Multiplex serum protein analysis reveals potential mechanisms and markers of response to hyperimmune caprine serum in systemic sclerosis

Abstract

BackgroundHyperimmune caprine serum (HICS) is a novel biological therapy with potential benefit for skin in established diffuse cutaneous systemic sclerosis. Here we report multiplex protein analysis of blood samples from a placebo-controlled phase II clinical trial and explore mechanisms of action and markers of response.MethodsPatients were treated with HICS (n = 10) or placebo (n = 10) over 26 weeks, with follow-up open-label treatment to 52 weeks in 14 patients. Serum or plasma samples at baseline, 26 and 52 weeks were analysed using multiplex or individual immunoassays for 41 proteins. Patterns of change were analysed by clustering using Netwalker 1.0, Pearson coefficient and significance analysis of microarrays (SAM) correction.ResultsCluster analysis, SAM multiplex testing and paired comparison of individual analytes identified proteins that were upregulated or downregulated during treatment with HICS. There was upregulation of the hypothalamo-pituitary-adrenal axis after HICS treatment evidenced by increases in α-MSH and ACTH in cases treated with HICS. Interestingly, significant increase in PIIINP was associated with HICS treatment and improved MRSS suggesting that this may be a marker of extracellular matrix turnover. Other relevant factors reduced in HICS-treated patients compared with controls, although not reaching statistical significance included COMP, CCL2, IL6, TIMP2, Fractalkine and TGFβ1 levels.ConclusionsOur results suggest mechanisms of action for HICS, including upregulation of α-MSH, that has been shown to be anti-fibrotic in preclinical models, and possible markers to be included in future trials targeting skin in diffuse cutaneous systemic sclerosis.Trial registrationEudract, No. 2007-003122-24. ClinTrials.gov, No. NCT00769028. Registered 7 October 2008.Electronic supplementary materialThe online version of this article (doi:10.1186/s13075-017-1252-x) contains supplementary material, which is available to authorized users.

European multicentre study validates enhanced liver fibrosis test as biomarker of fibrosis in systemic sclerosis

“…Indeed, ELF score and the single markers were significantly higher in patients with dcSSc, severe skin involvement, fibrosis on chest HRCT scan, abnormal pulmonary function and DLCO. The higher PIIINP level of patients on immunosuppressive therapy is consistent with published data and may reflect a higher level of ECM turnover (11). By contrast, ELF and the single markers showed no difference in patients with digital ulcers or with specific videocapillaroscopy patterns compared to patients without signs of severe vasculopathy.…”

Challenges in systemic sclerosis trial design

“…In addition, the 2 gene skin score biomarker was significantly reduced agree treatment with anti TGFbeta monoclonal antibody. In a trial of the novel biological agent HICS there were changes in serum markers that reflected the likely mechanism of action by stimulating the hypothalamopituitary axis and upregulating MSH which has antifibrotic activity in preclinical models [6]. There was a link between markers of fibrosis and clinical improvement in MRSS over 26 weeks in this clinical trial [7].…”

Interleukin-6 and Related Proteins as Biomarkers in Systemic Sclerosis

“…This was not seen in a study of imatinib, which instead showed no evidence of benefit for this treatment and evidence of substantial toxicity (31). In a trial of hyperimmune caprine serum, skin scores did improve and IL-6 was among the serum cytokines whose levels decreased during therapy, thereby indicating it as a possible marker of skin change (32).…”