Differentially expressed circulating microRNAs in the development of acute diabetic Charcot foot

Jennifer Pasquier, Vimal Ramachandran, Moh'd Rasheed Abu-Qaoud, Binitha Thomas, Manasi J Benurwar, Omar Chidiac, Jessica Hoarau-Véchot, Amal Robay, Khalid Fakhro, Robert A Menzies, Amin Jayyousi, Mahmoud Zirie, Jassim Al Suwaidi, Rayaz A Malik, Talal K Talal, Seyed Hani Najafi-Shoushtari, Arash Rafii, Charbel Abi Khalil, Jennifer Pasquier, Vimal Ramachandran, Moh'd Rasheed Abu-Qaoud, Binitha Thomas, Manasi J Benurwar, Omar Chidiac, Jessica Hoarau-Véchot, Amal Robay, Khalid Fakhro, Robert A Menzies, Amin Jayyousi, Mahmoud Zirie, Jassim Al Suwaidi, Rayaz A Malik, Talal K Talal, Seyed Hani Najafi-Shoushtari, Arash Rafii, Charbel Abi Khalil

Abstract

Aim: Charcot foot (CF) is a rare complication of Type 2 diabetes (T2D).

Materials & methods: We assessed circulating miRNAs in 17 patients with T2D and acute CF (G1), 17 patients with T2D (G2) and equivalent neuropathy and 17 patients with T2D without neuropathy (G3) using the high-throughput miRNA expression profiling.

Results: 51 significantly deregulated miRNAs were identified in G1 versus G2, 37 in G1 versus G3 and 64 in G2 versus G3. Furthermore, we demonstrated that 16 miRNAs differentially expressed between G1 versus G2 could be involved in osteoclastic differentiation. Among them, eight are key factors involved in CF pathophysiology.

Conclusion: Our data reveal that CF patients exhibit an altered expression profile of circulating miRNAs.

Trial registration: ClinicalTrials.gov NCT02316483.

Keywords: Charcot foot; diabetes; epigenetics; inflammation; micro-RNA.

Conflict of interest statement

Financial & competing interests disclosure

This work was made possible by a national research priority program (NPRP) grant 7-701-3-192 from the Qatar National Research Fund (a member of Qatar Foundation). The findings achieved herein are solely the responsibility of the authors. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Figures

Figure 1. . Volcano plots.
Figure 1.. Volcano plots.
Volcano plots showing differentially expressed miRNAs in the following comparisons: T2D with Charcot foot over T2D with neuropathy (Group 1 vs Group 2) (A); T2D with Charcot foot over T2D without neuropathy (Group 1 vs Group 3) (B); and T2D with neuropathy over T2D without neuropathy (Group 2 vs Group 3) (C). miRNAs with more than twofold change and a significant p-value after applying Bonferroni correction are shown in green. Those, which show more than twofold change but whose p-value significance is achieved by discounting Bonferroni correction are shown in yellow. Gray dots represent miRNAs exhibiting less than twofold change, while red dots stand for those showing more than twofold change but lacking statistical significance. Solid vertical lines indicate twofold upregulation or downregulation on the log scale. Dashed horizontal lines either mark p-value = 0.05 on the log scale (red line), or Bonferroni-corrected p-value cut-off (green line). T2D: Type 2 diabetes.
Figure 2. . Venn diagram.
Figure 2.. Venn diagram.
A three-way Venn diagram representing the apparition of miRNAs in each comparison. The one in the center (black) appears in every contrast, the one at the merging of two circles (orange, purple, green) appears in two comparisons and the one in one circle only (yellow, red, blue) appears in only one set of data.
Figure 3. . A network of predicted…
Figure 3.. A network of predicted genes affected by differentially expressed miRNA from G1 versus G2 comparison created by Ingenuity Pathway Analysis.
The networks hypothesized by Ingenuity Pathway Analysis are based on the molecular relationships, interactions and pathway associations between the methylated candidate genes as shown in a graphical representation.
Figure 4. . Role of miRNA differentially…
Figure 4.. Role of miRNA differentially expressed between G1 versus G2 in the osteoclast differentiation.
The graphical representation had been generated by Ingenuity Pathway Analysis and reflects the molecular relationships between miRNAs and candidate genes. miRNAs in red were overexpressed in Charcot foot (CF) patients as compared with Type 2 diabetes (T2D) patients with neuropathy, while the ones in green were downexpressed in CF patients compared with T2D patients with neuropathy.

References

    1. Abi Khalil C, Roussel R, Mohammedi K, Danchin N, Marre M. Cause-specific mortality in diabetes: recent changes in trend mortality. Eur. J. Prev. Cardiol. 2012;19(3):374–381.
    1. Huang D, Refaat M, Mohammedi K, Jayyousi A, Al Suwaidi J, Abi Khalil C. Macrovascular complications in patients with diabetes and prediabetes. Biomed. Res. Int. 2017;2017:7839101.

    2. • A review that summarizes well cardiovascular complications of diabetes.

    1. Pop-Busui R, Boulton AJ, Feldman EL, et al. Diabetic neuropathy: a position statement by the American Diabetes Association. Diabetes Care. 2017;40(1):136–154.
    1. Nobrega MB, Aras R, Netto EM, et al. Risk factors for Charcot foot. Arch. Endocrinol. Metab. 2015;59(3):226–230.
    1. Tuttolomondo A, Maida C, Pinto A. Diabetic foot syndrome as a possible cardiovascular marker in diabetic patients. J. Diabetes Res. 2015;2015:268390.

    2. • Links diabetic foot syndrome with cardiovascular risk.

    1. Young MJ, Marshall A, Adams JE, Selby PL, Boulton AJ. Osteopenia, neurological dysfunction, and the development of Charcot neuroarthropathy. Diabetes Care. 1995;18(1):34–38.
    1. Tuttolomondo A, Maida C, Pinto A. Diabetic foot syndrome: Immune-inflammatory features as possible cardiovascular markers in diabetes. World J. Orthop. 2015;6(1):62–76.
    1. Mabilleau G, Petrova N, Edmonds ME, Sabokbar A. Number of circulating CD14-positive cells and the serum levels of TNF-α are raised in acute Charcot foot. Diabetes Care. 2011;34(3):e33.
    1. Pasquier J, Thomas B, Hoarau-Vechot J, et al. Circulating microparticles in acute diabetic Charcot foot exhibit a high content of inflammatory cytokines, and support monocyte-to-osteoclast cell induction. Sci. Rep. 2017;7(1):16450.

    2. •• Describes the role of microparticles in Charcot foot (CF) disease.

    1. Petrova NL, Edmonds ME. Acute Charcot neuro-osteoarthropathy. Diabetes Metab. Res. Rev. 2016;32(Suppl. 1):281–286.

    2. •• Up-to-date review about CF disease.

    1. Uccioli L, Sinistro A, Almerighi C, et al. Proinflammatory modulation of the surface and cytokine phenotype of monocytes in patients with acute Charcot foot. Diabetes Care. 2010;33(2):350–355.
    1. Mabilleau G, Petrova NL, Edmonds ME, Sabokbar A. Increased osteoclastic activity in acute Charcot's osteoarthropathy: the role of receptor activator of nuclear factor-κB ligand. Diabetologia. 2008;51(6):1035–1040.
    1. Bartel DP. miRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116(2):281–297.

    2. •• An important review regarding miRNAs.

    1. Arroyo JD, Chevillet JR, Kroh EM, et al. Argonaute2 complexes carry a population of circulating miRNAs independent of vesicles in human plasma. Proc. Natl Acad. Sci. USA. 2011;108(12):5003–5008.
    1. Chen X, Ba Y, Ma L, et al. Characterization of miRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res. 2008;18(10):997–1006.
    1. Weber JA, Baxter DH, Zhang S, et al. The miRNA spectrum in 12 body fluids. Clin. Chem. 2010;56(11):1733–1741.
    1. Da Silveira JC, Veeramachaneni DN, Winger QA, Carnevale EM, Bouma GJ. Cell-secreted vesicles in equine ovarian follicular fluid contain miRNAs and proteins: a possible new form of cell communication within the ovarian follicle. Biol. Reprod. 2012;86(3):71.
    1. Thomas MR, Lip GY. Novel risk markers and risk assessments for cardiovascular disease. Circ. Res. 2017;120(1):133–149.
    1. Bayraktar R, Van Roosbroeck K, Calin GA. Cell-to-cell communication: miRNAs as hormones. Mol. Oncol. 2017;11(12):1673–1686.
    1. Borujeni MJS, Esfandiary E, Taheripak G, Codoner-Franch P, Alonso-Iglesias E, Mirzaei H. Molecular aspects of diabetes mellitus: resistin, miRNA and exosome. J. Cell Biochem. 2018;119(2):1257–1272.
    1. Pasquier J, Hoarau-Vechot J, Fakhro K, Rafii A, Abi Khalil C. Epigenetics and cardiovascular disease in diabetes. Curr. Diab. Rep. 2015;15(12):108.

    2. •• Comprehensive review regarding epigenetics of diabetes complications

    1. Rogers LC, Frykberg RG, Armstrong DG, et al. The Charcot foot in diabetes. Diabetes Care. 2011;34(9):2123–2129.
    1. Sun Y, Peng R, Peng H, et al. miR-451 suppresses the NF-κB-mediated proinflammatory molecules expression through inhibiting LMP7 in diabetic nephropathy. Mol. Cell Endocrinol. 2016;433:75–86.
    1. Kocijan R, Muschitz C, Geiger E, et al. Circulating miRNA signatures in patients with idiopathic and postmenopausal osteoporosis and fragility fractures. J. Clin. Endocrinol. Metab. 2016;101(11):4125–4134.
    1. Kim BS, Jung JY, Jeon JY, Kim HA, Suh CH. Circulating hsa-miR-30e-5p, hsa-miR-92a-3p, and hsa-miR-223-3p may be novel biomarkers in systemic lupus erythematosus. HLA. 2016;88(4):187–193.
    1. Madhavan D, Peng C, Wallwiener M, et al. Circulating miRNAs with prognostic value in metastatic breast cancer and for early detection of metastasis. Carcinogenesis. 2016;37(5):461–470.
    1. Khan FH, Pandian V, Ramraj S, Aravindan S, Herman TS, Aravindan N. Reorganization of metastamiRs in the evolution of metastatic aggressive neuroblastoma cells. BMC Genomics. 2015;16:501.
    1. Yan H, Wang S, Yu H, Zhu J, Chen C. Molecular pathways and functional analysis of miRNA expression associated with paclitaxel-induced apoptosis in hepatocellular carcinoma cells. Pharmacology. 2013;92(3–4):167–174.
    1. Zou M, Wang F, Gao R, et al. Autophagy inhibition of hsa-miR-19a-3p/19b-3p by targeting TGF-β R II during TGF-β1-induced fibrogenesis in human cardiac fibroblasts. Sci. Rep. 2016;6:24747.
    1. Tang Y, Zhang YC, Chen Y, Xiang Y, Shen CX, Li YG. The role of miR-19b in the inhibition of endothelial cell apoptosis and its relationship with coronary artery disease. Sci. Rep. 2015;5:15132.
    1. Calimlioglu B, Karagoz K, Sevimoglu T, Kilic E, Gov E, Arga KY. Tissue-specific molecular biomarker signatures of Type 2 diabetes: an integrative analysis of transcriptomics and protein–protein interaction data. OMICS. 2015;19(9):563–573.
    1. Liu D, Li YW, Luo G, et al. LncRNA SPRY4-IT1 sponges miR-101-3p to promote proliferation and metastasis of bladder cancer cells through up-regulating EZH2*. Cancer Lett. 2017;388:281–291.
    1. Chang L, Yuan YF, Li CC, et al. Upregulation of SNHG6 regulates ZEB1 expression by competitively binding miR-101-3p and interacting with UPF1 in hepatocellular carcinoma. Cancer Lett. 2016;383(2):183–194.
    1. Sun YX, Zhang JF, Xu J, et al. miRNA-144-3p inhibits bone formation in distraction osteogenesis through targeting Connexin 43. Oncotarget. 2017;8(52):89913–89922.
    1. Song J, Kang YH, Yoon S, Chun CH, Jin EJ. HIF-1a: CRAT: miR-144-3p axis dysregulation promotes osteoarthritis chondrocyte apoptosis and VLCFA accumulation. Oncotarget. 2017;8(41):69351–69361.
    1. Wang Y, Zhang YY, Yang T, et al. Long non-coding RNA MALAT1 for promoting metastasis and proliferation by acting as a ceRNA of miR-144-3p in osteosarcoma cells. Oncotarget. 2017;8(35):59417–59434.
    1. Shen HX, Li WJ, Tian Y, et al. Upregulation of miR-362-3p Modulates proliferation and anchorage-independent growth by directly targeting Tob2 in hepatocellular carcinoma. J. Cell. Biochem. 2015;116(8):1563–1573.
    1. Christensen LL, Tobiasen H, Holm A, et al. miRNA-362-3p induces cell cycle arrest through targeting of E2F1, USF2 and PTPN1 and is associated with recurrence of colorectal cancer. Int. J. Cancer. 2013;133(1):67–78.
    1. Wang K, Yuan Y, Cho JH, McClarty S, Baxter D, Galas DJ. Comparing the miRNA spectrum between serum and plasma. PLoS ONE. 2012;7(7):e41561.
    1. Wang K, Zhang S, Weber J, Baxter D, Galas DJ. Export of miRNAs and miRNA-protective protein by mammalian cells. Nucliec Acids Res. 2010;38(20):7248–7259.
    1. Mestdagh P, Hartmann N, Baeriswyl L, et al. Evaluation of quantitative miRNA expression platforms in the miRNA quality control (miRQC) study. Nat. Methods. 2014;11(8):809–815.
    1. Li Q, Chen L, Chen D, Wu X, Chen M. Influence of miRNA-related polymorphisms on clinical outcomes in coronary artery disease. Am. J. Transl. Res. 2015;7(2):393–400.
    1. Neal CS, Michael MZ, Pimlott LK, Yong TY, Li JY, Gleadle JM. Circulating miRNA expression is reduced in chronic kidney disease. Nephrol. Dial. Transplant. 2011;26(11):3794–3802.

Source: PubMed

Sponsors and Collaborators

Medical Conditions

Drug Interventions

3
Subscribe