Dephosphorylated uncarboxylated Matrix-Gla-Protein as candidate biomarker for immune-mediated vascular remodeling and prognosis in pulmonary hypertension

Figures

• 

  Figure 1. Plasma levels of dp-ucMGP in IPAH, CTD-PAH, CTEPH, and CTD patients. (A) Plasma levels of dp-ucMGP in IPAH (n = 26), CTD-PAH (n = 8), CTEPH (n = 11) and CTD (n = 28) patients. (B) Plasma levels of dp-ucMGP in non-IMID-PAH (n = 8) and IMID-PAH (n = 26) patients. Data are shown as symbols for individual patients or HCs. Patients depicted as red dots are categorized as CTD-PAH (n = 8), black dots depict IPAH patients (n = 18). The horizontal dotted line represents the mean value of dp-ucMGP in aged-matched healthy controls (393pmol/L). Horizontal gray bars represent median values with interquartile range. Statistical analysis was performed using the Kruskal-Wallis and Mann-Whitney test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. dp-ucMGP, dephosphorylated-uncarboxylated Matrix-GLA-Protein; IPAH, idiopathic pulmonary hypertension; CTD-PAH, connective tissue associated pulmonary arterial hypertension; CTEPH, chronic thrombo-embolic pulmonary hypertension; CTD, connective tissue disease; IMID, immune mediated inflammatory disease.

• 

  Figure 2. Amount of plasma-induced calcification of induced vascular smooth muscle cells. (A) Plasma-induced iVSMC calcification in IPAH (n = 26), CTD-PAH (n = 8), CTEPH (n = 11), and CTD (n = 28) patients. (B) plasma-induced iVSMC calcification in IPAH non-IMID-PAH (n = 8) and IMID-PAH (n = 26) patients. Data are shown as symbols for individual patients or HCs. Horizontal bars represent median values with interquartile range. Patients depicted as red dots are categorized as CTD-PAH (n = 8), black dots depict IPAH patients (n = 18). Statistical analysis was performed using the Kruskal-Wallis and Mann-Whitney test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. RFP, red fluorescent protein; HC, healthy controls; IPAH, idiopathic pulmonary hypertension; CTD-PAH, connective tissue associated pulmonary arterial hypertension; CTEPH, chronic thrombo-embolic pulmonary hypertension; CTD, connective tissue disease; IMID, immune-mediated inflammatory disease.

• 

  Figure 3. Plasma levels of different soluble immune checkpoint proteins (sICPs) in IPAH, CTD patients and healthy controls. The indicated sICPs were measured in plasma samples of 24 IPAH patients and 27 CTD patients. sICPs are classified as either co-stimulatory or co-inhibitory and are found on either T-cells or antigen-presenting cells. Horizontal bars represent median values with interquartile range. Statistical analysis was performed using the Kruskal-Wallis test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. sPD1, programmed death protein-1; sPDL2, programmed death ligand-2; sBTLA, soluble B- and T-lymphocyte attenuator; cluster of differentiation (CD)27, CD28, CD80, CD137 and CD152 (CTLA4/cytotoxic T-lymphocyte-associated antigen-3; sGITR, glucocorticoid-induced TNFR-related protein; sHVEM herpesvirus entry mediator; sIDO, indoleamine 2,3-dioxygenase; sLAG3, Lymphocyte-activation gene 3; sTIM3, T-cell immunoglobulin and mucin-domain containing-3.

• 

  Figure 4. Correlation matrices for different soluble immune checkpoint proteins (sICPs) with dp-ucMGP and calcification in IPAH (n = 24) and CTD (n = 27) patients. Correlation coefficient was calculated using nonparametric Spearman pairwise correlation. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

• 

  Figure 5. Plasma levels of different soluble immune checkpoint proteins (sICPs) in non-IMID-PAH and IMID-PAH patients. The indicated sICPs were measured in plasma samples of non-IMID-PAH (n = 8) patients and IMID-PAH (n = 19) patients. Horizontal bars represent median values with interquartile range. Statistical analysis was performed using the Mann-Whitney test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

• 

  Figure 6. Kaplan-Meyer survival analyses for dp-ucMGP high/low of IPAH patients (n = 26). High/low cut-off values were based on the 50th percentile of dp-ucMGP plasma levels. Statistical analysis was performed using the log-rank test.