Case report: Molecular characterisation of adipose-tissue derived cells from a patient with ROHHAD syndrome

Kalina M Biernacka; Dinesh Giri; Katherine Hawton; Francisca Segers; Claire M Perks; Julian P Hamilton-Shield

doi:10.3389/fped.2023.1128216

Case report: Molecular characterisation of adipose-tissue derived cells from a patient with ROHHAD syndrome

Front Pediatr. 2023 Jun 30:11:1128216. doi: 10.3389/fped.2023.1128216. eCollection 2023.

Authors

Kalina M Biernacka¹, Dinesh Giri², Katherine Hawton², Francisca Segers³, Claire M Perks^#¹, Julian P Hamilton-Shield^#⁴

Affiliations

¹ Cancer Endocrinology Group, Bristol Medical School, Translational Health Sciences, Southmead Hospital, Bristol, United Kingdom.
² Department of Paediatric Endocrinology and Diabetes, Bristol Royal Hospital for Children, Bristol, United Kingdom.
³ School of Biological Sciences, University of Bristol, Bristol, United Kingdom.
⁴ Department of Translational Health Sciences, Nutrition Theme, NIHR Bristol Biomedical Research Centre, Bristol Medical School, University of Bristol, UBHT Education Centre, Bristol, United Kingdom.

^# Contributed equally.

Abstract

There have been over 100 cases of Rapid-onset obesity with hypothalamic dysfunction, hypoventilation, and autonomic dysregulation (ROHHAD) syndrome reported, but there is currently no curative treatment for children with this condition. We aimed to better characterise adipose cells from a child with ROHHAD syndrome. We isolated pre-adipocytes from a 4 year-old female patient with ROHHAD syndrome and assessed proliferation rate of these cells. We evaluated levels of DLP-Pref-1(pre-adipocyte marker) using western blotting, and concentrations of interleukin-6(IL-6) using ELISA. We performed next-generation sequencing (NGS) and bioinformatic analyses on these cells compared to tissue from an age/sex-matched control. The two most up-/down-regulated genes were validated using QPCR. We successfully isolated pre-adipocytes from a fat biopsy, by confirming the presence of Pref-1 and differentiated them to mature adipocytes. Interleukin 6, (Il-6) levels were 5.6-fold higher in ROHHAD cells compared to a control age/sex-matched biopsy. NGS revealed 25,703 differentially expressed genes (DEGs) from ROHHAD cells vs. control of which 2,237 genes were significantly altered. The 20 most significantly up/down-regulated genes were selected for discussion. This paper describes the first transcriptomic analysis of adipose cells from a child with ROHHAD vs. normal control adipose tissue as a first step in identifying targetable pathways/mechanisms underlying this condition with novel therapeutic interventions.

Keywords: RNA sequencing; ROHHAD syndrome; adipocytes; immune modulation; interleukin-6 and cancer.

Publication types

Case Reports

Grants and funding

This research was funded by NIHR Applied Research Collaboration (ARC) West and NIHR Bristol Biomedical Research Centre (BRC), Nutrition theme. We also wish to acknowledge the family of the patient who provided consent to allow us a biopsy sample and produce this paper. The work was also supported by Cancer Research UK: The Integrative Cancer Epidemiology Programme for improved causal evidence and enhanced prediction of cancer risk and survival: C18281/A29019.