Oral Presentation ANZBMS-MEPSA-ANZORS 2022

The effects of GCSF in human and murine osteoclast differentiation and mature function models (#214)

Rouha MS Granfar 1 , Nigel A Morrison 1
  1. Griffith University, Southport, QUEENSLAND, Australia

CSF3 or granulocyte colony stimulating factor (GCSF) has clinically documented effects on bone homeostasis in health, pathology, and accompanying therapeutic applications by either directly or indirectly acting on mature osteoclasts or precursors. GCSF appears to stimulate osteoclasts (OC), impair osteoblasts (OB), elevated in tissue and serology of periodontitis, mobilise haematopoetic stem cells (HSC) and alter tumour burden in particular by disrupting the CXCR4:CXCL12 axis [8,9,11,12,13,14,15,16,18]. 

Our research group detected 11.8-fold up-regulation of GCSF receptor in human osteoclasts derived from adherent Peripheral Blood Mononuclear Cells (PBMC) [1] and indicated increased expression and involvement of C-C chemokine receptors and ligands (CCR2/CCL2 & CCL5) within OC and monocyte derived multinuclear cell fusion [1,2,3,4,5]. The involvement of GCSF in modulating CCL2 in neutrophil differentiation, neuropathic pain pathways, and Craniocervical Instability (CCI) supports further investigation of GCSF in musculoskeletal biology, pathology, and the potential as a broader therapeutic target [9,10,17].

Initial results of GCSF-treatment in models of osteoclast differentiation and mature OC function will be presented, including gene expression, dentine resorption, and cell morphology profiles of osteoclast models derived from human peripheral blood and mouse bone marrow monocytes (wild type, CCL2 -/-; CCR2 -/-) with either "continuous", "early" (0h to pre-osteoclast or fused polykaryons), and "late" (polykaryon to activated/mature) treatment [1,2,3,4,5,6,7]. 

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  2. Day CJ, Kim MS, Lopez CM, Nicholson GC, & Morrison NA. (2005). NFAT expression in human osteoclasts. Journal of Cellular Biochemistry, 95(1), 17–23. https://doi.org/10.1002/jcb.20410
  3. Granfar RMS, Day CJ, Kim MS, & Morrison NA. (2005). Optimised real-time quantitative PCR assays for RANKL regulated genes. Molecular and Cellular Probes, 19(2), 119–126. https://doi.org/10.1016/j.mcp.2004.10.003
  4. Khan UA, Hashimi SM, Bakr MM, Forwood MR, & Morrison NA. (2016). CCL2 and CCR2 are Essential for the Formation of Osteoclasts and Foreign Body Giant Cells. Journal of Cellular Biochemistry, 117(2), 382–389. https://doi.org/10.1002/jcb.25282
  5. Kim MS, Magno CL, Day CJ, & Morrison NA. (2006). Induction of chemokines and chemokine receptors CCR2b and CCR4 in authentic human osteoclasts differentiated with RANKL and osteoclast like cells differentiated by MCP-1 and RANTES. Journal of Cellular Biochemistry, 97(3), 512–518. https://doi.org/10.1002/jcb.20649
  6. Selinger CI, Day CJ, & Morrison NA. (2005). Optimized transfection of diced siRNA into mature primary human osteoclasts: Inhibition of cathepsin K mediated bone resorption by siRNA. Journal of Cellular Biochemistry, 96(5), 996–1002. https://doi.org/10.1002/jcb.20575
  7. Brazier H, Stephens S, Ory S, Fort P, Morrison N, & Blangy A. (2006). Expression Profile of RhoGTPases and RhoGEFs During RANKL-Stimulated Osteoclastogenesis: Identification of Essential Genes in Osteoclasts. Journal of Bone and Mineral Research, 21(9), 1387–1398. https://doi.org/10.1359/jbmr.060613
  8. Yu H, Zhang T, Lu H, Ma Q, Zhao D, Sun J & Wang Z. (2021). Granulocyte colony-stimulating factor (G-CSF) mediates bone resorption in periodontitis. BMC Oral Health 21, 299. https://doi.org/10.1186/s12903-021-01658-1
  9. Iida S, Kohro T, Kodama T, Nagata S, Fukunaga R. Identification of CCR2, flotillin, and gp49B genes as new G-CSF targets during neutrophilic differentiation. J Leukoc Biol. (2005) Aug;78(2):481-90. doi: 10.1189/jlb.0904515. Epub 2005 May 13. PMID: 15894583.
  10. McRae JL, Vikstrom IB, Bongoni AK, Salvaris EJ, Fisicaro N, Ng M, Alhamdoosh M, Baz Morelli A, Cowan PJ, Pearse MJ. (2020). Blockade of the G-CSF Receptor Is Protective in a Mouse Model of Renal Ischemia-Reperfusion Injury. J Immunol. Sep 1;205(5):1433-1440. doi: 10.4049/jimmunol.2000390. Epub 2020 Jul 27. PMID: 32839213.
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  13. Takamatsu Y, Simmons PJ, Moore RJ, Morris HA, To LB, & Lévesque JP. (1998). Osteoclast-mediated bone resorption is stimulated during short-term administration of granulocyte colony-stimulating factor but is not responsible for hematopoietic progenitor cell mobilization. Blood, 92(9), 3465–3473.
  14. Soshi S, Takahashi HE, Tanizawa T, Endo N, Fujimoto R, & Murota, K. (1996). Effect of recombinant human granulocyte colony-stimulating factor (rh G-CSF) on rat bone: Inhibition of bone formation at the endosteal surface of vertebra and tibia. Calcified Tissue International, 58(5), 337–340. https://doi.org/10.1007/BF02509382
  15. Lévesque J-P, Hendy J, Takamatsu Y, Simmons PJ, & Bendall LJ. (2003). Disruption of the CXCR4/CXCL12 chemotactic interaction during hematopoietic stem cell mobilization induced by GCSF or cyclophosphamide. The Journal of Clinical Investigation, 111(2), 187–196. https://doi.org/10.1172/JCI15994
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