Reduced levels of CCL2 and CXCL10 in systemic lupus erythematosus patients under treatment with prednisone, mycophenolate mofetil, or hydroxychloroquine, except in a high STAT1 subset - PubMed (original) (raw)

Reduced levels of CCL2 and CXCL10 in systemic lupus erythematosus patients under treatment with prednisone, mycophenolate mofetil, or hydroxychloroquine, except in a high STAT1 subset

Paul R Dominguez-Gutierrez et al. Arthritis Res Ther. 2014.

Abstract

Introduction: Our recent data showed that signal transducers and activators of transcription 1 (STAT1), adenosine deaminase acting on RNA (ADAR), C-C motif chemokine ligand 2 (CCL2), and C-X-C motif chemokine 10 (CXCL10) were significantly elevated in a systemic lupus erythematosus (SLE) cohort compared to healthy donors. High and low STAT1 subsets were identified in SLE patient visits. The present study analyzed the correlation of common treatments used in SLE with the levels of these biomarkers.

Methods: Peripheral blood leukocytes were collected from 65 healthy donors and 103 SLE patients, of whom 60 had samples from two or more visits. Total RNA was isolated and analyzed for the expression of mRNA and microRNA using Taqman real-time polymerase chain reaction (PCR) assays. Relative expression of interferon signature genes, CCL2, and CXCL10 were determined by the ΔΔCT method. Results were correlated with therapy using prednisone, mycophenolate mofetil, and hydroxychloroquine and analyzed by Wilcoxon/Kruskal-Wallis test and Fisher's exact test.

Results: CCL2 and CXCL10 were significantly higher in untreated patients compared to treated patients, however, in high STAT1 patient visits there is no significant difference between treated and untreated patients' visits. When comparing linear regression fits of interferon (IFN) score with CCL2 and CXCL10, untreated patients and high STAT1 patients displayed significantly higher slopes compared to treated patients. There was no significant difference between the slopes of high STAT1 and untreated patients indicating that CCL2 and CXCL10 were correlated with type-I IFN in high STAT1 patients similar to that in untreated patients. CCL2 and CXCL10 levels in the high STAT1 subset remained high in treated patient visits compared to those of the low STAT1 subset.

Conclusions: Among the biomarkers analyzed, only CCL2 and CXCL10 showed significantly reduced levels in treated compared to untreated SLE patients. STAT1, CCL2, and CXCL10 are potentially useful indicators of therapeutic action in SLE patients. Further work is needed to determine whether high STAT1 levels convey resistance to therapies commonly used to treat SLE and whether STAT1 inhibitors may have therapeutic implication for these patients.

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Figures

Figure 1

Figure 1

Comparison in the levels of various clinical parameters and biomarkers in SLE patient visits with or without treatment. (A) Disease activity, (B-C) complement levels, (D) anti-dsDNA antibody levels, (E) IFN score, (F) ADAR, (G) STAT1, (H) CCL2, (I) CXCL10, (J) miR-146a, (K) pri-miR-146a, and (L). TNFα in treated (Tx) and untreated (UTX) SLE patient visits as well as healthy donors (HD). Data are presented as box plot. All groups were compared among each other and only significant P values are shown indicating each specific comparison. Average trend lines for high STAT1 (blue) and low STAT1 (red) patient visit subsets are also shown for comparison. Detail comparison between high and low STAT1 subsets are shown in Additional file 1: Figure S2. ADAR, adenosine deaminase acting on RNA; CCL2, C-C motif chemokine ligand 2; CXCL10, C-X-C motif chemokine 10; dsDNA, double-stranded DNA; IFN, interferon; SLE, systemic lupus erythematosus; STAT1, signal transducer and activator of transcription 1; TNFα, tumor necrosis factor alpha.

Figure 2

Figure 2

Comparison of the levels of various biomarkers in the SLE patient visits with prednisone (PDN) therapy versus untreated. Data were analyzed as in Figure 1 except only patients receiving PDN in the treated patient population were included. (A) Disease activity, (B-C) complement levels, (D) anti-dsDNA antibody levels, (E) IFN score, (F) ADAR, (G) STAT1, (H) CCL2, (I) CXCL10, (J) miR-146a, (K) pri-miR-146a, and (L) TNFα in treated (Tx) and untreated (UTX) SLE patient visits as well as healthy donors (HD). Data are presented as box plot. All groups were compared among each other and only significant P values are shown indicating each specific comparison. Average trend lines for high STAT1 (blue) and low STAT1 (red) patient visit subsets are also shown for comparison. Detail comparison between high and low STAT1 subsets are shown in Additional file 1: Figure S3. ADAR, adenosine deaminase acting on RNA; CCL2, C-C motif chemokine ligand 2; CXCL10, C-X-C motif chemokine 10; dsDNA, double-stranded DNA; IFN, interferon; SLE, systemic lupus erythematosus; STAT1, signal transducer and activator of transcription 1; TNFα, tumor necrosis factor alpha.

Figure 3

Figure 3

Comparison of the levels of various biomarkers in the SLE patients visits with hydroxychloroquine (HCQ) therapy versus untreated. Data were analyzed as in Figure 1 except only patients receiving HCQ in the treated patient population were included. (A) Disease activity, (B-C) complement levels, (D) anti-dsDNA antibody levels, (E) IFN score, (F) ADAR, (G) STAT1, (H) CCL2, (I) CXCL10, (J) miR-146a, (K) pri-miR-146a, and (L) TNFα in treated (Tx) and untreated (UTX) SLE patient visits as well as healthy donors (HD). Data are presented as box plot. All groups were compared among each other and only significant P values are shown indicating each specific comparison. Average trend lines for high STAT1 (blue) and low STAT1 (red) patient visit subsets are also shown for comparison. Detail comparison between high and low STAT1 subsets are shown in Additional file 1: Figure S4. ADAR, adenosine deaminase acting on RNA; CCL2, C-C motif chemokine ligand 2; CXCL10, C-X-C motif chemokine 10; dsDNA, double-stranded DNA; IFN, interferon; SLE, systemic lupus erythematosus; STAT1, signal transducer and activator of transcription 1; TNFα, tumor necrosis factor alpha.

Figure 4

Figure 4

Comparison of the levels of various biomarkers in the SLE patients visits with mycophenolate mofetil (MMF) therapy versus untreated. Data were analyzed as in Figure 1 except only patients receiving MMF in the treated patient population were included. (A) Disease activity, (B-C) complement levels, (D) anti-dsDNA antibody levels, (E) IFN Score, (F) ADAR, (G) STAT1, (H) CCL2, (I) CXCL10, (J) miR-146a, (K) pri-miR-146a, and (L) TNFα in treated (Tx) and untreated (UTX) SLE patient visits as well as healthy donors (HD). Data are presented as box plot. All groups were compared among each other and only significant P values are shown indicating each specific comparison. Average trend lines for high STAT1 (blue) and low STAT1 (red) patient visit subsets are also shown for comparison. Detail comparison between high and low STAT1 subsets are shown in Additional file 1: Figure S5. ADAR, adenosine deaminase acting on RNA; CCL2, C-C motif chemokine ligand 2; CXCL10, C-X-C motif chemokine 10; dsDNA, double-stranded DNA; IFN, interferon; SLE, systemic lupus erythematosus; STAT1, signal transducer and activator of transcription 1; TNFα, tumor necrosis factor alpha.

Figure 5

Figure 5

Comparison of the levels of various biomarkers in the SLE patient visits given PDN, MMF, and HCQ therapy at high or low dosage. Differences between doses of PDN (D, G, J, M), MMF (B, E, H, K, N), and HCQ (C, F, I, L, O) were not significant with the exception of SLEDAI (A), and in fact SLEDAI scores were higher for PDN dose of 20 to 60 mg/day compared to the 2 to 18 mg/day of dose. HCQ, hydroxychloroquine; MMF, mycophenolate mofetil; PDN, prednisone; SLE, systemic lupus erythematosus; SLEDAI, SLE disease activity index.

Figure 6

Figure 6

Association between CCL2, IFN score, and therapy. (A) The relationship of CCL2 versus IFN score presented as a slope was analyzed in untreated (UTX, black) and treated SLE patient visits (Tx, green). Similar analyses were carried out for PDN-treated (C), MMF-treated (E), and HCQ-treated patient visits (G) as well as for high STAT1 (blue) and low STAT1 (red) for Tx (B), PDN-treated (D), MMF-treated (F), and HCQ-treated patient visits (H). CCL2, C-C motif chemokine ligand 2; HCQ, hydroxychloroquine; IFN, interferon; MMF, mycophenolate mofetil; PDN, prednisone; SLE, systemic lupus erythematosus; SLEDAI, SLE disease activity index; STAT1, signal transducer and activator of transcription 1.

Figure 7

Figure 7

Association between CXCL10, IFN score, and therapy. Data were analyzed as in Figure 6 except that CCL2 was substituted by CXCL10. (A) The relationship of CCL2 versus IFN score presented as a slope was analyzed in untreated (UTX, black) and treated SLE patient visits (Tx, green). Similar analyses were carried out for PDN-treated (C), MMF-treated (E), and HCQ-treated patient visits (G) as well as for high STAT1 (blue) and low STAT1 (red) for Tx (B), PDN-treated (D), MMF-treated (F), and HCQ-treated patient visits (H). CCL2, C-C motif chemokine ligand 2; CXCL10, C-X-C motif chemokine 10; HCQ, hydroxychloroquine; IFN, interferon; MMF, mycophenolate mofetil; PDN, prednisone; SLE, systemic lupus erythematosus; SLEDAI, SLE disease activity index; STAT1, signal transducer and activator of transcription 1.

Figure 8

Figure 8

The expression of CCL2 and CXCL10 in high versus low STAT1 patient subsets with individual and combined therapy. (A) CCL2 levels in HD (healthy donor), untreated (UTX), and other patient visits under treatment with different combinations of PDN, MMF, and HCQ were plotted (black bars). Only significant differences comparing each treatment group to either HD or UTX are indicated as black lines with P value shown. Data segregating into high STAT1 (blue line) and low STAT1 (red line) subsets are also shown and significant differences for these subsets are indicated by green asterisks. (B) CXCL10 data were analyzed similarly. CCL2, C-C motif chemokine ligand 2; CXCL10, C-X-C motif chemokine 10; HCQ, hydroxychloroquine; HD, healthy donor; IFN, interferon; MMF, mycophenolate mofetil; PDN, prednisone; SLE, systemic lupus erythematosus; SLEDAI, SLE disease activity index; STAT1, signal transducer and activator of transcription 1.

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