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Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
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Alterations of the gut ecological and functional microenvironment in different stages of multiple sclerosis

Daiki Takewaki1,2,3, Wataru Suda4, Wakiro Sato1,2

  • 1Department of Immunology, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Kodaira, 187-8502 Tokyo, Japan.

Proceedings of the National Academy of Sciences of the United States of America
|August 26, 2020
PubMed

Insights

Gut microbiome alterations are linked to multiple sclerosis (MS) progression. Relapsing-remitting MS (RRMS) shows reduced short-chain fatty acid (SCFA) production, while secondary progressive MS (SPMS) exhibits increased DNA oxidation markers.

Area of Science:

  • Neuroimmunology
  • Microbiome Research
  • Metabolomics

Background:

  • Multiple sclerosis (MS) is a central nervous system autoimmune disease, often progressing from relapsing-remitting (RRMS) to secondary progressive (SPMS) forms.
  • SPMS is characterized by increasing disability and reduced treatment efficacy, suggesting underlying disease mechanism changes.
  • The gut microbiome's role in MS pathogenesis and progression remains incompletely understood.

Purpose of the Study:

  • To investigate microbial and functional differences in the gut microbiomes across different stages of MS, including RRMS and SPMS, compared to healthy controls (HCs).
  • To correlate these microbiome changes with clinical severity and metabolic profiles.

Main Methods:

  • Comparative analysis of gut microbiomes using 16S rRNA gene sequencing and whole metagenomic sequencing on fecal samples from RRMS, SPMS, and HCs.
  • Fecal metabolite analysis, including short-chain fatty acids (SCFAs) and sulfur metabolites.
  • Correlation of microbial and functional data with clinical severity.

Main Results:

  • Significant alterations in bacterial species abundance were observed in MS patients compared to HCs, notably a reduction in SCFA-producing bacteria.
  • RRMS patients showed reduced biosynthesis of butyrate and propionate.
  • While bacterial composition differences between RRMS and SPMS were limited, metagenomic functional data revealed increased microbial genes for DNA mismatch repair in SPMS. Sulfur metabolomics indicated an increased ratio of cysteine persulfide to cysteine in SPMS, suggesting potential gut oxidative stress.

Conclusions:

  • Gut ecological and functional microenvironments are significantly altered in different stages of MS.
  • Reduced SCFA biosynthesis is characteristic of RRMS, while SPMS is associated with elevated oxidative levels in the gut.
  • These findings highlight distinct gut microbiome signatures associated with MS progression, potentially impacting disease mechanisms.
Keywords:
oxidative stressrelapsing remitting multiple sclerosissecondary progressive multiple sclerosissulfur metabolomicswhole metagenomic sequencing

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