Expression of immune-related genes in peripheral blood leukocytes of postmenopausal osteoporosis patients

doi:10.3969/j.issn.2095-4344.2090

Abstract

Abstract:

BACKGROUND: Bones are currently considered as an immune organ. A variety of immune cells that originate from the bone marrow can interact with the cells of the skeletal system to jointly regulate bone metabolism. Explorations on the pathogenesis of postmenopausal osteoporosis as well as treatment-related molecular targets and signal pathways can help prevention and treatment of the disease.

OBJECTIVE: To investigate the expression profiles of immune-related genes in peripheral blood leukocytes of postmenopausal osteoporosis patients using RNA-Seq technology.

METHODS: Forty female patients who had experienced menopause for 0 to 20 years and were hospitalized due to fractures were enrolled. They were divided into normal bone mass group (T > -1) and osteoporosis group (T < -2.5) by scanning with Lunar Prodigy dual-energy X-ray bone densitometer. Then, the total RNA of leukocytes in the peripheral blood samples from five patients of each group was extracted. Differentially expressed genes between two groups were detected by RNA-Seq technology followed by GO enrichment analysis. The immune-related genes were screened and analyzed by KEGG signal pathway. Peripheral blood samples from 20 patients of each group were collected and the results of KEGG PATHWAY bioinformatics analysis were verified by real-time PCR. The implementation of the study plan complied with the relevant ethical requirements of the Pudong New Area Gongli Hospital, Shanghai (approval No. 20170301).

RESULTS AND CONCLUSION: Compared with the normal bone mass group, 187 genes were significantly changed in the osteoporosis group (fold change > 2), and 131 genes were up-regulated and 56 genes were down-regulated. We identified in total 29 differentially expressed immune-related genes including 25 up-regulated and 4 down-regulated ones. There was significant difference in expression between the osteoporosis and normal bone mass groups for genes, including KIR3DL1, KIR3DL2, KIR2DL4, KLRD1 and HSPA6 (P < 0.05). These differentially expressed genes are potentially important for the natural killer cell-mediated cytotoxicity by the KEGG pathway analysis. KIR3DL1, KIR3DL2, KIR2DL4, KLRD1 and HSPA6 may be closely related to the natural killer cell-mediated cytotoxicity during the occurrence of postmenopausal osteoporosis.

Key words:

postmenopausal osteoporosis, peripheral blood leukocytes, RNA sequence, immune-related genes

CLC Number:

Chen Tianning, Yang Tieyi, Shao Jin, Kong Dece, Liu Shuyi, Zhou Wenchao, Zhang Yan.

Expression of immune-related genes in peripheral blood leukocytes of postmenopausal osteoporosis patients [J]. Chinese Journal of Tissue Engineering Research, 2020, 24(25): 4033-4038.

Figures/Tables 5

References

[1] COSMAN F.Long-term treatment strategies for postmenopausal osteoporosis. Curr Opin Rheumatol. 2018; 30(4):420-426.

[2] WATTS NB. Postmenopausal Osteoporosis: A Clinical Review. J Womens Health (Larchmt). 2018;27(9):1093-1096.

[3] ARRON JR, CHOI Y. Bone versus immune system. Nature. 2000;408(6812):535-536.

[4] TAKAYANAGI H, OGASAWARA K, HIDA S, et al. T-cell-mediated regulation of osteoclastogenesis by signalling cross-talk between RANKL and IFN-gamma. Nature. 2000; 408(6812):600-605.

[5] MORI G,D'AMELIO P,FACCIO R, et al. Bone-immune cell crosstalk: bone diseases.J Immunol Res. 2015; 2015:108451.

[6] ANDERS S, HUBER W. Differential expression of RNA-Seq data at the gene level–the DESeq package. Heidelberg, Germany: European Molecular Biology Laboratory (EMBL). 2012.

[7] ZVYAGIN IV, MAMEDOV IZ, BRITANOVA OV, et al. Contribution of functional KIR3DL1 to ankylosing spondylitis. Cell Mol Immunol. 2010;7(6):471-476.

[8] BATTISTELLA M, JANIN A, JEAN-LOUIS F, et al. KIR3DL2 (CD158k) is a potential therapeutic target in primary cutaneous anaplastic large-cell lymphoma. Br J Dermatol. 2016;175(2):325-333.

[9] NOWAK I,MAJORCZYK E,PŁOSKI R,et al.Lack of KIR2DL4 gene in a fertile Caucasian woman.Tissue Antigens. 2011; 78(2):115-119.

[10] SHIN DL, PANDEY AK, ZIEBARTH JD, et al. Segregation of a spontaneous Klrd1 (CD94) mutation in DBA/2 mouse substrains. G3 (Bethesda). 2014;5(2):235-239.

[11] REGELING A, IMHANN F, VOLDERS HH, et al. HSPA6 is an ulcerative colitis susceptibility factor that is induced by cigarette smoke and protects intestinal epithelial cells by stabilizing anti-apoptotic Bcl-XL. Biochim Biophys Acta. 2016; 1862(4):788-796.

[12] NILSSON S, KOEHLER KF, GUSTAFSSON JÅ. Development of subtype-selective oestrogen receptor-based therapeutics. Nat Rev Drug Discov. 2011;10(10):778-792.

[13] MIYAUCHI Y, SATO Y, KOBAYASHI T, et al. HIF1α is required for osteoclast activation by estrogen deficiency in postmenopausal osteoporosis. Proc Natl Acad Sci U S A. 2013;110(41):16568-16573.

[14] STREICHER C, HEYNY A, ANDRUKHOVA O, et al. Estrogen Regulates Bone Turnover by Targeting RANKL Expression in Bone Lining Cells. Sci Rep. 2017;7(1):6460.

[15] RIZZOLI R.Postmenopausal osteoporosis: Assessment and management. Best Pract Res Clin Endocrinol Metab. 2018; 32(5):739-757.

[16] SRIVASTAVA RK, DAR HY, MISHRA PK. Immunoporosis: Immunology of Osteoporosis-Role of T Cells. Front Immunol. 2018;9:657.

[17] TILKERIDIS K, KIZIRIDIS G, VERVERIDIS A, et al. Immunoporosis: A New Role for Invariant Natural Killer T (NKT) Cells Through Overexpression of Nuclear Factor-kappaB Ligand (RANKL). Med Sci Monit. 2019;25: 2151-2158.

[18] LISOVSKY I, KANT S, TREMBLAY-MCLEAN A, et al. Differential contribution of education through KIR2DL1, KIR2DL3, and KIR3DL1 to antibody-dependent (AD) NK cell activation and ADCC. J Leukoc Biol. 2019;105(3):551-563.

[19] CHENG H, SCHWELL V, CURTIS BR, et al. Conformational Changes in the Cytoplasmic Region of KIR3DL1 upon Interaction with SHP-2. Structure. 2019;27(4):639-650 e2.

[20] URESHINO H, SHINDO T, KOJIMA H, et al. Allelic Polymorphisms of KIRs and HLAs Predict Favorable Responses to Tyrosine Kinase Inhibitors in CML. Cancer Immunol Res. 2018;6(6):745-754.

[21] KAMIYA T, SEOW SV, WONG D, et al. Blocking expression of inhibitory receptor NKG2A overcomes tumor resistance to NK cells. J Clin Invest. 2019;129(5):2094-2106.

[22] KORDELAS L, STECKEL NK, HORN PA, et al. The Activating NKG2C Receptor Is Significantly Reduced in NK Cells after Allogeneic Stem Cell Transplantation in Patients with Severe Graft-versus-Host Disease.Int J Mol Sci. 2016 Oct 27;17(11). pii: E1797.

[23] HUANG H, PATEL DD, MANTON KG. The immune system in aging: roles of cytokines, T cells and NK cells. Front Biosci. 2005;10: 192-215.

[24] LI L, WANG Y, ZHANG N,et al. Heterozygous deletion of LRP5 gene in mice alters profile of immune cells and modulates differentiation of osteoblasts. Bioscience trends. 2018;12(3): 266-274.

[25] SCHILLING D, TETZLAFF F, KONRAD S, et al. A hypoxia-induced decrease of either MICA/B or Hsp70 on the membrane of tumor cells mediates immune escape from NK cells. Cell Stress Chaperones. 2015;20(1):139-147.