Functional immune profiling in peripheral mononuclear cells to inform personalized approaches in patients with systemic lupus erythematosus - FiLup

Project title: Functional immune profiling in peripheral mononuclear cells to inform personalized approaches in patients with systemic lupus erythematosus

Acronym: FiLup

Competition: P1 – Development of the national R&D system - Subprogramme 1.1 – Human Resources, Research projects for stimulating young independent teams

Project code: PN-III-P1-1.1-TE-2021-1512

Project duration: 24 months (13.05.2022 – 12.05.2024)

Budget: 450.000,00 lei

Coordinator: “Iuliu Hațieganu” University of Medicine and Pharmacy, Cluj-Napoca – RO, Rheumatology and Medical Genetics

Project director: Assist. prof. Cristina Pamfil, MD, PhD

Abstract

Systemic lupus erythematosus (SLE) is the prototypic autoimmune disease with extreme patient variability, diminished health-related quality of life and high economic burden. Transcriptomic analysis differentiates patients with SLE from healthy individuals but fails to segregate patients with active SLE from patients in remission. The hypothesis of this proposal is that trained immunity takes place in cells of patients with SLE due to stimulation with IFN-I or other components. This leads to a persistently altered transcriptional program which contributes to inflammatory vulnerability (flares, atherosclerosis) in otherwise successfully treated patients. We will conduct experiments that characterize the cytokine production profiles of peripheral blood mononuclear cells in patients with SLE and correlate them to disease status and IFN signature. We will assess humoral factors present in plasma of patients and determine their potential to induce trained immunity via epigenetic modifications. The results of this project will fill the knowledge gap on the functional immunology of SLE and will provide an invaluable new layer of information linking upstream regulatory mechanisms to cytokine production in SLE. The impact consists in the fact that cytokines are effector molecules which are targetable with therapy. Comprehensive bio profiling of functional PBMC responses would contribute to both understanding of pathogenesis and identification of therapy targets in SLE.

Project objectives

The project will pursue two key objectives (KO) and answer the research questions listed below:

KO1. To assess the cytokine production capacity in PBMCs of patients with SLE and controls

  1. Is there an altered cytokine production capacity in patients with SLE? – compare cytokine responses in SLE patients versus controls
  2. Is this cytokine production pattern persistent despite successful therapy? – compare cytokine responses in SLE patients with active disease versus patients in remission
  3. What are the gene variants that influence cytokine production in SLE? – perform cytokineQTL analysis and determine SLE specific loci associated to cytokine production

KO2. To study the potential of humoral factors of SLE plasma to induce trained immunity

  1. Does IFN status correlate to cytokine production in SLE patients? - assess the IFN signature induced by SLE patients’ plasma and compare cytokine production based on high or low-IFN
  2. Does IFN-I induce trained immunity in primary PBMCs? – perform in vitro experiments inducing trained immunity using IFN-I
  3. Do other humoral components of SLE plasma lead to persistent effects on cytokine production? – perform in vitro experiments inducing trained immunity using media supplemented with 10% plasma of patients with SLE or controls
  4. Are these effects reversible upon epigenetic modulation? – experiments as above in the presence or absence of epigenetic modulators