A naturally occurring mutation in MRP1 results in a selective decrease in organic anion transport and in increased doxorubicin resistance.

The human 190 kDa multidrug resistance protein, MRP1, is a polytopic membrane glycoprotein that confers resistance to a wide range of chemotherapeutic agents. It also transports structurally diverse conjugated organic anions, as well as certain unconjugated and conjugated compounds, in a reduced glutathione-stimulated manner. In this study, we characterized a low-frequency (<1%) naturally occurring mutation in MRP1 expected to cause the substitution of a conserved arginine with serine at position 433 in a predicted cytoplasmic loop of the protein. Transport experiments with membrane vesicles prepared from transfected human embryonic kidney cells and HeLa cells revealed a two-fold reduction in the ATP-dependent transport of the MRP1 substrates, leukotriene C4 (LTC4) and oestrone sulphate. Kinetic analysis showed that this reduction was due to a decrease in Vmax for both substrates but Km was unchanged. In contrast, 17beta-oestradiol-17beta-(D-glucuronide) transport by the Arg433Ser mutant MRP1 was similar to that by wild-type MRP1. Fluorescence confocal microscopy showed that the mutant MRP1 was routed correctly to the plasma membrane. In contrast to the reduced LTC4 and oestrone sulphate transport, stably transfected HeLa cells expressing Arg433Ser mutant MRP1 were 2.1-fold more resistant to doxorubicin than cells expressing wild-type MRP1, while resistance to VP-16 and vincristine was unchanged. These results provide the first example of a naturally occurring mutation predicted to result in an amino acid substitution in a cytoplasmic region of MRP1 that shows an altered phenotype with respect to both conjugated organic anion transport and drug resistance.

The thrust plate prosthesis in patients with aseptic osteonecrosis of the femoral head.

BACKGROUND: The thrust plate prosthesis (TPP) is an implant with a metaphyseal fixation at the proximal femur that transmits the load forces of the hip onto the femoral neck. The osseous incorporation of the TPP and the adaptation of the bone to this force transmission depend on the bone quality, which is reduced to minor vitality and stability in patients with osteonecrosis of the femoral head. Depending on the etiology of the femoral head necrosis, the TPP might lead to early failures. METHODS: In a prospective study, 63 patients with 72 cementless TPP due to femoral head osteonecrosis were examined. A clinical and radiological evaluation was performed preoperatively, 3 and 6 months postoperatively, and every year thereafter. The average follow-up period was 4.8+/-1.3 years with a minimum of 3 years. The pathogenesis of femoral head necrosis included alcoholism (n=19), subsequent to renal transplantation (n=11), during cortisone therapy of other dyscrasia (n=9), preceding a polychemotherapy (n=4), diabetes (n=3), sickle cell anemia (n=1), and idiopathic osteonecrosis (n=25). RESULTS: The Harris Hip Score increased continuously from 50.0 points beyond 79.8 points after 3 months to 86.8 points within the 1st year, and subsequently remained stable at this level. Revision was necessary in six cases (8.3%). Of these, three had an aseptic loosening of the implant: 2 cases with renal transplantation and 1 of alcoholism with an extension of the necrotic area to the seating of the TPP. The other three patients showed septic implant loosenings: 2 cases with renal transplantation and 1 of alcoholism. Radiolucent lines were found in 9 cases (12.5%), mostly in zones 1 and 2 underneath the TPP. Of these, 1 with an idiopathic osteonecrosis was assessed to be radiologically loosened. The overall failure rate was 9.7%, with a proportion of 36.4% in patients with renal transplantation. Excluding this specific patient group, the failure rate was 4.9%. CONCLUSIONS: Femoral head necrosis following renal transplantation and extension of the necrotic area into the femoral neck are contraindications for TPP. Excluding these patients, the TPP shows comparable mid-term results to cementless stemmed prostheses and supplies advantages especially for younger patients, because of its metaphyseal, bone-preserving fixation. However, evaluation of the clinical impact of the TPP in comparison with other cementless femoral stem systems requires long-term examinations in the future.