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Hedgehog blockade remodels the gut microbiota and the intestinal effector CD8+ T cells in a mouse model of mammary carcinoma


Given the gut microbiome’s rise as a potential frontier in cancer pathogenesis and therapy, leveraging microbial analyses in the study of breast tumor progression and treatment could unveil novel interactions between commensal bacteria and disease outcomes. In breast cancer, the Hedgehog (Hh) signaling pathway is a potential target for treatment due to its aberrant activation leading to poorer prognoses and drug resistance. There are limited studies that have investigated the influences of orally administered cancer therapeutics, such as Vismodegib (a pharmacological, clinically used Hh inhibitor) on the gut microbiota. Using a 4T1 mammary carcinoma mouse model and 16 S rRNA sequencing, we longitudinally mapped alterations in immunomodulating gut microbes during mammary tumor development. Next, we identified changes in the abundance of commensal microbiota in response to Vismodegib treatment of 4T1 mammary tumor-bearing mice. In addition to remodeling gut microbiota, Vismodegib treatment elicited an increase in proliferative CD8+ T cells in the colonic immune network, without any remarkable gastrointestinal-associated side effects. To our knowledge, this is the first study to assess longitudinal changes in the gut microbiome during mammary tumor development and progression. Our study also pioneers an investigation of the dynamic effects of an orally delivered Hh inhibitor on the gut microbiome and the gut-associated immune-regulatory adaptive effector CD8+ T cells. These findings inform future comprehensive studies on the consortium of altered microbes that can impact potential systemic immunomodulatory roles of Vismodegib.

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Fig. 1: Progression of 4T1 mammary tumor development and progression induces changes within the fecal microbiome.
Fig. 2: Hh inhibition impacts the gut bacterial abundance in the 4T1 immunocompetent model of mouse mammary carcinoma.
Fig. 3: Hh inhibition alters the α and β diversity of the fecal and cecal microbiota.
Fig. 4: Hh blockade progressively modifies the gut microbiota in mammary tumor-bearing mice.
Fig. 5: Smo-i treatment augments the proliferation of CD8+ T cells in the colon and the mesenteric lymph nodes.
Fig. 6: The oral administration of Smo-i does not induce colitis in 4T1 tumor-bearing mice.

Data availability

The datasets used and/or analyzed in this study are available from the corresponding author upon reasonable request.


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The authors acknowledge funding from the following sources: Department of Defense (W81XWH-14-1-0516, W81XWH-18-1-0036, W81XWH-10-1-0755) and O’Neal Invests awarded to L.A.S. The work is supported in part by BXAl3374 (VA) and CA194048 (NCI/NIH) to R.S.S. The authors acknowledge funding provided by T32AI007051-42 (awarded to D.C.H.) and 2T32GM8361-29 (awarded to C.A.S.). The authors thank Sarah Kammerud and Brandon Metge for editorial suggestions. The authors would like to thank the UAB Comprehensive Flow Cytometry Core supported by NIH Grants P30 AR048311 and P30 AI027667 and the UAB Comprehensive Cancer Center’s Preclinical Imaging Shared Facility supported by NIH Grants P30 CA013148 and 1S1 0OD021697.

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All authors have read and approved the manuscript. D.C.H., A.H., C.A.S., C.L.M., P.A.M., and L.A.S. contributed to experimental design. D.C.H., A.H., C.A.S., C.L.M., P.A.M. were responsible for acquisition of data. D.C.H., C.A.S., D.C., C.L.M., P.A.M., G.L., B.C.M., R.S.S., and L.A.S. performed analysis and interpretation of data. C.A.S., D.C.H., A.H., R.S.S, D.C., C.L.M., P.A.S., G.L., B.C.M., and L.A.S. were involved in writing and revision of the manuscript.

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Correspondence to Lalita A. Shevde.

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Hinshaw, D.C., Swain, C.A., Chen, D. et al. Hedgehog blockade remodels the gut microbiota and the intestinal effector CD8+ T cells in a mouse model of mammary carcinoma. Lab Invest (2022).

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