Deleting an Nr4a1 Super-Enhancer Subdomain Ablates Ly6C^low Monocytes while Preserving Macrophage Gene Function

Highlights

• The Nr4a1 enhancer domain regulates Ly6C^low monocyte development

• E2 domain-deficient resident tissue macrophages show wild-type Nr4a1 expression

• E2 domain-deficient macrophages respond normally to LPS stimulation

• KLF2 acts via E2 to regulate Nr4a1 in Ly6C^low monocyte development

Summary

Mononuclear phagocytes are a heterogeneous family that occupy all tissues and assume numerous roles to support tissue function and systemic homeostasis. Our ability to dissect the roles of individual subsets is limited by a lack of technologies that ablate gene function within specific mononuclear phagocyte sub-populations. Using Nr4a1-dependent Ly6C^low monocytes, we present a proof-of-principle approach that addresses these limitations. Combining ChIP-seq and molecular approaches we identified a single, conserved, sub-domain within the Nr4a1 enhancer that was essential for Ly6C^low monocyte development. Mice lacking this enhancer lacked Ly6C^low monocytes but retained Nr4a1 gene expression in macrophages during steady state and in response to LPS. Because Nr4a1 regulates inflammatory gene expression and differentiation of Ly6C^low monocytes, decoupling these processes allows Ly6C^low monocytes to be studied independently.