A paradoxical feature of several disorders of the skeleton involves deficits in bone mineral density yet progressive increases in vascular and/or soft tissue calcifications. ENPP1 loss of function (LOF) mutations represent this combination of defects, in that both life threatening vascular calcifications in Generalized Arterial Calcification of Infancy (GACI) and a severe rickets in Autosomal Recessive Hypophosphatemic Rickets Type-2 (ARHR2) are observed with this mutation. Disease in ARHR2 is severe and symptomatic, leading to repeated fractures of the long bones, rachitic deformities, and impaired growth and development. ENPP1 LOF mutations in mice induces paradoxical mineralization characterized by increased cardiovascular and periarticular calcifications and an osteopenic bone phenotype, reflecting important aspects of both GACI and ARHR2. Here, we report that the osteopenic phenotype in Enpp1asj/asj mice results from an osteomalacia, the histologic hallmark of rickets. The osteomalacia present in Enpp1asj/asj mice occurs in the presence of normal plasma [Pi] in mice fed a high phosphate diet. Treatment of Enpp1asj/asj mice with ENPP1 enzyme replacement therapy (ERT) eliminates the osteomalacia, increases bone mineral density, and markedly improves bone strength and ductility. Our work demonstrates that ENPP1 ERT addresses the pathologic etiology of ARHR2, but phosphate supplementation does not. Combined with our previous studies demonstrating the efficacy of ENPP1-Fc suppressing lethal vascular and connective tissue calcifications, this study establishes a role for ENPP1 in paradoxical mineralization disorders and suggests a therapeutic strategy for treating these diseases. Support or Funding Information The studies were financially supported by research grants from Inozyme Pharmaceuticals Inc., and The Connecticut Bioscience Innovation Fund (to DTB). Funding to the George M O'Brien Kidney Center at Yale, NIH grant P30DK079310, supported the evaluation of plasma analytes. Histology of the proximal tibias of 5-week old mice on the high phosphate diet are displayed from each experimental cohort to compare histologic changes. Deficiency in ENPP1 resulted in morphologically apparent reductions in trabecular bone and markedly thinner growth plates in the Enpp1asj/asj mice. Treating Enpp1asj/asj mice with mENPP1-Fc markedly increased trabecular bone volume and growth plate thickness. WT and Enpp1asj/asj mice treated with vehicle or ENPP1-Fc were loaded to failure in three-point bending. a. Stiffness â slope of the load vs. displacement curve; Max Load â highest load reached during the test.; Total Work â The energy needed to fracture the bone. b. Post-yield deflection of experimental cohorts â defined as the amount of deformation after the yield point. *p < 0.05, **p< 0.01, ***p<0.001, ANOVA. Histomorphometry, 5 week females, High Phosphate Diet, osteoid. Genotypes/Treatment WT/PBS N=6 asj-asj/PBS n=7 asj-asj/mENPP1 n=6 BV/TV 19.23 ± 3.42 24.61 ± 6.45 20.63 ± 4.58 OV/TV 1.38 ± 0.31 2.35 ± 0.57** 1.78 ± 0.50 OV/BV 6.99 ± 1.76 9.72 ± 1.57* 8.60 ± 1.45 OS/BS 27.89 ± 4.66 42.83 ± 4.52*** 33.68 ± 6.41 OTh 3.01 ± 0.29 3.53 ± 0.29** 3.30 ± 0.12 This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
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