The online version of this article (doi:10.1186/s13075-017-1252-x) contains supplementary material, which is available to authorized users.
Hyperimmune 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.
Patients 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.
Cluster 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.
Our 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.
Eudract, No. 2007-003122-24. ClinTrials.gov, No. NCT00769028. Registered 7 October 2008.
Additional file 1: Figure S1. Subject progression in the clinical trials with serum sample time points. Schematic indicates how 20 subjects were randomly allocated to active treatment with hyperimmune caprine serum (HICS) or placebo. One patient withdrew from the HICS arm and was not available for follow-up. Other subjects were all followed to 52 weeks and at 26 weeks were offered open-label compassionate HICS. The study blind for 0–26 week treatment was maintained until after 52-week assessment. Serum and plasma samples were available for weeks 0, 26 and 52. Additional screening blood samples (pre-randomisation) were available for quality control purposes. (TIF 132 kb)13075_2017_1252_MOESM1_ESM.tif
Additional file 2: Figure S2. Unsupervised hierarchical cluster analysis for multiplex serum proteins at baseline and 26 weeks for the extended dataset of 26 weeks treatment with HICS. Unsupervised cluster analysis was undertaken to identify any subgroups within the extended dataset of the study cohort at baseline, the end of placebo treatment period, or at 52 weeks based upon the serum levels of multiple protein analytes as described in text. This provided a larger sample size by including 17 subjects treated with HICS over 26 weeks and 13 subjects with 26 weeks of observation on placebo or no active treatment. The same analysis was repeated for serum samples after 26 weeks of treatment with HICS or placebo. The randomly assigned treatment allocation is shown for each subject. Data for baseline samples are shown in panel A together with treatment allocation. After 26 weeks of treatment there were clear changes in the patterns of protein analytes that were spread between the two treatment arms as shown in panel B. (TIF 1373 kb)13075_2017_1252_MOESM2_ESM.tif
Additional file 3: Figure S3. Unsupervised cluster analysis for change in serum proteins from baseline to 26 weeks comparing treatment with hyperimmune caprine serum (HICS) with placebo over 26 weeks and in extended dataset at 52 weeks. (A) Unsupervised cluster analysis of change in protein level during 26-week treatment phase of placebo-controlled trial reveals patterns of change that are reflected in the supervised analysis shown in Fig. 3. Thus, subjects receiving placebo show generally less treatment effect and those treated with HICS show the patterns consistent with the summary changes shown above. (B) Unsupervised cluster analysis is also performed for the extended 52-week dataset that includes subjects moving from placebo to active treatment ( n = 7) in the second 26 weeks and three cases that have no active treatment and were previously on placebo. This complements the presentation of data for baseline and 26 weeks presented in Additional file 1: Figure S1 and shows close congruity for the two 26-week unsupervised heat maps in the extended dataset. (TIF 1444 kb)
Additional file 4: Figure S4. Average change in serum proteins comparing treatment with hyperimmune caprine serum (HICS) or placebo and for MRSS responders versus non-responder at 26 weeks. (A) Average change in serum protein was calculated for each treatment arm over 26 weeks and ranked according to fold change average after HICS treatment. (B) Similar analysis was undertaken for average protein changes in the subjects showing significant improvement of four skin score units and 20% of baseline MRSS score during the trial (responders) or these that did not demonstrate clinical response. These were ranked for the most increased proteins in responder cases. Key proteins that emerged as upregulated (red) or downregulated (blue) for HICS treatment, shown in Fig. 4, are annotated with asterisks. (TIF 761 kb)
Youl BD, Ginsberg L. Goat serum product AIMSPRO® shows promise as an effective treatment in CIDP. London: BSCN meeting, National Hospital; 2004.
Youl BD, Crum J. Clinical improvement in Krabbe’s disease case treated with hyperimmune goat serum product AIMSPRO®. J Neurol Sci. 2005;238:S110.
Youl BD, Angus-Leppan H, Hussein N, Brooman I, Fitzsimons RB. Rapid and sustained response to hyperimmune goat serum product in a patient with Myaesthenia Gravis. J Neurol Sci. 2005;238:S177.
Moore CEG, Hannan R, McIntosh D. In vivo, human peripheral nerve strength duration time constant changes with AIMSPRO® implicate altered sodium channel function as a putative mechanism of action. J Neurol Sci. 2005;238:S238.
Kiernan MC, Burke D, Bostock H. Nerve excitability measures: biophysical basis and use in investigation of peripheral nerve disease use in investigation of peripheral nerve disease. In: Dyck PJ, Thomas PK, editors. Peripheral Neuropathy. 4th ed. Philadelphia: Elsevier Saunders; 2005. p. 113–29. CrossRef
Burke G, Cavey A, Matthews P, Palace J. The evaluation of a novel ‘goat serum’ (AIMSPRO®) in multiple sclerosis. J Neurol Neurosurg Psychiatr. 2005;76:1326.
Youl BD, White SDT, McIntosh D, Cadogan M, Dalgleish AG, Ginsberg L. Hyperimmune serum reverses conduction block in demyelinated human optic nerve and peripheral nerve fibres. J Neurol Neurosurg Psychiatr. 2004;76:615.
Youl BD, Orrell R. Goat serum product AIMSPRO® produces sustained improvement in muscle power in a patient with fascioscapulohumeral dystrophy. J Neurol Sci. 2005;238:S169.
Chung L, Denton CP, Distler O, Furst DE, Khanna D, Merkel PA. Clinical trial design in scleroderma: where are we and where do we go next? Clin Exp Rheumatol. 2012;30(2 Suppl 71):S97–102. PubMed
Khanna D, Furst DE, Allanore Y, Bae S, Bodukam V, Clements PJ, et al. Twenty-two points to consider for clinical trials in systemic sclerosis, based on EULAR standards. Rheumatology (Oxford). 2015;54(1):144–51. CrossRef
Chakravarty EF, Martyanov V, Fiorentino D, Wood TA, Haddon DJ, Jarrell JA, et al. Gene expression changes reflect clinical response in a placebo-controlled randomized trial of abatacept in patients with diffuse cutaneous systemic sclerosis. Arthritis Res Ther. 2015;17:159. CrossRefPubMedPubMedCentral
Takehara K, Ihn H, Sato S. A randomized, double-blind, placebo-controlled trial: intravenous immunoglobulin treatment in patients with diffuse cutaneous systemic sclerosis. Clin Exp Rheumatol. 2013;31(2 Suppl 76):151–6. PubMed
Raja J, Nihtyanova SI, Murray CD, Denton CP, Ong VH. Sustained benefit from intravenous immunoglobulin therapy for gastrointestinal involvement in systemic sclerosis. Rheumatology (Oxford). 2016;55(1):115–9. CrossRef
Clark KE, Etomi O, Denton CP, Ong VH, Murray CD. Intravenous immunogobulin therapy for severe gastrointestinal involvement in systemic sclerosis. Clin Exp Rheumatol. 2015;33(4 Suppl 91):S168–70. PubMed
Zhang Z, Ma J, Yao K, Yin J. Alpha-melanocyte stimulating hormone suppresses the proliferation of human tenon’s capsule fibroblast proliferation induced by transforming growth factor beta 1. Mol Biol (Mosk). 2012;46(4):628–33.
Kokot A, Sindrilaru A, Schiller M, Sunderkotter C, Kerkhoff C, Eckes B, et al. alpha-melanocyte-stimulating hormone suppresses bleomycin-induced collagen synthesis and reduces tissue fibrosis in a mouse model of scleroderma: melanocortin peptides as a novel treatment strategy for scleroderma? Arthritis Rheum. 2009;60(2):592–603. CrossRefPubMed
- Multiplex serum protein analysis reveals potential mechanisms and markers of response to hyperimmune caprine serum in systemic sclerosis
Kristina E. N. Clark
Christopher P. Denton
- BioMed Central
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