TmP/GFR = maximum reabsorption of PO4 per unit volume of GFR FEPO4 = UPO4 /PPO4 / Ucreat /Pcreat 1 - FEPO4 = TRP Assuming PPO4 = [PO4]GFR TmPO4/GFR = TRP X PPO4 • Tubular reabsoption of phosphate (TRP) • Tubular maximal reabsoption rate of phosphate to GFR (TmP/GFR) • TRP =99.9% • TmP/GFR = TRP x Plasma Phos = 0.999 x 0.93 mmol/L (converted from 2.9 mg/dL) = 0.93 mmol/L Regulation of Phosphate Transport in Proximal Tubule Alexander Usorov, MD 11/25/08 Overview • Role of phosphorus • Proximal nephron transport mechanism • Sodium-hydrogen exchanger regulatory factor-1 (NHERF1) • Fibroblast Growth Factor (FGF 23) Importance of Phosphorus • Component of hydroxyapatite, which is the major component of bone mineral • Present in nucleic acids, bioactive signaling proteins, phosophorylated enzymes, and cell membranes • Deficiency in phosphorus leads to – – – – Impaired bone mineralization (osteomalacia or rickets) Abnormal RBC, WBC, Plt fxn Impaired cell membrane integrity (rhabdo) Impaired cardiac output Phosphorus Metabolism 1500mg 1100mg 200mg 200mg Phosphorus reabsorption • Up to 95% occurs in the proximal tubule • Filtered phosphate moves from lumen to cells via Na-phosphate cotransporters located in the luminal membrane • Different types of Na/Pi cotransporters – NaPi-2a (encoded by SLC34A1 gene, mediates 70% of filtered phosphate) – NaPi-2c (encoded by SLC34A3 gene) – Pit-1/2 Different stoichiometries • NaPi-2a is electrogenic – Stoichiometry is 3:1 Na:P • NaPi-2c is eletroneutral – Stoichiometry is 2:1 • Both cotransporters show similar affinity fo Na (-50mM) and Pi (<0.1mM) • Why is stoichiometry important? – It allows the favorable inward gradient to drive continued phosphate uptake despite a falling tubular fluid phosphate concentration The role of NHERF-1 • Sodium-hydrogen exchanger regulatory factor 1 protein • Interacts with C-terminal tail of NaPi-2a and NaPi-2c • Plays an important role in the trafficking and transciptional regulation NHERF-1 Cont • Recent study by Karim et al in NEJM from September 2008 • Gene sequencing of patients with renal stones, bone demineralization, or both (usual causes such as hyperparathyroidism were excluded) • Three different NHERF1 mutations in 7 patients, which had a significantly lower renal phosphate reabsorption capacities than patients with wild type NHERF1 – Greater cAMP stimulation and greater inhibition of phosphate transport in the presence of PTH Phosphatonins • Term was coined in 1994 to describe a circulating phosphaturic factor present in patient with oncogenic or Tumor-induced osteomalacia – – – – Hypophosphatemia Renal phosphate wasting Reduced 1,25 Vit D Osteomalacia • All resolved after removal of the tumor • Include: – – – – • Fibroblast growth factor 23 (FGF-23) FGF-7 matrix extracellular phosphoglycoprotein (MEPE) secreted frizzled-related protein 4 (sFRP-4) Phosphatonins downregulate renal phosphate reabsorption at least in part by decreasing the abundance of apical sodium/phosphate co-transporters in the proximal tubule (both NaPi-2a and NaPi-2c) FGF-23 • Elevated in the following disorders (phenotypically similar to TIO): – – – – X-linked hypophosphatemic rickets (XLH) Autosomal dominant hypophosphatemic rickets (ADHR) Autosomal recessive hypophosphatemia (ARHP) Renal failure (correlates with decline in GFR as well as elevated phos) <- phenotypically different from TIO • Reduced in Tumoral calcinosis, a disorder characterized by: – Hyperphosphatemia – Reduced fractional excretion of phosphate – Ca phosphate deposits in soft tissues FGF-23 • Secreted, circulating 32kDa protein • Predominantly expressed in osteocytes in the bone and endothelial cells of bone marrow and thymus • Interacts with FGF receptors that belong to type 1 transmembrane phosphotyrosine kinase receptors (MAPK/ERK1-2) • Requires Klotho as a co-factor for receptor activation – Klotho gene encodes a single-pass membrane protein, homologous to B-glucosidase – Klotho-deficient mice have a phenotype similar to FGF-23 null mice FGF 23