Flash Talk + Poster Presentation 37th Lorne Cancer Conference 2025

Identification of Multipotent Progenitor Cells in Oesophageal Submucosal Glands through Single-Cell Transcriptomics and Organoid Cultures: Implications for Barrett’s Metaplasia (#102)

Ibukun IA Adejumobi 1 2 , Elhadi Lich 1 2 , Jessica Tang 3 4 5 , Susanne Ramm 1 6 , Fernando Rossello 3 4 5 7 , Nicholas Clemons 1 2 , Wayne Phillips 1 2
  1. Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia
  2. Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne , Victoria, Australia
  3. Novo Nordisk Foundation Centre for Stem Cell Medicine (reNEW), Murdoch Children's Research Institute, Melbourne , Victoria
  4. Murdoch Children’s Research Institute, The Royal Children’s Hospital, Melbourne , Victoria , Australia
  5. Department of Clinical Pathology, University of Melbourne, Melbourne, Melbourne , Victoria , Australia
  6. Victoria Centre for Functional Genomics, Peter MacCallum Cancer Centre, Melbourne, Melbourne , Victoria , Australia
  7. Australian Regenerative Medicine Institute, Monash University, Melbourne , Victoria , Australia

Background: Barrett’s oesophagus is a premalignant condition where normal squamous epithelium is replaced by metaplastic columnar epithelium, increasing the risk of oesophageal adenocarcinoma. Gastroesophageal reflux disease (GERD) is a major risk factor for Barrett's oesophagus, but the cellular origins and mechanisms driving this metaplasia remain unclear. Oesophageal submucosal glands (SMGs) are hypothesized to harbour progenitor cells crucial for tissue regeneration and potentially implicated in metaplastic transformations associated with chronic GERD.

Aim: To identify and characterize multipotent progenitor cells within SMGs and investigate their potential role in Barrett’s metaplasia.

Methods: We employed single-cell RNA sequencing (scRNA-seq) and organoid cultures derived from porcine SMGs, validated as a model for human SMGs through multiplex immunohistochemistry and lectin staining. Organoid cultures were established to assess the differentiation potential of these cells. StemID and pseudotime trajectory analyses were utilized to identify progenitor populations and map their differentiation pathways.

Results: scRNA-seq of EpCAM⁺ epithelial cells identified distinct cellular subpopulations. We identified LGR6⁺ myoepithelial and basal duct cells within SMGs as multipotent progenitors capable of differentiating into both squamous and secretory lineages. Organoid cultures recapitulated native cellular diversity, exhibiting morphologies corresponding to specific cell types. Functional assays confirmed the multipotent capacity of these progenitors. Comparative analysis with published human SMG scRNA-seq data validated our findings, revealing conserved gene expression patterns and cellular hierarchies, including Barrett’s oesophagus–associated genes such as MUC5AC, TFF3, AGR2, and SOX9.

Conclusion: Our study highlights the critical role of LGR6⁺ multipotent progenitor cells within SMGs in maintaining oesophageal homeostasis and their potential contribution to metaplastic transformation under pathological conditions induced by GERD. Targeting these progenitors or their differentiation pathways may offer novel therapeutic strategies for preventing or reversing Barrett’s oesophagus.