OC 19.3 - The endothelial inflammatory repertoire: a multi-omics delineation of cytokine-induced endothelial inflammation states

Sunday, July 10, 2022

11:15 AM – 11:30 AM

Location: ExCel Center, ICC Capital Suite Room 17

Presentation Format: In-Person

Presenting Author(s)

Stijn Groten, MSc

Phd student
Department of Molecular Hematology, Sanquin Research, 1066 CX Amsterdam, the Netherlands
Amsterdam, Noord-Holland, Netherlands

Co-Author(s)

ES

Eva Smit

PhD student
Sanquin Research
Amsterdam, Noord-Holland, Netherlands
EJ

Esmee Janssen

PhD student
Sanquin Research
Amsterdam, Noord-Holland, Netherlands
Bv

Bart van den Eshof

PhD student
Sanquin Research
Amsterdam, Noord-Holland, Netherlands
FV

Floris Van Alphen

Technician
Department of Molecular Hematology, Sanquin Research, 1066 CX Amsterdam, the Netherlands
Amsterdam, Noord-Holland, Netherlands
Cv

Carmen van der Zwaan

Senior Technician
Sanquin Research
Amsterdam, Noord-Holland, Netherlands
Am

Alexander meijer

Head of department / Professor
Sanquin Research / Utrecht University
Amsterdam, Noord-Holland, Netherlands
AH

Arie Hoogendijk

PI
Sanquin Research
Amsterdam, Noord-Holland, Netherlands
Mv

Maartje van den Biggelaar, PhD

PI
Department of Molecular Hematology, Sanquin Research, 1066 CX Amsterdam, the Netherlands
Amsterdam, Noord-Holland, Netherlands

Background: Hemostasis and inflammation are tightly intertwined processes in which vascular endothelial cells (ECs) form a dynamic interface between blood and tissue mediating critical steps in maintaining both anti- and pro- coagulation and inflammatory states. Detailed insight on the mechanistic processes that trigger endothelial inflammation could contribute to the design of intervention strategies at the crossroads between vascular inflammation and hemostasis.

Aims: Understanding the diversity of endothelial inflammatory responses.

Methods: To generate an overview of ECs responses, we evaluated changes in the proteome of blood derived ECs stimulated to a panel of 92 cytokines separately and in combination. We then performed a time-resolved multi-omics analysis on ECs exposed to TNFα and IFNγ, integrating transcriptome, whole (phospho-) proteome, and secretome (Fig.1).

Results: Inflammatory cytokines TNFα and IFNγ induced the highest number of significant proteomic events and exhibited distinct proteomic profiles. Our multi-omics integration allowed for in-depth mapping of endothelial inflammatory responses and highlighted distinct signatures as well as a drastically increased response through TNFα and IFNγ co-stimulation on all omics levels. Regulatory hubs correlated to TNFα activation of the NFKB pathway and IFNγ signaling through the JAK/STAT pathway (Fig.2). IFNγ also strongly upregulated MHCI and MHCII histocompatibility complexes and complement factors. Interestingly, several mRNAs, such as RelA, JAK3 and PECAM1 were synergistically regulated through combined TNFα and IFNγ stimulation. Moreover, ECs secreted a select subset of cytokines per inflammatory stimuli, which were synergistically increased through co-stimulation, including CCL5, CCL8, CXCL9 and IL6.

Conclusion(s): In conclusion, our integrated analysis reveals 1) an in-depth molecular mapping of endothelial inflammation on multiple omics levels, 2) the presence of distinct endothelial inflammatory states and 3) a synergistic endothelial response through combined TNFα and IFNγ stimulation. This study therefore supports the emerging role of ECs as active players in the progression of inflammation.