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  • Rectal organoid morphology analysis (ROMA) as a novel physiological assay for diagnostic classification in cystic fibrosis

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Cystic fibrosis
Original research
Rectal organoid morphology analysis (ROMA) as a novel physiological assay for diagnostic classification in cystic fibrosis
  1. Senne Cuyx1,2,
  2. Anabela Santo Ramalho1,
  3. Steffen Fieuws3,4,
  4. Nikky Corthout5,6,
  5. Marijke Proesmans1,2,
  6. Mieke Boon1,2,
  7. Kaline Arnauts7,8,
  8. Marianne S Carlon9,10,
  9. Sebastian Munck5,6,
  10. Lieven Dupont10,11,
  11. Kris De Boeck1,2,
  12. François Vermeulen1,2
  13. on behalf of the Belgian Organoid Project
    1. 1 Department of Development and Regeneration, Woman and Child Unit, CF Research Lab, KU Leuven, Leuven, Belgium
    2. 2 Department of Pediatrics, Pediatric Pulmonology, University Hospitals Leuven, Leuven, Belgium
    3. 3 Interuniversity Center for Biostatistics and Statistical Bioinformatics, KU Leuven, Leuven, Belgium
    4. 4 Interuniversity Center for Biostatistics and Statistical Bioinformatics, Hasselt University, Hasselt, Belgium
    5. 5 VIB Bio Imaging Core and VIB-KU Leuven Center for Brain & Disease Research, KU Leuven, Leuven, Belgium
    6. 6 Department of Neuroscience, KU Leuven, Leuven, Belgium
    7. 7 Department of Chronic Diseases, Metabolism and Ageing (CHROMETA), Translational Research Center for Gastrointestinal Disorders (TARGID), KU Leuven, Leuven, Belgium
    8. 8 Department of Gastroenterology and Hepatology, University Hospitals Leuven, Leuven, Belgium
    9. 9 Center for Molecular Medicine, KU Leuven, Leuven, Belgium
    10. 10 Department of Chronic Diseases and Metabolism, Laboratory of Respiratory Diseases and Thoracic Surgery (BREATHE), KU Leuven, Leuven, Belgium
    11. 11 Department of Respiratory Diseases, University Hospitals Leuven, Leuven, Belgium
    1. Correspondence to Dr François Vermeulen, Department of Development and Regeneration, Woman and Child Unit, CF Research Lab, KU Leuven, Leuven 3000, Belgium; francois.vermeulen{at}uzleuven.be

    Abstract

    Background Diagnosing cystic fibrosis (CF) is not always straightforward, in particular when sweat chloride concentration (SCC) is intermediate and <2 CF-causing CFTR variants are identified. The physiological CFTR assays proposed in the guidelines, nasal potential difference and intestinal current measurement, are not readily available nor feasible at all ages. Rectal organoid morphology analysis (ROMA) was previously shown to discriminate between organoids from subjects with and without CF based on a distinct phenotypical difference: compared with non-CF organoids, CF organoids have an irregular shape and lack a visible lumen. The current study serves to further explore the role of ROMA when a CF diagnosis is inconclusive.

    Methods Organoid morphology was analysed using the previously established ROMA protocol. Two indices were calculated: the circularity index to quantify the roundness of organoids and the intensity ratio as a measure of the presence of a central lumen.

    Results Rectal organoids from 116 subjects were cultured and analysed together with the 189 subjects from the previous study. ROMA almost completely discriminated between CF and non-CF. ROMA indices correlated with SCC, pancreatic status and genetics, demonstrating convergent validity. For cases with an inconclusive diagnosis according to current guidelines, ROMA provided additional diagnostic information, with a diagnostic ROMA classification for 18 of 24 (75%).

    Discussion ROMA provides additional information to support a CF diagnosis when SCC and genetics are insufficient for diagnostic classification. ROMA is standardised and can be centralised, allowing future inclusion in the diagnostic work-up as first-choice physiological assay in case of an unclear diagnosis.

    • Cystic Fibrosis

    Data availability statement

    All data relevant to the study are included in the article or uploaded as supplemental information.

    https://doi.org/10.1136/thorax-2023-220964

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      Data availability statement

      All data relevant to the study are included in the article or uploaded as supplemental information.

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      Footnotes

      • X @Carlon lab @CarlonMarianne

      • Collaborators Belgian Organoid Project: Hedwige Boboli (CHR Citadelle, Liège, Belgium), Linda Boulanger (University Hospital Leuven, Belgium), Georges Casimir (HUDERF, Brussels, Belgium), Benedicte Demeyere (Ghent University Hospital, Belgium), Elke De Wachter (University Hospital Brussels, Belgium), Danny De Looze (Ghent University Hospital, Belgium), Anja Delporte (Ghent University Hospital, Belgium), Isabelle Etienne (CHU Erasme, Brussels, Belgium), Laurence Hanssens (HUDERF, Brussels), Christiane Knoop (CHU Erasme, Brussels, Belgium), Monique Lequesne (University Hospital Antwerp, Belgium), Vicky Nowé (GZA St Vincentius Hospital Antwerp), Ann Raman (Ghent University Hospital, Belgium), Dirk Staessen (GZA St Vincentius Hospital Antwerp), Stephanie Van Biervliet (Ghent University Hospital, Belgium), Eva Van Braeckel (Ghent University Hospital, Belgium), Kim Van Hoorenbeeck (University Hospital Antwerp, Belgium), Eef Vanderhelst (University Hospital Brussels, Belgium), Stijn Verhulst (University Hospital Antwerp, Belgium), Stefanie Vincken (University Hospital Brussels, Belgium).

      • Contributors SC—conceptualisation, methodology, software, validation, formal analysis, investigation, resources, data curation, writing (original draft), writing (review and editing), visualisation, funding acquisition, project administration. ASR—conceptualisation, methodology, software, validation, formal analysis, investigation, resources, data curation, writing (original draft), writing (review and editing), visualisation, funding acquisition, project administration, supervision. SF—conceptualisation, methodology, software, formal analysis, investigation, data curation, writing (original draft), writing (review and editing), visualisation. NC—conceptualisation, methodology, software, formal analysis, investigation, resources, visualisation. MP—resources, writing (review and editing), funding acquisition, project administration. MB—resources, writing (review and editing), funding acquisition, project administration. KA—resources, data curation, writing (review and editing). MSC—resources, writing (review and editing), funding acquisition. SM—conceptualisation, methodology, software, resources, visualisation. LD—resources, writing (review and editing), funding acquisition, project administration. CADB—conceptualisation, methodology, validation, formal analysis, investigation, resources, data curation, writing (review and editing), funding acquisition, project administration, supervision. FV—conceptualisation, methodology, software, validation, formal analysis, investigation, resources, data curation, writing (original draft), writing (review and editing), visualisation, funding acquisition, project administration, supervision, guarantor.

      • Funding This work was supported by a grant from the Belgian cystic fibrosis patient association 'Association Muco/Mucovereniging' to the Belgian Organoid Project and the CF research centre at KU Leuven, by the Research Grant of the Belgian Society of Paediatrics (BVK-SBP 2019), by a grant from the UZ Leuven Fund for Translational Biomedical Research and by the Klosterfrau Group Award for research in Paediatric Pulmonology awarded by the Klosterfrau Healthcare Group (no award/grant numbers).

      • Competing interests None declared.

      • Provenance and peer review Not commissioned; externally peer reviewed.

      • Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.

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