TY - JOUR
T1 - A proteomics approach to study mouse long bones
T2 - examining baseline differences and mechanical loading-induced bone formation in young-adult and old mice
AU - Chermside-Scabbo, Christopher J.
AU - Shuster, John T.
AU - Erdmann-Gilmore, Petra
AU - Tycksen, Eric
AU - Zhang, Qiang
AU - Townsend, R. Reid
AU - Silva, Matthew J.
N1 - Publisher Copyright:
© 2024 Chermside-Scabbo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2024
Y1 - 2024
N2 - With aging, bone mass declines and the anabolic effects of skeletal loading diminish. While much research has focused on gene transcription, how bone ages and loses its mechanoresponsiveness at the protein level remains unclear. We developed a novel proteomics approach and performed a paired mass spectrometry and RNA-seq analysis on tibias from young-adult (5-month) and old (22-month) mice. We report the first correlation estimate between the bone proteome and transcriptome (Spearman ρ = 0.40), which is in line with other tissues but indicates that a relatively low amount of variation in protein levels is explained by the variation in transcript levels. Of 71 shared targets that differed with age, eight were associated with bone mineral density in previous GWAS, including understudied targets Asrgl1 and Timp2. We used complementary RNA in situ hybridization to confirm that Asrgl1 and Timp2 had reduced expression in osteoblasts/osteocytes in old bones. We also found evidence for reduced TGF-beta signaling with aging, in particular Tgfb2. Next, we defined proteomic changes following mechanical loading. At the protein level, bone differed more with age than with loading, and aged bone had fewer loading-induced changes. Overall, our findings underscore the need for complementary protein-level assays in skeletal biology research.
AB - With aging, bone mass declines and the anabolic effects of skeletal loading diminish. While much research has focused on gene transcription, how bone ages and loses its mechanoresponsiveness at the protein level remains unclear. We developed a novel proteomics approach and performed a paired mass spectrometry and RNA-seq analysis on tibias from young-adult (5-month) and old (22-month) mice. We report the first correlation estimate between the bone proteome and transcriptome (Spearman ρ = 0.40), which is in line with other tissues but indicates that a relatively low amount of variation in protein levels is explained by the variation in transcript levels. Of 71 shared targets that differed with age, eight were associated with bone mineral density in previous GWAS, including understudied targets Asrgl1 and Timp2. We used complementary RNA in situ hybridization to confirm that Asrgl1 and Timp2 had reduced expression in osteoblasts/osteocytes in old bones. We also found evidence for reduced TGF-beta signaling with aging, in particular Tgfb2. Next, we defined proteomic changes following mechanical loading. At the protein level, bone differed more with age than with loading, and aged bone had fewer loading-induced changes. Overall, our findings underscore the need for complementary protein-level assays in skeletal biology research.
KW - RNA-seq/transcriptomics
KW - aging
KW - bone
KW - mechanical loading
KW - proteomics
UR - http://www.scopus.com/inward/record.url?scp=85206471890&partnerID=8YFLogxK
U2 - 10.18632/aging.206131
DO - 10.18632/aging.206131
M3 - Article
C2 - 39400554
AN - SCOPUS:85206471890
SN - 1945-4589
VL - 16
SP - 12726
EP - 12768
JO - Aging
JF - Aging
IS - 19
ER -