Chloroquine increases osteoclast activity in vitro but does not improve the osteopetrotic bone phenotype of ADO2 mice.
Chloroquine increases osteoclast activity in vitro but does not improve the osteopetrotic bone phenotype of ADO2 mice.
复制标题
DOI：
10.1016/j.bone.2021.116160
复制
发表时间：
2021-12
期刊：
影响因子：
4.1
通讯作者：
Econs MJ
中科院分区：
文献类型：
--
作者：
Alam I;Gerard-O'Riley RL;Acton D;Hardman SL;Hong JM;Bruzzaniti A;Econs MJ
常染色体显性骨石症 II 型 (ADO2) 是一种破骨细胞骨吸收受损的骨病，通常由氯离子通道 7 (CLCN7) 基因的杂合错义突变引起。我们通过在 Clcn7 基因中引入敲入 (p.G213R) 突变来创建 ADO2 小鼠模型，该突变类似于人类中发现的一种常见突变 (G215R)。该突变导致纯合子小鼠出现严重的骨硬化症和致死性，但在不同遗传背景的杂合子小鼠中产生显着的表型变异，这些遗传背景与人类 ADO2 疾病的表型相似。 ADO2 是一种破骨细胞固有的疾病，溶酶体酶和蛋白质对于破骨细胞活性至关重要。已知氯喹 (CQ) 会影响溶酶体运输、细胞内信号传导以及溶酶体和囊泡 pH 值，表明它可能会改善 ADO2 破骨细胞功能。我们使用来自野生型（WT或ADO2+/+）和ADO2杂合子（ADO2+/-）小鼠的破骨细胞在细胞培养研究中测试了这一假设，发现CQ及其代谢物去乙基氯喹（DCQ）在体外显着增加ADO2+/-破骨细胞的骨吸收活性，而ADO2+/+破骨细胞的骨吸收仅通过DCQ增加。此外，我们在 129 背景下利用我们独特的 ADO2 动物模型来确定 CQ 对 ADO2 治疗的效果。 8 周龄雌性 ADO2 小鼠通过饮用水接受 5 剂 CQ（1、2.5、5、7.5 和 10 mg/kg BW/天）治疗 6 个月。通过纵向体内 DXA 和显微 CT 在基线、3 个月和 6 个月时分析骨矿物质密度和骨微结构。还在这些时间点分析了血清骨生物标志物（CTX、TRAP 和 P1NP）。与对照组（仅水）相比，ADO2 小鼠中测试剂量的 CQ 治疗未能产生 aBMD、BMC（全身、股骨和脊柱）和小梁 BV/TV（远端股骨）的任何显着变化。此外，这些小鼠的骨生物标志物水平并未因 CQ 治疗而发生显着变化。我们的研究结果表明，虽然 CQ 增加了体外破骨细胞活性，但它并没有改善 ADO2 杂合子小鼠的骨硬化骨表型。
Autosomal Dominant Osteopetrosis type II (ADO2) is a bone disease of impaired osteoclastic bone resorption that usually results from heterozygous missense mutations in the chloride channel 7 (CLCN7) gene. We created mouse models of ADO2 by introducing a knock-in (p.G213R) mutation in the Clcn7 gene, which is analogous to one of the common mutations (G215R) found in humans. The mutation leads to severe osteopetrosis and lethality in homozygous mice but produces substantial phenotypic variability in heterozygous mice on different genetic backgrounds that phenocopy the human disease of ADO2. ADO2 is an osteoclast-intrinsic disease, and lysosomal enzymes and proteins are critical for osteoclast activity. Chloroquine (CQ) is known to affect lysosomal trafficking, intracellular signaling and the lysosomal and vesicular pH, suggesting it might improve ADO2 osteoclast function. We tested this hypothesis in cell culture studies using osteoclasts derived from wild-type (WT or ADO2+/+) and ADO2 heterozygous (ADO2+/−) mice and found that CQ and its metabolite desethylchloroquine (DCQ), significantly increased ADO2+/− osteoclasts bone resorption activity in vitro, whereas bone resorption of ADO2+/+ osteoclasts was increased only by DCQ. In addition, we exploited our unique animal model of ADO2 on 129 background to identify the effect of CQ for the treatment of ADO2. Female ADO2 mice at 8 weeks of age were treated with 5 doses of CQ (1, 2.5, 5, 7.5 and 10 mg/kg BW/day) via drinking water for 6 months. Bone mineral density and bone micro-architecture were analyzed by longitudinal in-vivo DXA and micro-CT at baseline, 3 and 6 months. Serum bone biomarkers (CTX, TRAP and P1NP) were also analyzed at these time points. CQ treatment at the doses tested failed to produce any significant changes of aBMD, BMC (whole body, femur and spine) and trabecular BV/TV (distal femur) in ADO2 mice compared to the control group (water only). Further, levels of bone biomarkers were not significantly changed due to CQ treatment in these mice. Our findings indicate that while CQ increased osteoclast activity in vitro, it did not improve the osteopetrotic bone phenotypes in ADO2 heterozygous mice.
登录
查看更多内容
影响因子：
5.5
作者：
Delaisse JM;Søe K;Andersen TL;Rojek AM;Marcussen N
通讯作者：
Marcussen N
影响因子：
5.1
作者：
He, Shasha;Zhou, Qian;Yan, Fuhua
通讯作者：
Yan, Fuhua
影响因子：
1.6
作者：
JOHNSTON, CC;LAVY, N;DEISS, WP
通讯作者：
DEISS, WP
影响因子：
11.4
作者：
Leisle, Lilia;Ludwig, Carmen F.;Stauber, Tobias
通讯作者：
Stauber, Tobias
DOI：
10.4269/ajtmh.1987.36.213
发表时间：
1987-03-01
影响因子：
3.3
作者：
KROGSTAD, DJ;SCHLESINGER, PH
通讯作者：
SCHLESINGER, PH