GSE117704
mRNA- and RISC-sequencing of miR-106b-transfected juxtaglomerular (JG) cells
Chronic inflammation is known to contribute to the development of hypertension, but while the role of lymphocytes is well-established, the myeloid lineage, particularly as a possible mediator of renin-mediated hypertension, has not been studied. Vitamin D deficiency has pro-inflammatory consequences in monocytes, and it is linked to renin-mediated hypertension in mice. This prompted us to test the hypothesis that conditional knockout of the vitamin D receptor (VDR) in macrophages (KODMAC) would promote renin-dependent hypertension. We found that lack of macrophage vdr signaling was sufficient to induce high blood pressure (BP) in mice. Recipients of bone marrow from KODMAC mice experienced increased BP via activation of the renal renin-angiotensin system. Conversely BM transplants from mice with intact vdr signaling reduced BP in KODMAC mice by decreasing plasma renin and increasing urinary Na excretion. Deletion of myeloid vdr promotes vascular macrophage infiltration, increasing ROS-mediated scavenging of NO and reducing renal perfusion, resulting in increased plasma renin. Interestingly, deletion of macrophage vdr promotes their infiltration into the JG cell apparatus, regulating JG cell renin production by an unexpected communication between macrophages and JG cells through miR-106b-5p. Induction of macrophage endoplasmic reticulum stress phospho-PERK and CHOP pathways increased miR-106b-5p secretion, the uptake of which into JG cells induced renin production and release by repression of E2f1 and Pde3b transcription factors, constituting a novel mechanism by which inflammation causes hypertension.  Overall design: Cultured JG cells engineered to express YFP under the control of the renin promoter (R A Gomez) were transfected with a control (non-mammalian) microRNA precursor or miR-106b.  48 h after transfection, RNA was prepared from 4 replicate control and 4 replicate miR-106b cell cultures and mRNA (polyA-mRNA) was extracted.  In parallel, cell pellets were obtained from another 4 replicate control and miR-106b cell cultures and Ago2 was immunoprecipitated followed by RNA extraction.  Messenger RNA and Ago2-bound RNA were converted to Illumina RNA-sequencing libraries.