Activation of Calcium-Sensing Receptor increases intracellular calcium and decreases cAMP and mTOR in PKD1 deficient cells

被引：33

作者：

Di Mise, Annarita ^{[1
]}

Tamma, Grazia ^{[1
,5
]}

Ranieri, Marianna ^{[1
]}

Centrone, Mariangela ^{[1
]}

van den Heuvel, Lambertus ^{[2
]}

Mekahli, Djalila ^{[3
,4
]}

Levtchenko, Elena N. ^{[3
,4
]}

Valenti, Giovanna ^{[1
,5
,6
]}

机构：

[1] Univ Bari, Dept Biosci Biotechnol & Biopharmaceut, I-70125 Bari, Italy

[2] Radboud Univ Nijmegen, Med Ctr, Dept Pediat Nephrol, NL-6525 HP Nijmegen, Netherlands

[3] Univ Hosp Gasthuisberg, Dept Pediat Nephrol, B-3000 Leuven, Belgium

[4] Univ Leuven, KU Leuven, Dept Dev & Regenerat, B-3000 Leuven, Belgium

[5] Ist Nazl Biostrutture & Biosistemi, I-00136 Rome, Italy

[6] Univ Bari, Ctr Excellence Comparat Genom CEGBA, I-70125 Bari, Italy

来源：

SCIENTIFIC REPORTS | 2018年 / 8卷

关键词：

POLYCYSTIC KIDNEY-DISEASE; EPITHELIAL-CELLS; LIVER-DISEASE; CA2+ RELEASE; MODEL; VASOPRESSIN; RAPAMYCIN; PATHWAY; GROWTH; PHOSPHORYLATION;

D O I：

10.1038/s41598-018-23732-5

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Clinical and fundamental research suggest that altered calcium and cAMP signaling might be the most proximal events in ADPKD pathogenesis. Cells from ADPKD cysts have a reduced resting cytosolic calcium [Ca2+](i) and increased cAMP levels. CaSR plays an essential role in regulating calcium homeostasis. Its activation is associated with [Ca2+](i) increase and cAMP decrease, making CaSR a possible therapeutic target. Human conditionally immortalized Proximal Tubular Epithelial cells (ciPTEC) with stable knockdown of PKD1 (ciPTEC-PC1KD) and ciPTEC generated from an ADPKD1 patient (ciPTEC-PC1Pt) were used as experimental tools. CaSR functional expression was confirmed by studies showing that the calcimimetic NPS-R568 induced a significant increase in [Ca2+](i) in ciPTEC-PC1KD and ciPTEC-PC1Pt. Resting [Ca2+](i) were significantly lower in ciPTEC-PC1KD with respect to ciPTECwt, confirming calcium dysregulation. As in native cyst cells, significantly higher cAMP levels and mTOR activity were found in ciPTEC-PC1KD compared to ciPTECwt. Of note, NPS-R568 treatment significantly reduced intracellular cAMP and mTOR activity in ciPTEC-PC1KD and ciPTEC-PC1Pt. To conclude, we demonstrated that selective CaSR activation in human ciPTEC carrying PKD1 mutation increases [Ca2+](i), reduces intracellular cAMP and mTOR activity, reversing the principal dysregulations considered the most proximal events in ADPKD pathogenesis, making CaSR a possible candidate as therapeutic target.

引用

页数：14

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