Spaceflight-Associated Vascular Remodeling and Gene Expression in Mouse Calvaria

Jamila H Siamwala; Brandon R Macias; Robert Healey; Brett Bennett; Alan R Hargens

doi:10.3389/fphys.2022.893025

Spaceflight-Associated Vascular Remodeling and Gene Expression in Mouse Calvaria

Front Physiol. 2022 May 13:13:893025. doi: 10.3389/fphys.2022.893025. eCollection 2022.

Authors

Jamila H Siamwala^{1

2

3}, Brandon R Macias^{1

4}, Robert Healey¹, Brett Bennett⁵, Alan R Hargens¹

Affiliations

¹ Department of Orthopedic Surgery, University of California, San Diego, San Diego, CA, United States.
² Department of Molecular Physiology, Pharmacology and Biotechnology, Brown University, Providence, RI, United States.
³ Vascular Research Laboratory, Providence Veterans Affairs Medical Center, Providence, RI, United States.
⁴ KBRwyle, Houston, TX, United States.
⁵ Association of Spaceflight Professionals, St. Petersburg, FL, United States.

Abstract

Astronauts suffer from a loss of bone mass at a rate of 1.5% per month from lower regions of the body during the course of long-duration (>30 days) spaceflight, a phenomenon that poses important risks for returning crew. Conversely, a gain in bone mass may occur in non-load bearing regions of the body as related to microgravity-induced cephalad fluid shift. Representing non-load bearing regions with mouse calvaria and leveraging the STS-131 (15-day) and BION-M1 (30-day) flights, we examined spatial and temporal calvarial vascular remodeling and gene expression related to microgravity exposure compared between spaceflight (SF) and ground control (GC) cohorts. We examined parasagittal capillary numbers and structures in calvaria from 16 to 23 week-old C57BL/6 female mice (GC, n = 4; SF, n = 5) from STS-131 and 19-20 week-old C57BL/6 male mice (GC, n = 6; SF, n = 6) from BION-M1 using a robust isolectin-IB4 vessel marker. We found that the vessel diameter reduces significantly in mice exposed to 15 days of spaceflight relative to control. Capillarization increases by 30% (SF vs. GC, p = 0.054) in SF mice compared to GC mice. The vessel numbers and diameter remain unchanged in BION-M1 mice calvarial section. We next analyzed the parietal pro-angiogenic (VEGFA) and pro-osteogenic gene (BMP-2, DMP1, RUNX2 and OCN) expression in BION-M1 mice using quantitative RT-PCR. VEGFA gene expression increased 15-fold while BMP-2 gene expression increased 11-fold in flight mice compared to GC. The linkage between vascular morphology and gene expression in the SF conditions suggests that angiogenesis may be important in the regulation of pathological bone growth in non-weight bearing regions of the body. Short-duration microgravity-mediated bone restructuring has implications in planning effective countermeasures for long-duration flights and extraterrestrial human habitation.

Keywords: BMP-2; VEGFA; angiogenesis; gene expression; long-duration flight; osteogenesis; spaceflight.