913cba85c66027c3f6192cdd4a16bcc1 306342.pdf 6941d2a4e7741d78619fb9f53c839a356b02df12 306342.pdf 13cb2a3f5d39e2dee75d6757b08a4d966eb9c9f1abe47fc14b62312fde79ea55 306342.pdf Title: Kinetics of IFN-Induced Cytokines and Development of Immune-Related Adverse Events in Patients Receiving PD-(L)1 Inhibitors Subject: Immune checkpoint inhibitors (ICI) have the potential to trigger unpredictable immune-related adverse events (irAEs), which can be severe. The underlying mechanisms of these events are not fully understood. As PD-L1 is upregulated by IFN, the heightened immune activation resulting from PD-1/PD-L1 inhibition may enhance the IFN response, triggering the expression of IFN-inducible genes and contributing to irAE development and its severity. In this study, we investigated the interplay between irAEs and the expression of IFN-inducible chemokines and cytokines in 134 consecutive patients with solid tumours treated with PD-(L)1 inhibitors as monotherapy or in combination with chemotherapy or other immunotherapy agents. We compared the plasma levels of IFN-associated cytokines (CXCL9/10/11, IL-18, IL-10, IL-6 and TGF) at various time points (at baseline, at the onset of irAE and previous to irAE onset) in three patient groups categorized by irAE development and severity: patients with serious irAEs, mild irAEs and without irAEs after PD-(L)1 inhibitors. No differences were observed between groups at baseline. However, patients with serious irAEs exhibited significant increases in CXCL9/10/11, IL-18 and IL-10 levels at the onset of the irAE compared to baseline. A network analysis and correlation patterns highlighted a robust relationship among these chemokines and cytokines at serious-irAE onset. Combining all of the analysed proteins in a cluster analysis, we identified a subgroup of patients with a higher incidence of serious irAEs affecting different organs or systems. Finally, an ROC analysis and a decision tree model proposed IL-18 levels 807 pg/mL and TGF levels 114 pg/mL as predictors for serious irAEs in 90% of cases. In conclusion, our study elucidates the dynamic changes in cytokine profiles associated with serious irAE development during treatment with PD-(L)1 inhibitors. The study’s findings offer valuable insights into the intricate IFN-induced immune responses associated with irAEs and propose potential predictive markers for their severity. Keywords: interferon; cytokines; immune checkpoint inhibitors; anti-PD-(L)1 inhibitors; immune-related adverse events (irAEs); predictive biomarkers Author: Leticia Alserawan, Maria Mulet, Geòrgia Anguera, Mariona Riudavets, Carlos Zamora, Rubén Osuna-Gómez, Jorgina Serra-López, Andrés Barba Joaquín, Ivana Sullivan, Margarita Majem and Silvia Vidal Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Wed May 1 09:32:30 2024 CEST ModDate: Fri May 3 05:46:17 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 16 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2493430 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- AKXRSZ+EURB10 Type 1 Builtin yes yes yes 10 0 RNXXBO+VnURWPalladioL-Bold Type 1 Custom yes yes yes 15 0 GOCQIR+EURM10 Type 1 Builtin yes yes 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Identity-H yes yes yes 256 0 BCDJEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 263 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 279 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 283 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 287 0 BCDHEE+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 291 0 BCDIEE+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 298 0 BCDJEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 305 0 BCDLEE+SegoeUI CID TrueType Identity-H yes yes yes 312 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-01-28 02:39:56 CET RepresentationInformation: 306342.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-01-27 14:04:37 CET Size: 2493430 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 509 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Patient Inclusion and Clinical Assessment Destination: subsection.2.1 Item: Title: Sample Collection Destination: subsection.2.2 Item: Title: Determination of Plasmatic Concentration of Cytokines Destination: subsection.2.3 Item: Title: Statistical Analysis Destination: subsection.2.4 Item: Title: Results Destination: section.3 Children: Item: Title: Patient Characteristics and Types of irAEs Destination: subsection.3.1 Item: Title: Changes in the Levels of Cytokines during ICI Treatment According to irAE Severity Destination: subsection.3.2 Item: Title: Interplay of Cytokines in irAE Samples, According to the Severity of the irAE Destination: subsection.3.3 Item: Title: Clustering of irAE Samples Based on Cytokines and Their Association with Severity and Type of irAE Destination: subsection.3.4 Item: Title: Pre-irAE Cytokine Levels as Predictive Markers for the Development of Serious irAEs Destination: subsection.3.5 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: appendix.A. Info: Title: Kinetics of IFN-Induced Cytokines and Development of Immune-Related Adverse Events in Patients Receiving PD-(L)1 Inhibitors Author: Leticia Alserawan, Maria Mulet, Geòrgia Anguera, Mariona Riudavets, Carlos Zamora, Rubén Osuna-Gómez, Jorgina Serra-López, Andrés Barba Joaquín, Ivana Sullivan, Margarita Majem and Silvia Vidal Subject: Immune checkpoint inhibitors (ICI) have the potential to trigger unpredictable immune-related adverse events (irAEs), which can be severe. The underlying mechanisms of these events are not fully understood. As PD-L1 is upregulated by IFN, the heightened immune activation resulting from PD-1/PD-L1 inhibition may enhance the IFN response, triggering the expression of IFN-inducible genes and contributing to irAE development and its severity. In this study, we investigated the interplay between irAEs and the expression of IFN-inducible chemokines and cytokines in 134 consecutive patients with solid tumours treated with PD-(L)1 inhibitors as monotherapy or in combination with chemotherapy or other immunotherapy agents. We compared the plasma levels of IFN-associated cytokines (CXCL9/10/11, IL-18, IL-10, IL-6 and TGF) at various time points (at baseline, at the onset of irAE and previous to irAE onset) in three patient groups categorized by irAE development and severity: patients with serious irAEs, mild irAEs and without irAEs after PD-(L)1 inhibitors. No differences were observed between groups at baseline. However, patients with serious irAEs exhibited significant increases in CXCL9/10/11, IL-18 and IL-10 levels at the onset of the irAE compared to baseline. A network analysis and correlation patterns highlighted a robust relationship among these chemokines and cytokines at serious-irAE onset. Combining all of the analysed proteins in a cluster analysis, we identified a subgroup of patients with a higher incidence of serious irAEs affecting different organs or systems. Finally, an ROC analysis and a decision tree model proposed IL-18 levels 807 pg/mL and TGF levels 114 pg/mL as predictors for serious irAEs in 90% of cases. In conclusion, our study elucidates the dynamic changes in cytokine profiles associated with serious irAE development during treatment with PD-(L)1 inhibitors. The study’s findings offer valuable insights into the intricate IFN-induced immune responses associated with irAEs and propose potential predictive markers for their severity. 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