Department of Pediatrics, University of California San Francisco, Oakland, CA, USA.
Howard Hughes Medical Institute, Department of Microbiology and Immunology, University of California San Francisco, San Francisco, CA, USA.
Genome Med. 2024 Jul 26;16(1):93. doi: 10.1186/s13073-024-01366-9.
Statins lower circulating low-density lipoprotein cholesterol (LDLC) levels and reduce cardiovascular disease risk. Though highly efficacious in general, there is considerable inter-individual variation in statin efficacy that remains largely unexplained.
To identify novel genes that may modulate statin-induced LDLC lowering, we used RNA-sequencing data from 426 control- and 2 µM simvastatin-treated lymphoblastoid cell lines (LCLs) derived from European and African American ancestry participants of the Cholesterol and Pharmacogenetics (CAP) 40 mg/day 6-week simvastatin clinical trial (ClinicalTrials.gov Identifier: NCT00451828). We correlated statin-induced changes in LCL gene expression with plasma LDLC statin response in the corresponding CAP participants. For the most correlated gene identified (ZNF335), we followed up in vivo by comparing plasma cholesterol levels, lipoprotein profiles, and lipid statin response between wild-type mice and carriers of a hypomorphic (partial loss of function) missense mutation in Zfp335 (the mouse homolog of ZNF335).
The statin-induced expression changes of 147 human LCL genes were significantly correlated to the plasma LDLC statin responses of the corresponding CAP participants in vivo (FDR = 5%). The two genes with the strongest correlations were zinc finger protein 335 (ZNF335 aka NIF-1, rho = 0.237, FDR-adj p = 0.0085) and CCR4-NOT transcription complex subunit 3 (CNOT3, rho = 0.233, FDR-adj p = 0.0085). Chow-fed mice carrying a hypomorphic missense (R1092W; aka bloto) mutation in Zfp335 had significantly lower non-HDL cholesterol levels than wild-type C57BL/6J mice in a sex combined model (p = 0.04). Furthermore, male (but not female) mice carrying the Zfp335 allele had significantly lower total and HDL cholesterol levels than wild-type mice. In a separate experiment, wild-type mice fed a control diet for 4 weeks and a matched simvastatin diet for an additional 4 weeks had significant statin-induced reductions in non-HDLC (-43 ± 18% and -23 ± 19% for males and females, respectively). Wild-type male (but not female) mice experienced significant reductions in plasma LDL particle concentrations, while male mice carrying Zfp335 allele(s) exhibited a significantly blunted LDL statin response.
Our in vitro and in vivo studies identified ZNF335 as a novel modulator of plasma cholesterol levels and statin response, suggesting that variation in ZNF335 activity could contribute to inter-individual differences in statin clinical efficacy.
他汀类药物可降低循环中的低密度脂蛋白胆固醇（LDLC）水平并降低心血管疾病风险。尽管他汀类药物总体上非常有效，但个体之间的他汀类药物疗效存在很大差异，这在很大程度上仍未得到解释。
为了确定可能调节他汀类药物诱导的 LDLC 降低的新基因，我们使用了来自欧洲和非裔美国人血统的胆固醇和药物遗传学（CAP）40mg/天 6 周辛伐他汀临床试验（ClinicalTrials.gov 标识符：NCT00451828）中 426 个对照和 2µM 辛伐他汀处理的淋巴母细胞系（LCL）的 RNA 测序数据。我们将他汀类药物诱导的 LCL 基因表达变化与相应 CAP 参与者的血浆 LDLC 他汀类药物反应相关联。对于鉴定出的最相关基因（ZNF335），我们通过比较野生型小鼠和携带 Zfp335 （ZNF335 的小鼠同源物）部分功能丧失（部分丧失功能）错义突变的小鼠的血浆胆固醇水平、脂蛋白谱和脂质他汀类药物反应，在体内进行了随访。
147 个人类 LCL 基因的他汀类药物诱导表达变化与相应 CAP 参与者的体内血浆 LDLC 他汀类药物反应显著相关（FDR=5%）。相关性最强的两个基因是锌指蛋白 335（ZNF335 又名 NIF-1，rho=0.237，FDR 调整 p=0.0085）和 CCR4-NOT 转录复合物亚基 3（CNOT3，rho=0.233，FDR 调整 p=0.0085）。在 Chow 喂养的小鼠中，携带 Zfp335 的功能丧失（R1092W；又名 bloto）错义突变的小鼠的非 HDL 胆固醇水平明显低于野生型 C57BL/6J 小鼠（p=0.04）。此外，携带 Zfp335 等位基因的雄性（但不是雌性）小鼠的总胆固醇和 HDL 胆固醇水平明显低于野生型小鼠。在另一个实验中，野生型小鼠接受对照饮食 4 周，然后接受匹配的辛伐他汀饮食 4 周，非 HDL-C 有显著的他汀类药物诱导降低（雄性分别为-43±18%和-23±19%，雌性）。野生型雄性（但不是雌性）小鼠的血浆 LDL 颗粒浓度显著降低，而携带 Zfp335 等位基因的雄性小鼠的 LDL 他汀类药物反应明显减弱。
我们的体外和体内研究将 ZNF335 鉴定为一种新的血浆胆固醇水平和他汀类药物反应调节剂，表明 ZNF335 活性的差异可能导致他汀类药物临床疗效的个体间差异。
