Cited by 93Spaceflight effects on biologyBone Metabolism and DiseasesAutophagy in Disease and Therapy

Microgravity control of autophagy modulates osteoclastogenesis

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Sambandam, Yuvaraj, Townsend, Molly T., Pierce, Jason J., Lipman, Cecilia M., Haque, Azizul, Bateman, Ted A., Reddy, Sakamuri V.·2014·Bone

Abstract

Evidence indicates that astronauts experience significant bone loss during space mission. Recently, we used the NASA developed rotary cell culture system (RCCS) to simulate microgravity (μXg) conditions and demonstrated increased osteoclastogenesis in mouse bone marrow cultures. Autophagy is a cellular recycling process of nutrients. Therefore, we hypothesize that μXg control of autophagy modulates osteoclastogenesis. Real-time PCR analysis of total RNA isolated from mouse bone marrow derived non-adherent cells subjected to modeled μXg showed a significant increase in autophagic marker Atg5, LC3 and Atg16L mRNA expression compared to ground based control (Xg) cultures. Western blot analysis of total cell lysates identified an 8.0-fold and 7.0-fold increase in the Atg5 and LC3-II expression, respectively. Confocal microscopy demonstrated an increased autophagosome formation in μXg subjected RAW 264.7 preosteoclast cells. RT2 profiler PCR array screening for autophagy related genes identified that μXg upregulates intracellular signaling molecules associated with autophagy, autophagosome components and inflammatory cytokines/growth factors which coregulate autophagy in RAW 264.7 preosteoclast cells. Autophagy inhibitor, 3-methyladenine (3-MA) treatment of mouse bone marrow derived non-adherent mononuclear cells showed a significant decrease in μXg induced Atg5 and LC3 mRNA expression in the presence or absence of RANK ligand (RANKL) stimulation. Furthermore, RANKL treatment significantly increased (8-fold) p-CREB transcription factor levels under μXg as compared to Xg cultures and 3-MA inhibited RANKL increased p-CREB expression in these cells. Also, 3-MA suppresses μXg elevated osteoclast differentiation in mouse bone marrow cultures. Thus, our results suggest that μXg induced autophagy plays an important role in enhanced osteoclast differentiation and could be a potential therapeutic target to prevent bone loss in astronauts during space flight missions.

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Crossref · 10.1016/j.bone.2014.01.004

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  • Microgravity produced multiple autophagosomes in RAW 264.7 cultures.

    “produced multiple autophagosomes in RAW 264.7 cultures”

    From Osteoclasts and Microgravity (2020) · cites this paper as [67]

  • 3-MA inhibition of autophagy in RANKL/CSF+ marrow cultures inhibited osteoclastogenesis.

    “3-MA inhibition of autophagy in the RANKL/CSF+ marrow cultures inhibited osteoclastogenesis”

    From Osteoclasts and Microgravity (2020) · cites this paper as [67]

  • Autophagy plays an important role in enhanced osteoclast differentiation.

    “autophagy plays an important role in enhanced osteoclast differentiation”

    From Osteoclasts and Microgravity (2020) · cites this paper as [67]

  • Microgravity increased the expression of autophagic genes in RAW 264.7 cultures.

    “microgravity increased the expression of autophagic genes”

    From Osteoclasts and Microgravity (2020) · cites this paper as [67]

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Authors
Sambandam, Y., Townsend, M. T., Pierce, J. J., Lipman, C. M., Haque, A., Bateman, T. A., & Reddy, S. V.
Affiliations
  • Medical University of South Carolina (US)
  • University of North Carolina at Chapel Hill (US)
Corresponding author
Reddy, Sakamuri V.
Venue
Bone
Publisher
Elsevier BV
Published
2014-04
Volume
61
Pages
125-131
ISSN
  • 1873-2763 (ISSN-L)
  • 8756-3282
OpenAlex
W2078536317
Cited by
93
References
46
Citations by year
2026: 4 · 2025: 5 · 2024: 4 · 2023: 3 · 2022: 3 · 2021: 10 · 2020: 20 · 2019: 8 · 2018: 12 · 2017: 5 · 2016: 8 · 2015: 7 · 2014: 3 · 2012: 1
FWCI
5.10
Type
journal article
Language
en
Open access
Locations
  • Bone (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
  • UNC Libraries
Funders
  • South Carolina EPSCoR
  • National Institute of Arthritis and Musculoskeletal and Skin Diseases (R01AR059221)
  • Office of Experimental Program to Stimulate Competitive Research
Topics
  • Health Sciences › Medicine › Physiology › Spaceflight effects on biology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bone Metabolism and Diseases
  • Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
Keywords
microgravity, autophagy, osteoclastogenesis, spaceflight, rotary cell culture system, ATG5, LC3, RANK ligand, CREB phosphorylation, 3-methyladenine, autophagy inhibition, bone marrow mononuclear cells, osteoporosis prevention
MeSH terms
Animals, Autophagy, Bone Marrow Cells, Bone Resorption, Cell Differentiation, Cells, Cultured, Mice, Inbred C57BL, Osteoclasts, Stem Cells, Weightlessness, Blotting, Western, Weightlessness Simulation, Microscopy, Confocal, Mice, Real-Time Polymerase Chain Reaction

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