Interactome of miRNAs and transcriptome of human umbilical cord endothelial cells exposed to short-term simulated microgravity

Abstract

Adaptation of humans in low gravity conditions is a matter of utmost importance when efforts are on to a gigantic leap in human space expeditions for tourism and formation of space colonies. In this connection, cardiovascular adaptation in low gravity is a critical component of human space exploration. Deep high-throughput sequencing approach allowed us to analyze the miRNA and mRNA expression profiles in human umbilical cord vein endothelial cells (HUVEC), cultured under gravity (G), and stimulated microgravity (MG) achieved with a clinostat. The present study identified totally 1870 miRNAs differentially expressed in HUVEC under MG condition when compared to the cells subjected to unitary G conditions. The functional association of identified miRNAs targeting specific mRNAs revealed that miRNAs, hsa-mir-496, hsa-mir-151a, hsa-miR-296-3p, hsa-mir-148a, hsa-miR-365b-5p, hsa-miR-3687, hsa-mir-454, hsa-miR-155-5p, and hsa-miR-145-5p differentially regulated the genes involved in cell adhesion, angiogenesis, cell cycle, JAK-STAT signaling, MAPK signaling, nitric oxide signaling, VEGF signaling, and wound healing pathways. Further, the q-PCR based experimental studies of upregulated and downregulated miRNA and mRNAs demonstrate that the above reported miRNAs influence the cell proliferation and vascular functions of the HUVEC in MG conditions effectively. Consensus on the interactome results indicates restricted fluctuations in the transcriptome of the HUVEC exposed to short-term MG that could lead to higher levels of endothelial functions like angiogenesis and vascular patterning.

The Impact of Spaceflight and Microgravity on the Human Islet-1+ Cardiovascular Progenitor Cell Transcriptome

“…These included TGF-β signaling. Similarly, a group evaluating the effect of simulated microgravity on human umbilical cord endothelial cells found that microgravity altered the expression of miRNAs involved in cell proliferation, cell cycle regulation, TGF-β signaling, VEGF signaling, MAPK signaling, Jak-STAT signaling, focal adhesion, and wound healing [73].…”

Effect of Microgravity on Endothelial Cell Function, Angiogenesis, and Vessel Remodeling During Wound Healing

“…While the events and mechanisms controlling wound healing are well known and characterized, the pharmacological interventions to prevent or treat healing dysfunction are few and nowadays still under evaluation and validation on Earth. The data available in relation to unloading conditions document that the impaired wound healing results from the following mechanisms: 1) persistent inflammation with neutrophil infiltration ( Dovi et al, 2003 ; Radek et al, 2008 ) and overall alteration of the inflammatory phase; 2) altered blood flow with more permeable vessels ( McDonald et al, 1992 ), presumably linked to increased NO and VEGF levels ( Shi et al, 2012 ; Dittrich et al, 2018 ); 3) an altered neovascularization that may result from impaired EC proliferation and migration in response to angiogenic factors, increased EC apoptosis, and altered gene expression and signalling pathways ( Morbidelli et al, 2005 ; Radek et al, 2008 ; Li et al, 2018 ; Kasiviswanathan et al, 2020 ); 4) exposure to simulated microgravity also resulting in enhanced ROS production that may contribute to unloading-induced oxidative stress. The systemic oxidative status can be derived from radiation-induced immune system alterations especially relevant in long-duration space flight ( Rizzo et al, 2012 ).…”

“…Examples are tissue engineering, cocultures, 3D multicellular structures, lab-on-chip approaches ( Grimm et al, 2014 ; Ma et al, 2014 ; Huang et al, 2020 ), which are starting to substitute experimental animals due to ethical issues. Thorough comprehension of the molecular and biochemical mechanisms underlying cellular responses is coming from omics techniques and RNA sequence analysis of samples from simulated and real microgravity experiments ( Ma et al, 2014 ; Mao et al, 2018 ; Cui et al, 2020 ; Kasiviswanathan et al, 2020 ). The expectations from these types of experiments are high.…”

“…Recently, in human ECs cultured under simulated microgravity achieved with a clinostat, a total of 1,870 miRNAs were found to be differentially expressed with respect to normal gravity. The functional association of identified miRNAs targeting specific mRNAs revealed that a series of miRNAs (hsa-mir-496, hsa-mir-151a, hsa-miR-296-3p, hsa-mir-148a, hsa-miR-365b-5p, hsa-miR-3687, hsa-mir-454, hsa-miR-155-5p, and hsa-miR-145-5p) differentially regulated the genes involved in cell adhesion, angiogenesis, cell cycle, JAK-STAT signaling, MAPK signaling, NO signaling, and VEGF signaling, finally favoring angiogenesis ( Kasiviswanathan et al, 2020 ). In a study on endothelial progenitor cells, cultured in simulated microgravity, a facilitation of functional angiogenic properties has been reported, with increased HIF-1α and eNOS/NO induced FAK/ERK1/2 pathway upregulation in HUVEC ( Kong et al, 2021 ).…”

Effect of Space Flight on the Behavior of Human Retinal Pigment Epithelial ARPE-19 Cells and Evaluation of Coenzyme Q10 Treatment

“…Some microRNAs we identified to be activated in ARPE-19 cells cultured on board ISS are involved in cell response to spaceflight. Among them, hsa-miR-296 expression is affected in HUVEC cultured in a 3D clinostat to simulate microgravity, and it influences cell proliferation and vascular function in microgravity conditions [79]. hsa-miR-494 is upregulated in C2C12 myoblast cells subjected to clinorotation conditions and in osteoblasts isolated from tailsuspended rats, and this upregulation correlates with inhibition of osteoclast differentiation [80].…”