human umbilical vein endothelial cell microvessel model was irradiated with 0.5 Gy of simulated galactic cosmic radiation, and AUMantagomir oligonucleotides against miR-16-5p, miR-125b-5p and let-7a-5p were added to the medium at 0.5 μM each, a single dose 24 hours before irradiation. The paper names AUM BioTech as the supplier in the Methods.

Blocking any one of the three microRNAs preserved microvessel shape after irradiation, and the combination of all three was stronger. Treated cultures showed fewer 53BP1 DNA repair foci, back down to the level of unirradiated cultures, 90 minutes after exposure. RNA sequencing put the treated irradiated cultures closer to the unirradiated controls than to the irradiated ones. Twenty one genes were predicted targets of all three microRNAs and were restored by treatment, PCR was run on eight of those genes and seven agreed with the sequencing, with MSH5 the one that did not.

Pathway analysis showed that inflammatory response, tumour necrosis factor alpha, interferon and interleukin 6 signalling all rose with radiation and fell again with the antagomirs, as did induction of the NLRP3 inflammasome genes. Mitochondrial gene sets were widely affected, with 106 of 149 lowered when antagomirs were combined with irradiation. The same 21 genes moved in astronaut data from the NASA Twin Study, a JAXA mission and the Inspiration4 flight, which is how the authors tie the culture result back to people. Mice were used in this study only for microRNA sequencing after irradiation and received no antagomir.

Key findings

AUMantagomir oligonucleotides against any one of three microRNAs preserved the shape of irradiated human microvessels, and the three together worked better than any one alone.(Results, first section, Figures 1a and 1c)
Antagomir treatment cut the number of DNA double strand break repair foci back to the level of unirradiated cultures.(Results, DNA double strand break section, Figure 2b)
Whole transcriptome profiles of treated irradiated cultures sat closer to the unirradiated controls than to the irradiated ones.(Results, rescue section, Figure 4b)
Inflammatory signalling raised by radiation, including tumour necrosis factor alpha, interferon and interleukin 6 pathways, fell again when the antagomirs were added.(Results, pathway section, Figures 8a and 8b)
Radiation switched on the NLRP3 inflammasome genes and the antagomirs damped that induction.(Results, immune pathway section, Figure 9b)
The 21 genes rescued in culture also moved in astronaut samples from the NASA Twin Study, a JAXA mission and Inspiration4.(Results, astronaut section, Figure 6e)

Product: AUMantagomir
Targets: miR-16-5p · human · microRNAmiR-125b-5p · human · microRNAlet-7a-5p · human · microRNA
Models: mature human umbilical vein endothelial cell microvessels in collagen gel · 3D cell culturehuman umbilical vein endothelial cell angiogenesis model, antagomirs applied on days 2 and 3 · 3D cell cultureC57BL/6 female mice exposed to galactic cosmic radiation, used for microRNA sequencing only and not treated with antagomirs · in vivo
Delivery: a single dose added to the culture medium 24 hours before irradiation, with no transfection reagent described
Readouts: microvessel morphology by fluorescence imaging, 53BP1 DNA repair foci by immunocytochemistry, bulk RNA sequencing, RT-PCR validation, microRNA sequencing, gene set enrichment analysis
Measured: antagomir concentration in the 3D microvessel cultures: 0.5 μM each (single dose 24 h before irradiation, mature microvessel model)simulated deep space radiation dose: 0.5 Gy of galactic cosmic radiation (simplified five ion simulation at the NASA Space Radiation Laboratory)genes predicted to be simultaneous targets of all three microRNAs and restored by antagomir treatment: 21 genes (mirDIP target analysis of transcripts lowered by irradiation and restored by antagomirs)PCR check on the genes restored by antagomir treatment: 8 of the 21 tested, 7 agreeing with the sequencing (same isolated RNA as the sequencing; MSH5 was the exception, rising with 0.5 Gy and returning to the unirradiated level after treatment)time point at which DNA repair foci were counted: 90 min after irradiation (53BP1 immunocytochemistry in the microvessel cultures)mitochondrial gene sets significantly lowered when antagomirs were added to irradiation: 106 of 149 gene sets (mature microvessels, MitoPathways collection)metabolism linked mitochondrial gene sets downregulated by antagomirs with irradiation: 44 of 50 gene sets (mature microvessels)conservation of the three mature microRNAs between mouse and human: 100 % (mature sequences; precursor forms above 95%)experimentally validated gene targets among the predicted set: 10 of 10 for miR-16-5p, 13 of 14 for let-7a-5p and 9 of 15 for miR-125b-5p (cross referenced against miRTarBase, TarBase and starBase)
Figures: Fig. 1 Microvessel formation in the mature and angiogenesis models after irradiation, with and without each antagomir and the combination.Fig. 2 53BP1 DNA repair foci counted 90 minutes after irradiation, with and without antagomir treatment.Fig. 5 The 21 gene targets shared by the three microRNAs and returned to control levels by antagomir treatment.Fig. 10 Mitochondrial and integrated stress response pathways in the two culture models after irradiation with and without antagomirs.
Citation: McDonald et al. Space radiation damage rescued by inhibition of key spaceflight associated miRNAs. Nature Communications 2024;15:4825. doi:10.1038/s41467-024-48920-y

For research use only. Not for use in diagnostic or therapeutic procedures.