title: Cellular Pathophysiology of an Adrenal Adenoma-Associated Mutant of the Plasma Membrane Ca(2+)-ATPase ATP2B3.
abstract: Adrenal aldosterone-producing adenomas (APAs) are a main cause for primary aldosteronism leading to arterial hypertension. Physiologically, aldosterone production in the adrenal gland is stimulated by angiotensin II and high extracellular potassium. These stimuli lead to a depolarization of the plasma membrane and, as a consequence, an increase of intracellular Ca(2+). Mutations of the plasma membrane Ca(2+)-ATPase ATP2B3 have been found in APAs with a prevalence of 0.6%-3.1%. Here, we investigated the effects of the APA-associated ATP2B3(Leu425_Val426del) mutation in adrenocortical NCI-H295R and human embryonic kidney (HEK-293) cells. Ca(2+) measurements revealed a higher basal Ca(2+) level in cells expressing the mutant ATP2B3. This rise in intracellular Ca(2+) was even more pronounced under conditions with high extracellular Ca(2+) pointing to an increased Ca(2+) influx associated with the mutated protein. Furthermore, cells with the mutant ATP2B3 appeared to have a reduced capacity to export Ca(2+) suggesting a loss of the physiological pump function. Surprisingly, expression of the mutant ATP2B3 caused a Na(+)-dependent inward current that strongly depolarized the plasma membrane and compromised the cytosolic cation composition. In parallel to these findings, mRNA expression of the cytochrome P450, family 11, subfamily B, polypeptide 2 (aldosterone synthase) was substantially increased and aldosterone production was enhanced in cells overexpressing mutant ATP2B3. In summary, the APA-associated ATP2B3(Leu425_Val426del) mutant promotes aldosterone production by at least 2 different mechanisms: 1) a reduced Ca(2+) export due to the loss of the physiological pump function; and 2) an increased Ca(2+) influx due to opening of depolarization-activated Ca(2+) channels as well as a possible Ca(2+) leak through the mutated pump.
authors.affiliation: Medical Cell Biology (P.T., B.A., C.C., J.S., R.W., S.B.), University of Regensburg, 93053 Regensburg, Germany; and Medizinische Klinik und Poliklinik IV (Y.R., F.B.), Ludwig-Maximilians-Universität, 80336 Munich, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Tauber
authors.affiliation: Medical Cell Biology (P.T., B.A., C.C., J.S., R.W., S.B.), University of Regensburg, 93053 Regensburg, Germany; and Medizinische Klinik und Poliklinik IV (Y.R., F.B.), Ludwig-Maximilians-Universität, 80336 Munich, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Aichinger
authors.affiliation: Medical Cell Biology (P.T., B.A., C.C., J.S., R.W., S.B.), University of Regensburg, 93053 Regensburg, Germany; and Medizinische Klinik und Poliklinik IV (Y.R., F.B.), Ludwig-Maximilians-Universität, 80336 Munich, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Christ
authors.affiliation: Medical Cell Biology (P.T., B.A., C.C., J.S., R.W., S.B.), University of Regensburg, 93053 Regensburg, Germany; and Medizinische Klinik und Poliklinik IV (Y.R., F.B.), Ludwig-Maximilians-Universität, 80336 Munich, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Stindl
authors.affiliation: Medical Cell Biology (P.T., B.A., C.C., J.S., R.W., S.B.), University of Regensburg, 93053 Regensburg, Germany; and Medizinische Klinik und Poliklinik IV (Y.R., F.B.), Ludwig-Maximilians-Universität, 80336 Munich, Germany.
authors.forename: Y
authors.initials: Y
authors.lastname: Rhayem
authors.affiliation: Medical Cell Biology (P.T., B.A., C.C., J.S., R.W., S.B.), University of Regensburg, 93053 Regensburg, Germany; and Medizinische Klinik und Poliklinik IV (Y.R., F.B.), Ludwig-Maximilians-Universität, 80336 Munich, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Beuschlein
authors.affiliation: Medical Cell Biology (P.T., B.A., C.C., J.S., R.W., S.B.), University of Regensburg, 93053 Regensburg, Germany; and Medizinische Klinik und Poliklinik IV (Y.R., F.B.), Ludwig-Maximilians-Universität, 80336 Munich, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Warth
authors.affiliation: Medical Cell Biology (P.T., B.A., C.C., J.S., R.W., S.B.), University of Regensburg, 93053 Regensburg, Germany; and Medizinische Klinik und Poliklinik IV (Y.R., F.B.), Ludwig-Maximilians-Universität, 80336 Munich, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Bandulik
mesh_terms: D000311:Adrenal Glands / Q000378:metabolism; D018246:Adrenocortical Adenoma / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology*; D000450:Aldosterone / Q000378:metabolism; D000804:Angiotensin II / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D019405:Cytochrome P-450 CYP11B2 / Q000235:genetics / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000235:genetics / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism
chemical_list: D000804:Angiotensin II; D000450:Aldosterone; D012964:Sodium; D019405:Cytochrome P-450 CYP11B2; D053499:Plasma Membrane Calcium-Transporting ATPases; C506962:ATP2B3 protein, human; D002118:Calcium