Replication Data for: Mycobacterium-based immunotherapy modulates microbiota in the orthotopic murine model of bladder cancer

Datasets for supplemental figures of binary survival data (Supplementary Figure 1) and body weight dataset (Supplementary Figure 2), for the study "Mycobacterium-based immunotherapy modulates microbiota in the orthotopic murine model of bladder cancer":

The once-held belief in the sterility of the urinary tract has been challenged by the discovery of a resident urinary microbiota. Some studies have observed differences in microbiota between healthy individuals and bladder cancer patients, although results show disparity, and a clear differentiated profile has not been proven. The bladder microbiota may influence tumor progression and response to therapies, including Mycobacterium bovis BCG intravesical instillations. The interference or synergy mechanisms between bladder and/or gut microbiota and BCG treatment remain unknown. This study evaluates for the first time the impact of intravesical instillations of BCG and Mycobacterium brumae, a non-pathogenic immunomodulatory species, on microbiota composition of bladder tissue and cecal stool in healthy and tumor-bearing mice.

Our results showed that mycobacterial instillations enhanced microbial richness but reduced Actinobacteria abundance across groups in the gut. At the family level, Muribaculaceae and Bacteroidaceae were significantly enriched in tumor-bearing mice, whereas BCG-treated animals showed lower Muribaculaceae and higher Rikenellaceae abundance. Regarding bladder microbiota, Staphylococcaceae dominated in M. brumae-treated healthy mice, while Enterococcaceae was abundant in all other groups except BCG-treated tumor-bearing mice. At genus level, Escherichia and Shigella were markedly reduced in mycobacteria-treated tumor-bearing mice, while Burkholderia was more prevalent in untreated bladders. Alpha diversity increased in cecal stool following M. brumae instillations, regardless of tumor status. In contrast, BCG reduced bladder microbiota richness in tumor-bearing mice. Beta diversity analyses revealed subtle, non-significant clustering among groups. These findings highlight how intravesical mycobacterial therapy modulates richness and diversity of microbiota composition in distinct anatomical sites, underlying the complex interplay between host, tumor, and immunotherapy.