Department of Internal Medicine, Hallym University Medical Center, Chuncheon Sacred Heart Hospital, Chuncheon 24253, Republic of Korea.
Center for Medical Innovation, Biomedical Research Institute, Seoul National University Hospital, Seoul 03080, Republic of Korea.
Biomolecules. 2024 Jul 7;14(7):806. doi: 10.3390/biom14070806.
Autosomal dominant polycystic kidney disease (ADPKD) is the most prevalent genetic kidney disorder. While metformin has demonstrated the ability to inhibit cyst growth in animal models of ADPKD via activation of adenosine monophosphate-activated protein kinase (AMPK), its effectiveness in humans is limited due to its low potency. This study explored the impact of HL156A, a new and more potent AMPK activator, in a mouse model of ADPKD.
To investigate whether HL156A inhibits the proliferation of renal cyst cells in ADPKD in vitro, exogenous human telomerase reverse transcriptase (hTERT)-immortalized renal cyst cells from ADPKD patients were treated with HL156A, and an MTT (dimethylthiazol-diphenyltetrazolium bromide) assay was performed. To assess the cyst-inhibitory effect of HL156A in vivo, we generated conditional knockout (KO) mice with aquaporin 2 (AQP2)-Cre, which selectively expresses Cre recombinase in the collecting duct. The effectiveness of HL156A in inhibiting cyst growth and improving renal function was confirmed by measuring the number of cysts and blood urea nitrogen (BUN) levels in the collecting duct-specific KO mice.
When cyst cells were treated with up to 20 µM of metformin or HL156A, HL156A reduced cell viability by 25% starting at a concentration of 5 µM, whereas metformin showed no effect. When AQP2-Cre male mice were crossed with female mice, and when AQP2-Cre female mice were crossed with male mice, the number of litters produced by both groups was comparable. In collecting duct-specific KO mice, HL156A was found to inhibit cyst growth, reducing both the number and size of cysts. Furthermore, it was confirmed that kidney function improved as HL156A treatment led to a reduction in elevated BUN levels. Lastly, it was observed that the increase in AMPK phosphorylation induced by HL156A decreased ERK phosphorylation and α-SMA expression.
HL156A has potential as a drug that can restore kidney function in ADPKD patients by inhibiting cyst growth.
常染色体显性多囊肾病（ADPKD）是最常见的遗传性肾脏疾病。二甲双胍通过激活腺苷单磷酸激活蛋白激酶（AMPK）已被证明能够抑制 ADPKD 动物模型中的囊肿生长，但由于其效力较低，其在人类中的效果有限。本研究探讨了新型、更有效的 AMPK 激活剂 HL156A 在 ADPKD 小鼠模型中的作用。
为了研究 HL156A 是否能抑制 ADPKD 体外肾囊肿细胞的增殖，用 HL156A 处理来自 ADPKD 患者的外源性人端粒酶逆转录酶（hTERT）永生化肾囊肿细胞，并进行 MTT（二甲基噻唑二苯基四唑溴盐）检测。为了评估 HL156A 在体内抑制囊肿的作用，我们生成了水通道蛋白 2（AQP2）-Cre 条件性敲除（KO）小鼠，该小鼠在集合管中选择性表达 Cre 重组酶。通过测量集合管特异性 KO 小鼠中的囊肿数量和血尿素氮（BUN）水平，证实了 HL156A 抑制囊肿生长和改善肾功能的有效性。
当用高达 20 μM 的二甲双胍或 HL156A 处理囊肿细胞时，HL156A 在 5 μM 浓度时开始使细胞活力降低 25%，而二甲双胍则没有效果。当 AQP2-Cre 雄性小鼠与雌性小鼠交配，或 AQP2-Cre 雌性小鼠与雄性小鼠交配时，两组的产仔数相当。在集合管特异性 KO 小鼠中，发现 HL156A 抑制囊肿生长，减少囊肿的数量和大小。此外，证实 HL156A 治疗可降低升高的 BUN 水平，从而改善肾功能。最后，观察到 HL156A 诱导的 AMPK 磷酸化增加可降低 ERK 磷酸化和 α-SMA 表达。
HL156A 具有通过抑制囊肿生长恢复 ADPKD 患者肾功能的潜力，有可能成为一种药物。
