Additional file 2 of Beneficial effects of recombinant CER-001 high-density lipoprotein infusion in sepsis: results from a bench to bedside translational research project
Creators: name: Stasi, Alessandra
name: Fiorentino, Marco
name: Franzin, Rossana
name: Staffieri, Francesco
name: Carparelli, Sabrina
name: Losapio, Rosa
name: Crovace, Alberto
name: Lacitignola, Luca
name: Cimmarusti, Maria Teresa
name: Murgolo, Francesco
name: Stufano, Monica
name: Cafiero, Cesira
name: Castellano, Giuseppe
name: Sallustio, Fabio
name: Ferrari, Chiara
name: Ribezzi, Mario
name: Brienza, Nicola
name: Schirinzi, Annalisa
name: Di Serio, Francesca
name: Grasso, Salvatore
name: Pontrelli, Paola
name: Tupin, Cyrille
name: Barbaras, Ronald
name: Keyserling-Peyrottes, Constance
name: Crovace, Antonio
name: Gesualdo, Loreto
Publisher: name: figshare
Year: 2024
Subjects: subject: Medicine
subject: Pharmacology
subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 2: Figure S1. RACERS Trial design. Figure S2. In vitro effects of CER-001 on LPS-treated endothelial cells and PBMCs. A-B. Cultured endothelial cells were stimulated with LPS at 0.3 ug/ml and/or CER-001 at 50 and 500 ug/ml for 60. FACS (Fluorescence Activated Cell Sorting) showed a strong decrease of eNOS (phospho S1177) (p-ENOS) after 60’ of LPS stimulation compared to basal and VEGF as a positive control. CER-001 supplementation at 500 ug/ml, completely reversed LPS effects. A. Representative data from one out of a total of three experiments are shown. B. Histograms indicate p-ENOS expression levels. C-E. PBMC from 3 different healthy donors were stimulated with LPS at 0.3 ug/ml and/or CER-001 at 50 and 500 ug/ml for 24 h. C-D FACS showed a strong upregulation of CD14 surface expression 24 h following LPS stimulation. PBMCs treated with LPS and CER-001 in combination maintained CD14 expression at basal level. C. One representative of three independent experiments is shown. D. Histograms indicate CD14 expression levels. E. PBMC culture supernatants were analyzed by ELISA. After 24 h from LPS stimulation, PBMCs increased TNF-α synthesis. Stimulation of PBMCs with CER-001 at 50 and 500 ug/ml alone did not influence TNF-α production. The addition of CER-001, both at 50 and 500 ug/ml, in culture media of LPS-activated PBMCs reverted LPS effects. A-E Data are representative of three independent experiments. Data are shown as mean ± standard deviation (SD) and compared with the Student-t test. Figure S3. Effect of CER-001 on ApoA-I level, LPS removal and inflammatory response in the pilot study RACERS presented by individual treatment groups. A-I. Serum levels of ApoA-I, LPS, IL-6, IL-8, MCP1, TNF-α, sTREM-1, VCAM and ICAM were measured using ELISA kits. Data for panels A2-I are represented as changes from baseline (Day 1 pre-dose) values. Treatment differences were assessed by using a mixed model ANOVA for values on Days 3, 6 and 9 with a significance level of α = 0.05; significant pairwise comparisons are shown. No adjustments were made for multiple comparisons. Abbreviations: ApoA-I apolipoprotein A-I; ICAM endothelial intercellular adhesion molecule; IL-6 Interleukin 6; IL-8 Interleukin 8; IL-10 Interleukin 10; LPS lipopolysaccharides; MCP-1 Monocyte Chemoattractant Protein-1; TNF-α Tumor Necrosis factor α; sTREM-1 soluble Triggering receptor expressed on myeloid cells 1; VCAM vascular cell adhesion molecule. Figure S4. Effect of CER-001 on length of stay and need for organ support in ICU patients. A-B. Days in ICU presented as survival curve and as mean/SEM number of days. C. Daily average MAP. Data is presented as change from Baseline (24 h preceding first dose) and analyzed using a mixed model ANOVA for values on Days 1 through 9 with a significance level of α = 0.05; significant pairwise comparisons are shown. No adjustments were made for multiple comparisons. D. Number of days with vasopressor use. E. Number of days with mechanical ventilation use. F. Number of days on dialysis. G-H Days alive without organ support presented as survival curve and as mean/SEM number of days.