About this item:

87 Views | 59 Downloads

Author Notes:

Sampath Prahalad, Email: sprahal@emory.edu

KI performed most of the single cell data analysis, with assistance from MD and VP. NT conceived the study, recruited patients, supervised the sample acquisition and phenotyping. LP recruited patients and organized participant data. NT and TG were responsible for specimen processing, cell preparation and TNF stimulation, with advice and support of AK, under the supervision of EG who also helped in study design. AK and DA were responsible for generation of single-cell gene expression libraries. SC helped with study design and interpretation of results. SP conceived the study, provided clinical oversight, and edited the manuscript. GG wrote the first draft of the manuscript and supervised the analyses. All authors read and approved the final manuscript.

In addition to the generous participation of all donors, we gratefully acknowledge discussions with James R. Rose while he was rotating in the Ghosn lab, Subra Kugathasan for critical input. We acknowledge Drs. Amit Thakral, Elaine Flanagan and Angela Taneja for referring subjects to the study. Savannah Washburn kindly assisted with the data deposition to GEO. We acknowledge the Molecular Evolution Core the Parker H. Petit Institute for Bioengineering and Bioscience at the Georgia Institute of Technology for the single-cell library sequencing; and Emory Integrated Genomics Core (EIGC), which is subsidized by the Emory University School of Medicine and is one of the Emory Integrated Core Facilities for performing QC on single-cell libraries.

Dr. Prahalad has served on a macrophage activation syndrome adjudication committee for Novartis, but this had no bearing on the work described in this manuscript. None of the other authors have any competing interests in the manuscript.

Kathleen J. Imbach and Nicole J. Treadway contributed equally to this work. Sampath Prahalad and Greg Gibson are shared senior authors.

Subjects:

Research Funding:

This work was supported by Marcus Foundation funds awarded to SP, and a Children’s Healthcare of Atlanta, Pediatric Research Alliance seed grant to EG, whose group is partially supported by start-up from the Lowance Center for Human Immunology at Emory University. KJI was a Beckman Foundation undergraduate research scholar, and NJT was partially supported by a Rheumatology Research Foundation medical and graduate student preceptorship award.

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Pediatrics
  • Rheumatology
  • JIA
  • Transcriptomics
  • Immunoprofiling
  • scRNAseq
  • BLOOD GENE-EXPRESSION
  • ONSET
  • IDENTIFICATION
  • HETEROGENEITY
  • ASSOCIATION
  • AUTOIMMUNE
  • THERAPY

Profiling the peripheral immune response to ex vivo TNF stimulation in untreated juvenile idiopathic arthritis using single cell RNA sequencing

Show all authors Show less authors

Tools:

Journal Title:

PEDIATRIC RHEUMATOLOGY

Volume:

Volume 21, Number 1

Publisher:

, Pages 17-17

Type of Work:

Article | Final Publisher PDF

Abstract:

Background: Juvenile Idiopathic Arthritis (JIA) is an autoimmune disease with a heterogenous clinical presentation and unpredictable response to available therapies. This personalized transcriptomics study sought proof-of-concept for single-cell RNA sequencing to characterize patient-specific immune profiles. Methods: Whole blood samples from six untreated children, newly diagnosed with JIA, and two healthy controls were cultured for 24 h with or without ex vivo TNF stimulation and subjected to scRNAseq to examine cellular populations and transcript expression in PBMCs. A novel analytical pipeline, scPool, was developed wherein cells are first pooled into pseudocells prior to expression analysis, facilitating variance partitioning of the effects of TNF stimulus, JIA disease status, and individual donor. Results: Seventeen robust immune cell-types were identified, the abundance of which was significantly affected by TNF stimulus, which resulted in notable elevation of memory CD8 + T-cells and NK56 cells, but down-regulation of naïve B-cell proportions. Memory CD8 + and CD4 + T-cells were also both reduced in the JIA cases relative to two controls. Significant differential expression responses to TNF stimulus were also characterized, with monocytes showing more transcriptional shifts than T-lymphocyte subsets, while the B-cell response was more limited. We also show that donor variability exceeds the small degree of possible intrinsic differentiation between JIA and control profiles. An incidental finding of interest was association of HLA-DQA2 and HLA-DRB5 expression with JIA status. Conclusions: These results support the development of personalized immune-profiling combined with ex-vivo immune stimulation for evaluation of patient-specific modes of immune cell activity in autoimmune rheumatic disease.

Copyright information:

© The Author(s) 2023

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/rdf).
Export to EndNote