22eccad120fba9ad1c77d7b7ce08cbae 293371.pdf f8adf9b6ed23d9012934efda7495567335fcb0d8 293371.pdf 184f8c8ee11e959a763ad28c681afd74f014559457d94cdb52711df35091172a 293371.pdf Title: Potential Role of Circulating PD-L1+ Leukocytes as a Predictor of Response to Anti-PD-(L)1 Therapy in NSCLC Patients Subject: PD-(L)1 inhibitors are part of the treatment strategy for non-small cell lung cancer (NSCLC) although its efficacy is limited to certain patients. Our study aimed to identify patients who might benefit from anti-PD-(L)1 inhibitors by analyzing the PD-L1 expression on circulating leukocytes and its evolution during treatment. One hundred thirteen NSCLC patients, according to their radiological response after 10–12 weeks of treatment, were classified into responders, stable, and progressive disease. Percentages of circulating PD-L1+ leukocytes, PD-L1+ platelets (PLTs), and leukocyte-PLT complexes were assessed using flow cytometry, and plasma concentrations of soluble immunomodulatory factors were quantified by ELISA. Responders exhibited significantly higher pre-treatment percentages of PD-L1+ neutrophils, PD-L1+ CD14+ cells, and PD-L1+ PLTs than progressors. The percentages of these populations decreased in responders post-treatment, contrasting with stables and progressors. PLTs notably contributed to PD-L1 expression in CD14+ cells and neutrophils. Plasma cytokine analysis revealed baseline differences only in IL-17 concentration among groups, whereas network analyses highlighted distinct association patterns between plasma molecules and PD-L1+ leukocytes after 10–12 weeks of treatment. Our findings suggest that pre-treatment assessment of circulating PD-L1+ neutrophils, PD-L1+ CD14+ cells, and PD-L1+ PLTs may be helpful in identifying NSCLC patients who are potential candidates for anti-PD-(L)1 therapy. Keywords: NSCLC; immunotherapy; PD-L1+ neutrophils; PD-L1+ CD14+ cells; PDL-1+ platelets Author: Georgia Anguera, Maria Mulet, Carlos Zamora, Rubén Osuna-Gómez, Andrés Barba, Ivana Sullivan, Jorgina Serra-López, Elisabet Cantó, Silvia Vidal and Margarita Majem Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Fri Apr 26 15:20:17 2024 CEST ModDate: Fri Apr 26 15:25:56 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2881292 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HJOMHV+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 VOBBLV+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 ZHZGAQ+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 FVSCKO+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 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no 941 0 LHALAJ+PalatinoLinotype-Roman TrueType MacRoman yes yes no 944 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-06-06 02:43:46 CEST RepresentationInformation: 293371.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-06-05 12:09:55 CEST Size: 2881292 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 1209 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 Cohort and Sample Collection Destination: subsection.2.1 Item: Title: Evaluation of Response to Immunotherapy by Radiological Assessment Destination: subsection.2.2 Item: Title: Immunophenotyping Destination: subsection.2.3 Item: Title: Flow Cytometry Analysis Destination: subsection.2.4 Item: Title: Determination of Soluble Mediators Destination: subsection.2.5 Item: Title: Statistical Analysis Destination: subsection.2.6 Item: Title: Results Destination: section.3 Children: Item: Title: Patients’ Clinicopathological Characteristics Destination: subsection.3.1 Item: Title: Pretreatment PD-L1 Expression in Leukocytes and Platelets Destination: subsection.3.2 Item: Title: Progressive Changes in PD-L1 Expression on Leukocytes and Platelets Destination: subsection.3.3 Item: Title: Kinetics of PD-L1+ CD14+ Cells and Neutrophils with or without Bound Platelets Destination: subsection.3.4 Item: Title: Determination of Plasma Soluble Immunomodulatory Factors’ Concentrations and Network Analysis Destination: subsection.3.5 Item: Title: Multinomial Logistic Regression Model to Predict Response to Anti-PD-(L)1 Destination: subsection.3.6 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: appendix.A. Info: Title: Potential Role of Circulating PD-L1+ Leukocytes as a Predictor of Response to Anti-PD-(L)1 Therapy in NSCLC Patients Author: Georgia Anguera, Maria Mulet, Carlos Zamora, Rubén Osuna-Gómez, Andrés Barba, Ivana Sullivan, Jorgina Serra-López, Elisabet Cantó, Silvia Vidal and Margarita Majem Subject: PD-(L)1 inhibitors are part of the treatment strategy for non-small cell lung cancer (NSCLC) although its efficacy is limited to certain patients. Our study aimed to identify patients who might benefit from anti-PD-(L)1 inhibitors by analyzing the PD-L1 expression on circulating leukocytes and its evolution during treatment. One hundred thirteen NSCLC patients, according to their radiological response after 10 12 weeks of treatment, were classified into responders, stable, and progressive disease. Percentages of circulating PD-L1+ leukocytes, PD-L1+ platelets (PLTs), and leukocyte-PLT complexes were assessed using flow cytometry, and plasma concentrations of soluble immunomodulatory factors were quantified by ELISA. Responders exhibited significantly higher pre-treatment percentages of PD-L1+ neutrophils, PD-L1+ CD14+ cells, and PD-L1+ PLTs than progressors. The percentages of these populations decreased in responders post-treatment, contrasting with stables and progressors. PLTs notably contributed to PD-L1 expression in CD14+ cells and neutrophils. Plasma cytokine analysis revealed baseline differences only in IL-17 concentration among groups, whereas network analyses highlighted distinct association patterns between plasma molecules and PD-L1+ leukocytes after 10 12 weeks of treatment. Our findings suggest that pre-treatment assessment of circulating PD-L1+ neutrophils, PD-L1+ CD14+ cells, and PD-L1+ PLTs may be helpful in identifying NSCLC patients who are potential candidates for anti-PD-(L)1 therapy. 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