Fecal microbiota transplantation enhances cell therapy in a rat model of hypoganglionosis by SCFA‐induced MEK1/2 signaling pathway

Hirschsprung disease (HSCR), one of several neurocristopathies in children, is characterized by nerve loss in the large intestine and is mainly treated by surgery, which causes severe complications. Enteric neural crest‐derived cell (ENCC) transplantation is a potential therapeutic strategy; however, so far with poor efficacy. Here, we assessed whether and how fecal microbiota transplantation (FMT) could improve ENCC transplantation in a rat model of hypoganglionosis; a condition similar to HSCR, with less intestinal innervation. We found that the hypoganglionosis intestinal microenvironment negatively influenced the ENCC functional phenotype in vitro and in vivo . Combining 16S rDNA sequencing and targeted mass spectrometry revealed microbial dysbiosis and reduced short‐chain fatty acid (SCFA) production in the hypoganglionic gut. FMT increased the abundance of Bacteroides and Clostridium , SCFA production, and improved outcomes following ENCC transplantation. SCFAs alone stimulated ENCC proliferation, migration, and supported ENCC transplantation. Transcriptome‐wide mRNA sequencing identified MAPK signaling as the top differentially regulated pathway in response to SCFA exposure, and inhibition of MEK1/2 signaling abrogated the SCFA‐mediated effects on ENCC. This study demonstrates that FMT improves cell therapy for hypoganglionosis via short‐chain fatty acid metabolism‐induced MEK1/2 signaling. Synopsis Fecal microbiota transplantation synergy with enteric neural crest‐derived cell transplantation is tested in a rat model of hypoganglianosis to provide a non‐surgical treatment option of neurocristopathies including Hirschsprung disease. The intestinal microenvironment in a rat model of hypoganglionosis negatively influences enteric neural crest‐derived cell (ENCC) proliferation and migration Fecal microbiota transplantation (FMT) relieves hypoganglionosis symptoms and promotes ENCC transplantation in vitro and vivo. Some genera, particularly Bacteroides and Clostridium , increased after FMT, which coincided with short‐chain fatty acid (SCFA) production. SCFA exposure stimulated ENCC proliferation, migration, and supported ENCC transplantation in the treatment of hypoganglionosis via the MEK1/2 signaling pathway. Graphical Abstract Fecal microbiota transplantation enhances enteric neural crest‐derived cell transplantation in a rat model of hypoganglionosis supporting a non‐surgical treatment option of neurocristopathies including Hirschsprung disease.