Background Interstitial lung disease (ILD) is a life-threatening complication in systemic sclerosis (SSc). Substantial research progress has identified distinct genomic and molecular subtypes in SSc-ILD and brought molecular targeted therapies within reach. However, personalized medicine approaches are still lacking since clinically applicable tools for individualized patient stratification are not yet available.
Objectives To assess the possibility of imaging molecular targets as a biomarker for stage-dependent assessment of ILD in the mouse model of bleomycin-induced lung fibrosis.
Methods Expression of integrin αvβ3 and folate receptor β (FR-β) was analyzed in lung tissues from patients with idiopathic pulmonary fibrosis (IPF), SSc-ILD, and healthy controls as well as from bleomycin treated mice and saline treated controls using immunohistochemistry and qPCR (n=5–11). SPECT (Single Photon Emission Computed Tomography) or PET (Positron Emission Tomography) was performed at days 3, 7, and 14 after bleomycin instillation using the integrin αvβ3-specific 177Lu-c(RGDfK)-ligand and the FR-β-specific 18F-Azafol. Additionally, 18F-FDG-PET and high resolution CT (HRCT) scans were performed. The specific lung uptake of the radiotracers over time was assessed by ex vivo SPECT or PET/CT scans and biodistribution studies.
Results Expression of FR-β was significantly increased at the mRNA and protein level of SSc-ILD patients as compared to healthy subjects (p<0.05), whereas only its gene expression was upregulated in IPF patients (p<0.01). Integrin αvβ3 was increased at the protein level of both SSc-ILD and IPF patients (p<0.05), while its mRNA expression was not significantly altered.
Similarly, in lungs of bleomycin treated mice, but not of controls, FR-β expression was increased time-dependently at the mRNA and protein level with higher expression at day 3 and day 7, the inflammatory stages of bleomycin-induced lung fibrosis (p<0.05). In contrast, expression of integrin αvβ3 was upregulated at day 7 and day 14 at the protein, but not at the mRNA level in bleomycin treated mice, and thus not only in the inflammatory but also in the fibrotic stages (p<0.05). 18F-FDG-PET and HRCT detected time-dependent changes of metabolic activity and ILD morphology in bleomycin treated mice. However, compared with these routinely employed unselective imaging techniques, molecular targeted imaging of integrin αvβ3 and FR-β successfully visualized ILD and discriminated lung inflammation and/or fibrosis in a time-dependent manner and in correspondence with the expression changes at the tissue level. The specific lung uptake of 177Lu-c(RGDfK)-ligand and 18F-Azafol as compared with the unspecific uptake of 18F-FDG in diseased lungs over time was demonstrated by biodistribution studies and ex vivo SPECT or PET/CT scans.
Conclusions Our data suggest that stage-dependent visualization of ILD with radiotracers that target key markers of lung inflammation and/or fibrosis shows promise for clinical application. As opposed to unselective imaging techniques such as 18F-FDG-PET and HRCT, the introduction of specific imaging biomarkers for individualized management of SSc-ILD patients could represent the first step towards precision medicine.
Disclosure of Interest J. Schniering Grant/research support from: Swiss National Science Foundation (S-85605–02–01), M. Benešová Grant/research support from: Swiss National Science Foundation (S-85605–02–01), M. Brunner: None declared, C. Feghali-Bostwick: None declared, R. Schibli Grant/research support from: Merck & Cie, O. Distler Grant/research support from: Actelion, Bayer, Boehringer Ingelheim, Pfizer, Sanofi; Patent licensed: mir-29 for the treatment of systemic sclerosis, Consultant for: 4 D Science, Actelion, Active Biotec, Bayer, BiogenIdec, BMS, Boehringer Ingelheim, ChemomAb, EpiPharm, espeRare foundation, Genentech/Roche, GSK, Inventiva, Lilly, medac, Mepha, MedImmune, Mitsubishi Tanabe Pharma, Pharmacyclics, Pfizer, Sanofi, Serodapharm, Sinoxa, Speakers bureau: AbbVie, iQone Healthcare, Mepha, C. Müller Grant/research support from: Merck & Cie, B. Maurer Grant/research support from: AbbVie, Protagen, EMDO, Novartis, Pfizer, Roche, Actelion; Patent licensed: mir-29 for the treatment of systemic sclerosis