Analysis of pseudoxanthoma elasticum-causing missense mutants of ABCC6 in vivo; pharmacological correction of the mislocalized proteins.
Analysis of pseudoxanthoma elasticum-causing missense mutants of ABCC6 in vivo; pharmacological correction of the mislocalized proteins.
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DOI：
10.1038/jid.2013.482
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发表时间：
2014-04
影响因子：
6.5
通讯作者：
Varadi, Andras
中科院分区：
文献类型：
--
作者：
Pomozi, Viola;Brampton, Christopher;Fueloep, Krisztina;Chen, Li-Hsieh;Apana, Ailea;Li, Qiaoli;Uitto, Jouni;Le Saux, Olivier;Varadi, Andras
ABCC6基因突变导致弹性假性黄瘤（PXE）和婴儿期全身性动脉钙化（GACI）的软组织钙化。PXE的特点是皮肤、眼部和心血管组织中弹性纤维的晚发性和进行性矿化。GACI患者表现出更严重的，通常是产前动脉钙化。我们利用Sf9细胞检测了10种常见的致病ABCC6错义突变体的转运活性，表征了MDCKII细胞和小鼠肝脏的亚细胞定位，并测试了斑马鱼的表型拯救。我们的目的是鉴定具有保留转运活性但质膜定位不当的突变体，以便通过化学伴侣4-苯基丁酸酯（4-PBA）进行救援。其中7个突变体具有运输能力，但在小鼠肝脏中定位错误。观察到的MDCKII细胞与小鼠肝脏中突变体的细胞定位差异强调了这种二维体外细胞系统的局限性。ABCC6突变体的功能在斑马鱼中进行了测试，发现对morpholino诱导的表型有最小程度的恢复。然而，被批准临床使用的药物4-PBA恢复了4个ABCC6突变体（R1114P、S1121W、Q1347H、R1314W）的质膜定位，这表明等位基因特异性治疗可能对PXE和GACI患者有用。
Mutations in the ABCC6 gene cause soft tissue calcification in pseudoxanthoma elasticum (PXE) and in some patients generalized arterial calcification of infancy (GACI). PXE is characterized by late-onset and progressive mineralization of elastic fibers in dermal, ocular and cardiovascular tissues. GACI patients present a more severe, often prenatal arterial calcification. We have tested ten frequent disease-causing ABCC6 missense mutants for the transport activity using Sf9 cells, characterized the subcellular localization in MDCKII cells and in mouse liver, and tested the phenotypic rescue in zebrafish. We aimed at identifying mutants with preserved transport activity but with improper plasma membrane localization for rescue by the chemical chaperone 4-phenylbutyrate (4-PBA). Seven of the mutants were transport-competent but mislocalized in mouse liver. The observed divergence in cellular localization of mutants in MDCKII cells vs. mouse liver underlined the limitations of this two-dimensional in vitro cell system. The functionality of ABCC6 mutants was tested in zebrafish and minimal rescue of the morpholino-induced phenotype was found. However, 4-PBA, a drug approved for clinical use, restored the plasma membrane localization of four ABCC6 mutants (R1114P, S1121W, Q1347H, R1314W), suggesting that allele-specific therapy may be useful for selected patients with PXE and GACI.
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DOI：
10.1152/ajpcell.2000.278.2.c259
发表时间：
2000-02-01
影响因子：
5.5
作者：
Rubenstein, RC;Zeitlin, PL
通讯作者：
Zeitlin, PL
影响因子：
6.5
作者：
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通讯作者：
McCormick, Sally P. A.
影响因子：
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作者：
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Kulseth, Mari A.
影响因子：
13.5
作者：
Hayashi, Hisamitsu;Sugiyama, Yuichi
通讯作者：
Sugiyama, Yuichi
影响因子：
3.6
作者：
Hayashi, Hisamitsu;Sugiyama, Yuichi
通讯作者：
Sugiyama, Yuichi