Microgravity control of autophagy modulates osteoclastogenesis
Open accessAbstract
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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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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Self-citationRoles of autophagy in orthodontic tooth movement
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Rivera, M. V., Cornejo, J., Huallpayunca, K., Diaz, A. B., Ortiz‐Benique, Z. N., Reina, A. D., Jamanca-Lino, G., & Ticllacuri, V. (2020) · Revista de la Facultad de Medicina Humana · doi:10.25176/rfmh.v20i2.2920 · Cited by 27 · see citing works
“En dicho proceso está involucrada la autofagia, con niveles acentuados de marcadores de genes (Atg5, LC3 y Atg16L), además el aumento de la expresión de la fosfolipasa PLCγ2, moléculas de señalización de proteínas quinasas activadas por mitógenos MAPK/ERK, P38 y activación de osteoclastos a través del receptor activad…”
BackgroundZhu, L., Nie, L., Xie, S., Li, M., Zhu, C., Qiu, X., Kuang, J., Liu, C., Lu, C., Li, W., et al. (2021) · Cell Reports · doi:10.1016/j.celrep.2020.108600 · Cited by 26 · see citing works
Haque, A., Drasites, K. P., Cox, A., Capone, M., Myatich, A., Shams, R., Matzelle, D. D., Garner, D. P., Bredikhin, M., Shields, D. C., et al. (2021) · Neurochemical Research · doi:10.1007/s11064-021-03401-2 · Cited by 26 · see citing works
“7 macrophage cells [58] were cultured in complete DMEM (Dulbecco’s modified Eagle’s medium) with high”
MethodSelf-citationPhysical activity, nutrition, and bone health
Amato, A., Baldassano, S., Cortis, C., Cooper, J., & Proia, P. (2018) · Human Movement · doi:10.5114/hm.2018.77318 · Cited by 23 · see citing works
Diamagnetic Levitation Promotes Osteoclast Differentiation From RAW264.7 Cells
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“ditions is of great importance as a promising way to elucidate the detailed mechanism underlying microgravity-induced bone loss [24], [30].”
CKIP-1 knockout offsets osteoporosis induced by simulated microgravity
Zhang, X., Wang, Q., Wan, Z., Li, J., Liu, L., & Zhang, X. (2016) · Progress in Biophysics and Molecular Biology · doi:10.1016/j.pbiomolbio.2016.09.007 · Cited by 22 · see citing works
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Autophagy: A Promising Target for Age-related Osteoporosis
Wang, T., He, H., Liu, S., Jia, C., Fan, Z., Zhong, C., Yu, J., Liu, H., & He, C. (2018) · Current Drug Targets · doi:10.2174/1389450119666180626120852 · Cited by 22 · see citing works
Simulated microgravity significantly altered metabolism in epidermal stem cells
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Mechanical protein polycystin-1 directly regulates osteoclastogenesis and bone resorption
Huang, M., Zhou, J., Li, X., Liu, R., Jiang, Y., Chen, K., Jiao, Y., Yin, X., Liu, L., Sun, Y., et al. (2024) · 中国科学通报:英文版 · doi:10.1016/j.scib.2024.04.044 · Cited by 20 · see citing works
“Recently, several studies have reported that osteoclastic lineage cells can directly sense mechanical stress [12,16,47,48].”
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Orthodontic loading activates cell-specific autophagy in a force-dependent manner
Jacox, L. A., Tang, N., Li, Y., Bocklage, C., Graves, C. L., Coats, S., Miao, M. Z., Glesener, T., Kwon, J., Giduz, N., et al. (2022) · American Journal of Orthodontics and Dentofacial Orthopedics · doi:10.1016/j.ajodo.2020.09.034 · Cited by 20 · see citing works
Huyan, T., Du, Y., Dong, D., Li, Q., Zhang, R., Yang, J., Yang, Z., Li, J., & Shang, P. (2018) · Acta Astronautica · doi:10.1016/j.actaastro.2018.08.044 · Cited by 19 · see citing works
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“Mitf, a dimeric transcription factor involved in autophagy and lysosomal biogenesis, has been reported to be suppressed by Mul-A in melanocytes [37, 38].”
Andreazzoli, M., Angeloni, D., Broccoli, V., & Demontis, G. C. (2017) · Frontiers in Astronomy and Space Sciences · doi:10.3389/fspas.2017.00002 · Cited by 15 · see citing works
“A role for the autophagic control of osteoclastogenesis in response to microgravity has been proposed for mouse bone (Sambandam et al., 2014).”
BackgroundEpigenetic Regulation of Autophagy in Bone Metabolism
Zhang, Y., Wang, Q., Xue, H., Guo, Y., Wei, S., Li, F., Gong, L., Pan, W., & Jiang, P. (2024) · Function · doi:10.1093/function/zqae004 · Cited by 14 · see citing works
Anti-osteoclastogenesis of Mineral Trioxide Aggregate through Inhibition of the Autophagic Pathway
Cheng, X., Zhu, L., Zhang, J., Yu, J., Liu, S., Lv, F., Lin, Y., Liu, G., & Peng, B. (2017) · Journal of Endodontics · doi:10.1016/j.joen.2016.12.013 · Cited by 14 · see citing works
“Autophagy is known to be involved in osteoclast differentiation and function (14, 17, 26).”
BackgroundRANKL triggers resistance to TRAIL‐induced cell death in oral squamous cell carcinoma
Ethiraj, P., Sambandam, Y., Hathaway‐Schrader, J. D., Haque, A., Novince, C. M., & Reddy, S. V. (2019) · Journal of Cellular Physiology · doi:10.1002/jcp.29086 · Cited by 14 · see citing works
“Earlier, we reported that RANKL modulates autophagic activity in OSCC cells and autophagy inhibitor suppress RANK/RANKL signaling (Sambandam et al., 2014; Sambandam, Sakamuri, Balasubramanian, & Haque, 2016).”
BackgroundSelf-citationChi, H., SON, H., Chung, Đ. C., HUAN, L., DIEM, T., & Long, L. T. (2020) · Physiological Research · doi:10.33549/physiolres.934472 · Cited by 13 · see citing works
“Increases in osteoclastogenesis has also been reported in mouse bone marrow cells under SMG conditions using autophagy markers (Sambandam et al., 2014).”
MethodMicrogravity, Bone Homeostasis, and Insulin-Like Growth Factor-1
Smith, J. K. (2020) · Applied Sciences · doi:10.3390/app10134433 · Cited by 12 · see citing works
Oligodendrocyte Progenitors Display Enhanced Proliferation and Autophagy after Space Flight
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Implications of Altered Endosome and Lysosome Biology in Space Environments
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“Differential expression of endosome proteins such as the late endosome marker Rab7 observed in spaceflight [24,25] significantly impairs the resorption capacity [4], since these proteins are integral to cell polarisation, ruffled border and osteoclast bone resorptive capability [85,86].”
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“For instance, also under the above SMG conditions, the protein expression levels of Atg5 and LC3-II increased 8.0-fold and 7.0-fold, respectively [96].”
InfluentialStrategies for studying bone loss in microgravity
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Aging and Altered Gravity: A Cellular Perspective
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“Multiple in vitro studies with either murine or human cell lines indicate increased autophagy in real and simulated microgravity [160–162].”
Chhabrani, A., Shivamallu, A. B., Bharadwaj, R. S., Kale, R., & Kathuria, A. (2024) · Acta Astronautica · doi:10.1016/j.actaastro.2024.03.014 · Cited by 4 · see citing works
Mechanisms of pyroptosis in modulating osteoblast function under simulated microgravity
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“We utilized the NASA-developed Rotary Cell Culture System (RCCS) to simulate microgravity [16–18].”
The role of autophagy during osteoclastogenesis under microgravity conditions
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“…inhibitor, to mouse bone marrow cells suppressed the expression of Atg5 and LC3, which are autophagy markers, as well as cathepsin K, which is an osteoclast marker, indicating that autophagy probably plays a role in osteoclastogenesis under microgravity conditions (Sambandam et al., 2014).”
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Bone Biology and Effects of Pharmaceutical Intervention on Bone Quality
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Autophagy development as an adaptive response to microgravity conditions in Arabidopsis thaliana
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Mimic Spaceflight: Microgravity Induces Bone Injury
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Matuszek, I., Badowska, N., Wiśniewski, B., Pachnowska, D., Pluta, N., Nowak, K. A., Rutkowska-Nowosielska, A., Czekanow, O. N., Kania-Bonicka, Z. M., & Demkow, J. (2026) · International Journal of Innovative Technologies in Social Science · doi:10.31435/ijitss.2(50).2026.5793
- 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
- PMC
- PMC4384509
- OpenAlex
- W2078536317
- Semantic Scholar
- 5e630be43646891c58ff019a97e400b353a0c892
- 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
- green
- www.ncbi.nlm.nih.gov
- 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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