Inositol hexakisphosphate kinases 1 and 2 (IP6K1/2) are involved in renal control of phosphate homeostasis

Abstract

Phosphate (Pi) is an indispensable component of living organisms. In the body, Pi homeostasis is tightly regulated by four main tissues: intestine, kidney, bone and parathyroid glands. There are multiple complex mechanisms underlying the regulation of serum Pi including hormones and transporters. Although plenty of studies have analysed Pi regulation, the mechanism with which Pi levels are sensed by mammalian cells remains elusive. Molecular mechanisms of Pi-sensing have been elucidated in detail in yeast and bacteria. However, most of the Pi-sensing proteins identified in yeast and bacteria are encoded by genes that are not conserved in mammals. As a major exception, inositol polyphosphates—which respond to changes in ambient Pi in yeast and, in turn, regulate the expression of Pi transporters—are also found in mammals. Here, we addressed the role of inositol polyphosphates and their biogenic enzymes, the IP6Ks (especially IP6K1 and 2), in cellular Pi sensing in mammals. Human and mouse kidneys express the IP6K1 and IP6K2 isoforms. Likewise, Opossum Kidney (OK) cells, a model for renal proximal tubules express IP6K1 and 2. Pharmacological inhibition of IP6K1/2 or CRISPR/Cas9 mediated knockout in OK cells downregulates Pi transporter mRNA, particularly NaPi-IIa/c. Ip6k1-/-2-/- KO cells also exhibited decreased Pi uptake (by 70%) and an abrogated adaptation to ambient Pi. To study the role of IP6K1 and 2 in vivo, we generated renal tubule-specific inducible ip6k1-/-2-/- KO mice: the mice had strongly reduced expression of renal Pi transporters, at the protein and mRNA levels, and lower serum Pi (by 20%) together with the transcriptional downregulation of many other transporters. Serum intact fibroblast growth factor 23—a principal phosphaturic hormone—was also substantially reduced (10-fold) in these mice. Our data indicate that the deletion of IP6K1 and 2 decreases transcription of renal Pi transporters both in vivo and in vitro, and that Ip6k1-/-2-/- cells are unable to adapt to ambient Pi. Therefore, IP6K1 and 2 play a critical role in renal Pi metabolism and, potentially, sensing.