Rectal Organoid Morphology Analysis (ROMA): A Diagnostic Assay in Cystic Fibrosis

Overview

This protocol describes rectal organoid morphology analysis (ROMA), a novel diagnostic assay for cystic fibrosis (CF). ROMA can effectively discriminate between organoids from subjects with and without CF, particularly when specific genetic mutations are present.

Key Study Components

Area of Science

• Neuroscience
• Biology
• Genetics

Background

• Cystic fibrosis is a genetic disorder affecting the respiratory and digestive systems.
• CFTR function is critical for proper organoid morphology.
• ROMA utilizes morphological characteristics to assess CFTR function.
• The analysis is semi-automated and can be standardized across laboratories.

Purpose of Study

• To develop a reliable diagnostic tool for cystic fibrosis.
• To measure CFTR function through organoid morphology.
• To provide a method that can be used across all ages with low complication rates.

Methods Used

• Organoids are stained with calcium green one day after plating.
• The analysis involves measuring roundness (circularity index) and lumen presence (intensity ratio).
• Biopsies can be sent to a central laboratory for analysis.
• ROMA can also assess CFTR modulatory efficacy.

Main Results

• Analysis of 189 subjects showed perfect discrimination between CF and non-CF.
• ROMA is effective even with less than two disease-causing CFTR mutations.
• It can be performed at all ages with low complication rates.
• ROMA helps measure the degree of restoration of CFTR function.

Conclusions

• ROMA is a promising tool for diagnosing cystic fibrosis.
• The method provides a standardized approach to assess CFTR function.
• Further research could enhance its application in clinical settings.

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