直肠类器官形态分析（ROMA）作为结缔组织纤维化（cystic fibrosis）诊断分类的新型生理检测方法

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

THORAX

影响因子:7.7

分区:医学1区 Top / 呼吸系统2区

发表日期:2024 Aug 19

作者: Senne Cuyx, Anabela Santo Ramalho, Steffen Fieuws, Nikky Corthout, Marijke Proesmans, Mieke Boon, Kaline Arnauts, Marianne S Carlon, Sebastian Munck, Lieven Dupont, Kris De Boeck, François Vermeulen,

DOI: 10.1136/thorax-2023-220964

摘要

在诊断囊性纤维化（CF）时，尤其是当汗液氯浓度（SCC）值处于中间范围且未检测到2个致病性CFTR变异时，诊断并不总是明确的。指南中提出的生理CFTR检测方法——鼻潜能差（nasal potential difference）和肠道电流测定（intestinal current measurement）在所有年龄段都难以获得或操作不便。此前研究显示，直肠类器官形态分析（ROMA）可以根据明显的表型差异，区分CF患者和非CF患者：与非CF类器官相比，CF类器官形状不规则，缺乏可见的腔道。本文旨在进一步探讨在CF诊断不明确时，ROMA的作用。采用既定的ROMA方案对类器官形态进行分析，计算两个指数：圆形指数（circulality index）衡量类器官的圆度，以及强度比（intensity ratio）反映中心腔道的存在。分析了来自116名受试者的直肠类器官，并结合之前研究的189名受试者的数据。结果显示，ROMA几乎完全区分CF与非CF，指数与SCC、胰腺状态和遗传学密切相关，验证了其收敛效度。在当前指南下诊断尚无结论的病例中，ROMA提供了额外的诊断信息，18例（75%）获得了明确的诊断分类。ROMA可作为支持CF诊断的补充手段，尤其在SCC和遗传学难以明确诊断时。该方法已标准化，可集中管理，未来可作为疑难诊断的首选生理检测方法纳入临床诊断流程。

Abstract

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.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.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%).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.