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Overexpression of spermidine/spermine N 1-acetyltransferase impairs osteoblastogenesis and alters mouse bone phenotype

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Abstract

Spermidine/spermine N 1-acetyltransferase (SSAT) is a catabolic regulator of polyamines, ubiquitous molecules essential for cell proliferation and differentiation. In pathological conditions, the increased polyamine catabolism has been shown to mediate its cellular functions not only by changed polyamine levels but also by the availability of metabolites shared with other metabolic pathways or by production of toxic compounds. Our previous results showed that mice overexpressing SSAT (SSAT mice) developed a myeloproliferative disease and the bone marrow microenvironment partly contributed to its development. In this study, the physiological role of SSAT and polyamines in bone remodeling was characterized. Skeletal development of the SSAT mice appeared outwardly similar to wild-type mice until maturity, after which the SSAT mice developed kyphosis. With aging, the SSAT overexpression elicited increased bone perimeter with strikingly thinned cortical bone, decreased trabecular thickness and increased trabecular number in mice. In vitro studies showed that the maturation of SSAT overexpressing osteoblasts was impaired and the expression of bone formation marker genes was dramatically decreased. The polyamine pattern in osteoblasts of SSAT mice was distorted in comparison with wild-type mice. However, treatment of osteoblasts with a SSAT-inducing functional polyamine analogue suggested that defective osteoblastogenesis resulted rather from other consequences of enhanced SSAT activity than lowered levels of the higher polyamines. In comparison to SSAT overexpressing mice, SSAT deficiency led to opposite changes in osteoblastogenesis and differences in bone phenotype in mice. In conclusion, the level of SSAT enzyme activity affected osteoblastogenesis and hence influenced bone remodeling and the bone phenotype in mice. Furthermore, our results suggest the contribution of the catabolic part of the polyamine cycle, other than polyamine depletion, in pathophysiological processes of bone remodeling.

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Abbreviations

αMetSPD:

α-Methylspermidine

ALP:

Alkaline phosphatase

APAO:

Acetylpolyamine oxidase

BMD:

Bone mineral density

B.Pm:

Bone perimeter

BSP:

Bone sialoprotein

BV:

Bone volume

Cs.Th:

Cortical thickness

Ct.Ar:

Cortical area

CTSK:

Cathepsin K

CTX:

Carboxy-terminal cross-linking telopeptide

MSC:

Mesenchymal stromal cell

NFATC1:

Nuclear factor of activated T-cells cytoplasmic 1

OC:

Osteocalcin

ODC:

Ornithine decarboxylase

OPN:

Osteopontin

OSX:

Osterix

PINP:

Procollagen type I propeptide

Po.V:

Total volume of porosity

ROS:

Reactive oxygen species

RUNX2:

Runt-related transcription factor 2

SMI:

Structural model index

SSAT:

Spermidine/spermine N 1-acetyltransferase

SSATKO:

Spermidine/spermine N 1-acetyltransferase knockout

Tb.Th:

Trabecular thickness

Tb.N:

Trabecular number

Tb.Sp:

Trabecular separation

TRAcP:

Tartrate-resistant acid phosphatase

Tt.Ar:

Total periosteal area

TV:

Tissue volume

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Acknowledgments

We gratefully acknowledge Ms Tuula Reponen, Ms Anne Karppinen, Ms Arja Korhonen and Ms Marita Heikkinen for their skilful technical assistance. We thank Professor Alex Khomutov for providing alpha-methylated spermidine analogue.

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Authors and Affiliations

  1. Biotechnology and Molecular Medicine, A.I. Virtanen Institute for Molecular Sciences, Biocenter Kuopio, University of Eastern Finland, P.O. Box 1627, 70211, Kuopio, Finland

    Sini Pirnes-Karhu, Leena Alhonen & Anne Uimari

  2. School of Pharmacy, University of Eastern Finland, 70211, Kuopio, Finland

    Jorma Määttä

  3. Department of Cell Biology and Anatomy, University of Turku, 20014, Turku, Finland

    Jorma Määttä

  4. Department of Medical Technology, University of Oulu, 90014, Oulu, Finland

    Mikko Finnilä

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  1. Sini Pirnes-Karhu
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  2. Jorma Määttä
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  4. Leena Alhonen
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  5. Anne Uimari
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Correspondence to Sini Pirnes-Karhu.

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Pirnes-Karhu, S., Määttä, J., Finnilä, M. et al. Overexpression of spermidine/spermine N 1-acetyltransferase impairs osteoblastogenesis and alters mouse bone phenotype. Transgenic Res 24, 253–265 (2015). https://doi.org/10.1007/s11248-014-9836-6

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  • DOI: https://doi.org/10.1007/s11248-014-9836-6

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