Introduction
Fibrostenotic Crohn’s disease (FSCD) is a common, treatment-refractory complication, driven by inflammation dependent and independent fibrotic remodeling. The absence of accurate biomarkers limits precision medicine and treatment stratification.
Aims & Methods
We aimed to develop a clinically actionable proteomic signature by integrating full-thickness intestinal resection tissue with parallel matched longitudinal serum profiling to enable non-invasive patient stratification and monitoring. In this prospective study, 62 patients with FSCD, defined according to CONSTRICT criteria, undergoing intestinal resection were included. 48 CD patients with luminal disease without stenosis served as a control. Disease activity and extent was assessed using MR enterography and ileocolonoscopy. High-throughput proteomic profiling was performed using the Olink Explore HT5400 platform (Olink Proteomics, Uppsala, Sweden) on snap frozen full-thickness intestinal tissue samples (ileal (proximal) & colonic (distal) resection margins plus the most stenotic area) and serum from the same patient. Serum samples were collected at baseline (time of resection) and at 1, 3, 6, and 12 months of follow-up. Controls did not undergo resection. Linear models, differential expression analyses, and temporal trajectory analyses were applied to identify discriminatory protein signatures, which were subsequently validated in internal and external datasets.
Results
Proteomic analyses on full-thickness FSCD intestinal resection tissues were performed comparing most stenotic areas to proximal & distal margins and adjusted for sex and smoking status (FDR < 0.01, fold change > 1.5). Out of 5,400 proteins, we identified 1,349 differentially expressed proteins in stenotic tissue (896 upregulated and 453 downregulated). Parallel serum proteomic analysis comparing FSCD patients with luminal disease only (FDR < 0.05, fold change > 1.3), identified 276 differentially expressed proteins (261 upregulated and 15 downregulated). A total of 91 proteins overlapped between tissue and serum analyses to be associated with FSCD. Among these, extracellular matrix–related proteins such as THBS2, SPON2, and RSPO1 were consistently elevated in FSCD across both tissue and serum at baseline. The same proteins exhibited significant reduction in serum following surgical resection, but still remained elevated compared to luminal CD serum samples throughout 12 month follow-up. No significant differences for THBS2, SPON2, and RSPO1 were observed between macroscopically normal proximal and distal resection margins, providing a robust internal control.
Conclusion
These findings demonstrate association of distinct proteins with FSCD between serum and intestinal tissue. Those proteins decrease in serum upon stricture resection and hence are likely relevant. This highlights a coordinated signature of matrix remodeling, immune activation, and dysregulated tissue repair, with potential as FSCD phenotype-specific biomarkers.
Disclosure
AP has received research grants from Janssen-Cilag, Takeda, Tillotts Pharma, AbbVie, MSD, Ferring, and Johnson & Johnson Innovative Medicine; personal fees from Janssen, Takeda, Tillotts Pharma, Ferring, Bristol Myers Squibb, and AbbVie; and consultancy fees and advisory board honoraria from Agomab, Eli Lilly, Bristol Myers Squibb, Johnson & Johnson, and Takeda. SL, KP, SV, and DR are employees of Johnson & Johnson Innovative Medicine, Immunology, Translational Science. MP, CL, and JR declare no conflicts of interest. LBR has received consultancy fees for advisory board participation from Sakura Finetek Europe. IG reports research grants from Pharmacosmos and Intuitive Surgical, as well as personal fees for congress attendance and presentations from Pharmacosmos, Intuitive Surgical, and Johnson & Johnson; none of these are related to the present work. JB has received research grants from AbbVie, Janssen-Cilag, MSD, Takeda, Tillotts Pharma, Bristol Myers Squibb, and Novo Nordisk Foundation; and personal fees from AbbVie, Janssen-Cilag, Celgene, MSD, Pfizer, Takeda, Tillotts Pharma, Bristol Myers Squibb, Samsung Bioepis, Pharmacosmos, Ferring, Galapagos, Eli Lilly, Dr. Falk Pharma, Celltrion, Zealand Pharma, and Orion Pharma. JBS has received research grants from Takeda, Janssen, the Danish Research Council, and the Capital Region Denmark, and serves as national coordinator for studies sponsored by AbbVie, Arena Pharmaceuticals, Eli Lilly, and Boehringer Ingelheim; none of these are related to the present work. FR reports consultancy and advisory board participation for multiple companies, including Adnovate, Agomab, Allergan, AbbVie, Boehringer Ingelheim, Celgene/BMS, CDISC, Cowen, Galmed, Genentech, Gilead, Gossamer, Guidepoint, Helmsley, Index Pharma, Janssen, Koutif, Mestag, Metacrine, Morphic, Organovo, Origo, Pfizer, Pliant, Prometheus Biosciences, Receptos, RedX, Roche, Samsung, Surmodics, Surrozen, Takeda, Techlab, Theravance, Thetis, UCB, Ysios, and 89Bio; and has received research funding from the National Institutes of Health, Helmsley Charitable Trust, Crohn’s and Colitis Foundation, UCB, Pliant, Bristol Myers Squibb, AbbVie, Pfizer, Boehringer Ingelheim, Morphic, and the Kenneth Rainin Foundation.