A biomarker signature to predict complete response to itacitinib and corticosteroids in acute graft-versus-host disease

Michael Pratta, Sophie Paczesny, Gerard Socie, Natalie Barkey, Hao Liu, Sherry Owens, Michael C Arbushites, Mark A Schroeder, Michael D Howell, Michael Pratta, Sophie Paczesny, Gerard Socie, Natalie Barkey, Hao Liu, Sherry Owens, Michael C Arbushites, Mark A Schroeder, Michael D Howell

Abstract

A broad proteomic analysis was conducted to identify and evaluate candidate biomarkers potentially predictive of response to treatment with an oral selective Janus kinase 1 (JAK1) inhibitor, itacitinib, in acute graft-versus-host disease (GVHD). Plasma samples from 25 participants (identification cohort; NCT02614612) were used to identify novel biomarkers that were tested in a validation cohort from a placebo-controlled, randomised trial (n = 210; NCT03139604). The identification cohort received corticosteroids plus 200 or 300 mg itacitinib once daily. The validation cohort received corticosteroids plus 200 mg itacitinib once daily or placebo. A broad proteomic analysis was conducted using a proximity extension assay. Baseline and longitudinal comparisons were performed with unpaired t-test and one-way analysis of variance used to evaluate biomarker level changes. Seven candidate biomarkers were identified. Monocyte-chemotactic protein (MCP)3, pro-calcitonin/calcitonin (ProCALCA/CALCA), together with a previously identified prognostic acute GVHD biomarker, regenerating islet-derived protein (REG)3A, stratified complete responders from non-responders (participants with progressive disease) to itacitinib, but not placebo, potentially representing predictive biomarkers of itacitinib in acute GVHD. ProCALCA/CALCA, suppressor of tumorigenicity (ST)2, and tumour necrosis factor receptor (TNFR)1 were significantly reduced over time by itacitinib in responders, potentially representing response-to-treatment biomarkers. Novel biomarkers have the potential to identify patients with acute GVHD that may respond to itacitinib plus corticosteroid treatment (NCT02614612; NCT03139604).

Keywords: JAK inhibitors; acute graft-versus-host disease; biomarkers; itacitinib.

Conflict of interest statement

Sophie Paczesny has a patent, ‘Methods of detection of graft‐versus‐host disease’ (US 20130115232A1, WO 2013066369A3), licensed to Viracor‐IBT Laboratories, and received honoraria from Incyte, Genentech, and Omeros for advisory board meetings. Gerard Socie received lecture fees from Incyte Corporation and Novartis and honoraria from Novartis for advisory board meeting. Mark A. Schroeder received research funding paid to his institution by companies for which he is a Principal Investigator for a company‐sponsored research study including Genentech Inc., Incyte, Cellect Inc., Fortis, Seattle Genetics, Amgen, Celgene, PDB Incorporated, Genzyme Sanofi, and Janssen; served on advisory boards, and received honoraria and consultant fees from Amgen, Astellas, Dova Pharmaceuticals, FlatIron Inc., Incyte, Partners Therapeutics, Pfizer, and Sanofi Genzyme; and served on the speakers bureau and received honoraria as a consultant for AbbVie, Merck, and Takeda. Michael Pratta, Natalie Barkey, and Michael C. Arbushites are employees of Incyte Research Institute, Incyte Corporation. Hao Liu, Sherry Owens, and Michael D. Howell were employees of Incyte Corporation at the time these studies were undertaken.

© 2022 INCYTE Corporation. British Journal of Haematology published by British Society for Haematology and John Wiley & Sons Ltd.

Figures

FIGURE 1
FIGURE 1
Volcano plot representing differentially expressed proteins at baseline in plasma of CR compared with PD groups in identification cohort. Broad proteomic analysis of plasma samples was conducted by OLINK proteomics (Watertown, MA, USA) using a proximity extension assay as described by the manufacturer. The protein library consists of >1000 proteins. Data are presented as NPX in log2 scale. FC was calculated based on NPX values in CR (n = 10) and PD (n = 6). A negative FC represents proteins downregulated in CR relative to PD; a positive FC represents proteins upregulated in CR relative to PD. In the volcano plot shown, the x‐axis is the log2 transformed FC, calculated as log2(FC), and y‐axis is the negative log p value, calculated as –log10(raw p value). Identity of the novel predictive candidate biomarkers is shown in green, JAK/STAT‐related biomarkers are shown in red, and previously validated prognostic biomarkers are shown in blue. Identity of all other proteins illustrated is listed in Table S2. CALCA, calcitonin; CCL19, C‐C motif chemokine 19; CR, complete responder; CXCL10, C‐X‐C motif chemokine 10; FC, fold change; IL2RA, interleukin 2 receptor alpha chain; IL6, interleukin‐6; IL8, interleukin‐8; JAK, Janus kinase; MCP3, monocyte‐chemotactic protein 3; NPX, normalised protein expression; PD, progressive disease or death; PON3, paraoxonase 3; REG3A, regenerating islet‐derived protein; SCF, kit ligand; ST2, suppressor of tumorigenicity 2; STAT, signal transducer and activator of transcription; TNFR1, tumour necrosis factor receptor 1; TNFRSF6B, TNF receptor superfamily member 6b.
FIGURE 2
FIGURE 2
Baseline levels of novel predictive candidate biomarkers, JAK/STAT‐related biomarkers, and previously validated prognostic biomarkers in identification cohort. Baseline plasma samples from CR and PD participants (or participants who died) and from participants with an intermediate response, including VGPR and PR, were assayed for biomarker levels by proximal extension assay. Statistical analysis was by one‐way ANOVA. Significance was conferred when p < 0.05. The lines over particular cohorts indicate comparison of those cohorts with p values indicated. ANOVA, analysis of variance; CCL19, C‐C motif chemokine 19; CR, complete responder; CXCL10, C‐X‐C motif chemokine 10; IL2RA, interleukin 2 receptor alpha chain; IL6, interleukin 6; IL8, interleukin 8; JAK, Janus kinase; MCP3, monocyte‐chemotactic protein 3; nat, natural; PD, progressive disease; PON3, paraoxonase 3; ProCALCA/CALCA, pro‐calcitonin/calcitonin; PR, partial responder; REG3A, regenerating islet‐derived protein; SCF, kit ligand; ST2, suppressor of tumorigenicity 2; STAT, signal transducer and activator of transcription; TNFR1, tumour necrosis factor receptor 1; TNFRSF6B, TNF receptor superfamily member 6b; VGPR, very good partial responder; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; CR, blue; PD/death, black; VGPR/PR, red.
FIGURE 3
FIGURE 3
Comparison of day 28 treatment response between itacitinib versus placebo in validation cohort. Day 28 treatment responses from itacitinib (n = 101) and placebo (n = 109) treatment arms from the validation cohort are presented. PD includes participants whose disease progressed during treatment and/or who died before day 28. The ‘other’ category includes mixed responders, non‐responders, and unclassified participants. CR, complete responder; PD, progressive disease/death; VGPR/PR, very good partial responder/partial responder.
FIGURE 4
FIGURE 4
Baseline levels of candidate predictive biomarkers (A) ProCALCA/CALCA; (B) REG3A; (C) MCP3; (D) ST2 in the validation cohort. Baseline serum samples from itacitinib versus placebo arms of validation cohort were assayed for biomarker levels by proximal extension assay. Statistical analysis was by unpaired t‐test. Significance was conferred when p < 0.05. Itacitinib CR: Blue; Itacitinib PD: Black; placebo CR: Orange; placebo PD: Green. Open symbols represent participants with standard risk; closed symbols represent participants with high‐risk acute GVHD. CR, complete response; GVHD, graft‐versus‐host disease; MCP3, monocyte‐chemotactic protein 3; nat, natural; NS, not significant; PD, progressive disease/death; ProCALCA/CALCA, pro‐calcitonin/calcitonin; REG3A, regenerating islet‐derived protein; ST2, suppressor of tumorigenicity 2; *p < 0.05; **p < 0.01.
FIGURE 5
FIGURE 5
Longitudinal analysis of candidate biomarkers in responders from validation cohort. Serum levels of candidate biomarkers identified in the identification cohort were measured at baseline and at days 7, 14, and 28 (where available) from responders (CR, VGPR, PR) in the validation cohort, and mean ± SEM values are plotted from the itacitinib/corticosteroid combination (n = 74; red circles) versus corticosteroid alone (placebo n = 79; blue squares). Data are shown for each biomarker showing a significant difference between treatment arms. Data were compared at each time point by unpaired t‐test, and significant differences between treatment arms are shown (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). BL, baseline; CR, complete responder; MCP3, monocyte‐chemotactic protein 3; nat, natural; PR, partial responder; ProCALCA/CALCA, pro‐calcitonin/calcitonin; REG3A, regenerating islet‐derived protein; SEM, standard error of the mean; ST2, suppressor of tumorigenicity 2; TNFR1, tumour necrosis factor receptor 1; VGPR, very good partial responder.
FIGURE 6
FIGURE 6
Density plots comparing baseline levels of each candidate biomarker in the identification and validation cohorts. Baseline levels of each candidate biomarker are shown in density plots, where log2 of biomarker concentration was compared with the density of participants at a corresponding biomarker concentration. Baseline levels from identification cohort are shown in blue; baseline levels from the validation cohort are shown in yellow. The markers were classified according to novel predictive candidate biomarkers, JAK/STAT‐related biomarkers, and previously validated prognostic biomarkers. Statistical analysis was by unpaired t‐test. Significance was conferred when p < 0.05. The lines over particular cohorts indicate comparison of those cohorts with p values indicated. CCL19, C‐C motif chemokine 19; CXCL10, C‐X‐C motif chemokine 10; IL2RA, interleukin‐2 receptor alpha chain; IL6, interleukin 6; IL8, interleukin 8; JAK, Janus kinase; MCP3, monocyte‐chemotactic protein 3; NS, not significant; PD, progressive disease; PON3, paraoxonase 3; ProCALCA/CALCA, pro‐calcitonin/calcitonin; REG3A, regenerating islet‐derived protein; SCF, kit ligand; ST2, suppressor of tumorigenicity 2; STAT, signal transducer and activator of transcription; TNFR1, tumour necrosis factor receptor 1; TNFRSF6B, TNF receptor superfamily member 6b. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
FIGURE 7
FIGURE 7
Correlation between protein simple Ella and proximal extension assay in the detection of candidate biomarkers in samples from identification cohort. Plasma samples from CR and PD participants from the identification cohort were tested for (A) ProCALCA/CALCA; (B) REG3A; (C) MCP3; (D) ST2; and (E) TNFR1 by protein simple Ella (x‐axis) and by PEA (y‐axis). Correlation between the two assay platforms is shown for each biomarker. Statistical analysis was performed using Pearson correlation, and r and p values are shown. Significance was conferred when p < 0.05. CR, complete responder; MCP3, monocyte‐chemotactic protein 3; nat, natural; PD, progressive disease/death; PEA, proximal extension assay; ProCALCA/CALCA, pro‐calcitonin/calcitonin; REG3A, regenerating islet‐derived protein; ST2, suppressor of tumorigenicity 2; TNFR1, tumour necrosis factor receptor 1.

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