10x Single-cell Sequencing for Smad3 wild-type and knockout neutrophils

ABSTRACT: The role of neutrophils in tumor is still controversial and largely unknown. We previously identified an anticancer tumor microenvironment from mice lacking Smad3, but the underlying mechanism is still largely unclear. Interestingly, depletion of neutrophils largely suppressed the anticancer phenotype of Smad3-KO mice against syngeneic murine Lewis lung carcinoma (LLC). Here, we discovered that Smad3 is essential for promoting N1/N2 polarization of the tumor_x0002_associated neutrophils (TANs) in lung carcinoma. We found that Smad3 deficiency dramatically increased N1 TANs associated with an anticancer phenotype in mice and detected a negative correlation between Smad3 activation and anticancer N1 associated with a higher mortality of NSCLC. By conducting a neutrophil-specific single-cell RNA-sequencing, we uncovered that Smad3 is essential for completing the N1/N2 polarization of TANs, whereas N1 will be the dominated phenotype in the Smad3-KO TME by analyzing the developmental pathways of TANs with pseudo-time at transcriptome level. Mechanistically, direct binding of Smad3 on the developmental genes in neutrophils under cancer condition was evidenced by ChIP-sequencing at genomic level. Importantly, both neutrophil-specific and pharmaceutical inhibition of Smad3 effectively enhanced the anticancer activity of human and murine TANs, thereby suppressing the progression of lung carcinoma in vivo. Thus, Smad3 may represent a precision therapeutic target for enhancing the anticancer activity of TANs via blocking N1/N2 polarization in NSCLC. in C57BL6 (wild-type) and Smad3-deficient (Smad3-/- ) mice and submitted for cell encapsulation and library construction by Chromium controller with 5’ expression kit (10x genomics)

ORGANISM(S): Mus musculus

PROVIDER: GSE186530 | GEO | 2023/03/16

REPOSITORIES: GEO