Effect of Calcrl deletion on cytokine expression in renal monocytes and macrophages
Calcitonin gene-related peptide (CGRP) released from TRPV1+ sensory neurons protects against renal ischemia-reperfusion (I/R) injury by restraining myeloid-driven inflammation. CGRP signals through the CALCRL-RAMP1 receptor complex on monocytes/macrophages, but how this pathway shapes their transcriptional programs in vivo remains unclear. To address this, we performed bulk RNA sequencing of renal monocytes/macrophages sorted from Lyz2cre+/-; Calcrl fl/fl mice with myeloid-specific deletion of Calcrl and Calcrl fl/fl control mice 2 days after bilateral 35 minutes renal I/R injury. Cells were purified using a two-step strategy, consisting of magnetic enrichment of CD11b+ cells (MACS) followed by FACS sorting of CD11bhi Ly6Glo CD11clo monocytes/macrophages. and libraries were prepared using Smart-seq2 and sequenced on an Illumina NovaSeq platform. Comparative transcriptomic analysis revealed that loss of myeloid Calcrl amplifies pro-inflammatory and chemotactic gene expression, including TNF, Toll-like receptor, and NF-kappaB pathway components and key cytokines/chemokines (e.g., Tnf, Il1b, Cxcl1/2/10, Ccl2/3/4/5/7). These data define the CGRP-CALCRL axis as a critical regulator of monocyte/macrophage inflammatory programs in the kidney following ischemic injury.