Analysis of Lower-Limb Ulcers in Participants with Leprosy Sequelae Using Metabolomics and 16S Ribosomal DNA Sequencing

Abstract

Purpose

This study investigated microbiome and metabolome differences between ulcerated tissues and normal skin from the lower limbs of participants with leprosy.

Patients and Methods

Ulcerated tissues and surrounding normal skin were collected from the lower limbs of 28 participants with leprosy who had been cured. The 16S ribosomal DNA sequencing analysis of the samples was conducted with the Illumina NovaSeq platform to analyze the community structure and diversity of microorganisms on the skin surface, followed by non-targeted metabolomic analysis with LC-MS technology. Next, differential metabolites were statistically screened, followed by metabolic pathway analysis. The Spearman method was used to analyze the correlation between differential microbiota and differential metabolites.

Results

Compared to normal skin, ulcerated tissues showed a decrease in microbial α diversity (species richness, homogeneity, and sequencing depth), without significant differences (observed species, Chao1, Shannon, Simpson, and Pielou’s evenness index; P > 0.05). Conversely, Jaccard distance demonstrated that sample β-diversity exhibited a certain degree of clustering (P < 0.05), with significant differences between the two groups. The results of LEfSe analysis revealed that compared to the normal skin, the ulcerated tissues had significantly decreased microbial abundance of Flavobacteriaceae, Flavobacteriales, Lachnospiraceae, Lachnospirales, Enterobacterales, Acinetobacter, and Moraxellaceae, which might be associated with the ulcerative state. The Spearman correlation analysis suggested a strong correlation between skin metabolome and skin microbiome.

Conclusion

For participants with leprosy sequelae, skin microecology and metabolites are disturbed and species diversity and homogeneity are reduced in lower-limb ulcers, and the types of skin metabolites are dependent on the microbiota.

Keywords:

• leprosy
• lower-limb ulcers
• 16S rDNA sequencing
• metabolome
• skin microecology

Data Sharing Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Ethics Approval and Consent to Participate

This study was designed in accordance with the Declaration of Helsinki and approved by the ethics committee of Zhejiang Province Dermatology Hospital (approval number: 2022-004K). Informed consent was obtained from all participants involved in the study.

Acknowledgment

We would like to acknowledge the reviewers for their helpful comments on this paper.

Author Contributions

1. Made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas: Jian Wang, Ben Wang, Chao Liang, Caifei Jin, Huiliang Shen.

2. Have drafted or written, or substantially revised or critically reviewed the article: Jian Wang, Ben Wang, Chao Liang, Caifei Jin, Huiliang Shen.

3. Have agreed on the journal to which the article will be submitted: Jian Wang, Ben Wang, Chao Liang, Caifei Jin, Huiliang Shen.

4. Reviewed and agreed on all versions of the article before submission, during revision, the final version accepted for publication, and any significant changes introduced at the proofing stage: Jian Wang, Ben Wang, Chao Liang, Caifei Jin, Huiliang Shen.

5. Agree to take responsibility and be accountable for the contents of the article: Jian Wang, Ben Wang, Chao Liang, Caifei Jin, Huiliang Shen.

Disclosure

The authors report no conflicts of interest in this work.

Additional information

Funding

This work was supported by the Zhejiang Health Technology plan [grant numbers 2022KY732].