Glutamine supplementation as a novel metabolic therapeutic strategy for LIG3-dependent chronic intestinal pseudo-obstruction.

ABSTRACT

BACKGROUND AND AIMS: We recently identified a recessive syndrome due to LIG3 mutations in patients with chronic intestinal pseudo-obstruction, leukoencephalopathy and neurogenic bladder. LIG3 mutations affect mitochondrial DNA (mtDNA) maintenance, leading to defective energy production. We aimed at identifying altered molecular pathways and develop possible targeted treatments to revert / ameliorate the cellular energy impairment. METHODS: Whole transcriptome analysis was performed on patients' derived fibroblasts total RNA and controls. Mitochondrial function, mitophagy, L-Glutamine (L-Gln) supplementation effects were analyzed by live cell analysis, immunostaining and western blot. Patients were treated with Dipeptiven according to standard protocols. Patients' symptoms were analyzed by the Gastrointestinal Symptom Rating Scale questionnaire. RESULTS: We identified deregulated transcripts in mutant fibroblasts vs. controls, including overexpression of genes involved in extracellular matrix development and remodeling and mitochondrial functions. Gut biopsies of LIG3-mutant patients documented collagen and elastic fiber accumulation. Mutant fibroblasts exhibited impaired mitochondrial mitophagy indicative of dysfunctional turnover and altered Ca2+ homeostasis. L-Gln supplementation (6 mM), previously shown to increase mtDNA-defective cell survival, improved growth rate and ATP production in LIG3-mutant fibroblasts. These data led us to provide parenterally a dipeptide containing L-Gln to three siblings carrying biallelic LIG3 mutations. Compared to baseline, gastrointestinal and extra-gastrointestinal symptoms significantly improved after 8 months of treatment. CONCLUSIONS: LIG3 deficiency leads to mitochondrial dysfunction. High levels L-Gln supplementation was beneficial in LIG3-mutant cells and improved symptom severity without noticeable side effects. Our results provide a proof-of-concept to design ad hoc clinical trials with L-Gln in LIG3-mutant patients.