Table_1_Regulation of CIRP by genetic factors of SP1 related to cold sensitivity.docx

Cold-inducible RNA-binding-protein (CIRP) is a cold shock protein that plays a protective role in genotoxic stress response. CIRP modulates inflammation in human diseases, inhibits cell proliferation, and protects cells from genotoxic damage during cellular stress. The mild cold responsive element and specificity protein 1 (SP1) play a role in Cirp expression at low temperatures. Although previous studies have provided insights into the immune functions of SP1 or CIRP, the mechanisms by which CIRP and SP1 mediate inflammatory responses remain largely unknown. Therefore, in the current study, we examined whether Cirp expression is affected by genetic factors related to temperature sensitivity as well as under low temperature. We performed a genome-wide association study on cold sensitivity in 2,000 participants. Fifty-six genome-wide significant trait-locus pairs were identified (p<1×10-5, false discovery rate < 0.05). Among these variants, rs1117050 and rs11170510 had a strong linkage disequilibrium (r2 > 0.8) relationship and expression quantitative trait locus-associated signals with the nearest Sp1 gene. We confirmed that the minor alleles of rs11170510 and rs58123204 were associated with increased Sp1 expression. Additionally, Sp1 overexpression led to CIRP translocation from the nucleus to the cytoplasm. CIRP protein levels increased in serum samples that had minor alleles of rs11170510 and rs58123204. Levels of various pro-inflammatory cytokines were also significantly increased in human peripheral blood mononuclear cells with minor alleles of rs11170510 and rs58123204. These results suggest that genetic factors related to cold sensitivity regulate CIRP expression and function and provide valuable insights into prediction of potential diseases through analysis of inherent genetic factors in humans.

冷诱导RNA结合蛋白（Cold-inducible RNA-binding-protein, CIRP）是一类在遗传毒性应激反应中发挥保护作用的冷休克蛋白。CIRP可调控人类疾病中的炎症反应、抑制细胞增殖，并在细胞应激过程中保护细胞免受遗传毒性损伤。轻度冷应答元件（mild cold responsive element）与特异性蛋白1（specificity protein 1, SP1）在低温条件下参与Cirp基因的表达调控。尽管既往研究已对SP1或CIRP的免疫功能提供了一定见解，但CIRP与SP1介导炎症反应的具体分子机制仍未完全阐明。因此，本研究旨在探讨CIRP的表达是否受温度敏感性相关遗传因素以及低温环境的调控。我们对2000名受试者的冷敏感性开展了全基因组关联研究，共鉴定得到56个全基因组水平显著的性状-位点关联对（p<1×10^-5，错误发现率<0.05）。在上述变异位点中，rs1117050与rs11170510存在强连锁不平衡（linkage disequilibrium, r²>0.8）关系，且二者与邻近SP1基因存在表达数量性状位点（expression quantitative trait locus, eQTL）关联信号。我们证实，rs11170510与rs58123204的次要等位基因与SP1表达上调显著相关。此外，SP1过表达可导致CIRP从细胞核转位至细胞质。携带rs11170510与rs58123204次要等位基因的血清样本中，CIRP蛋白水平显著升高。在携带上述两个次要等位基因的人类外周血单个核细胞中，多种促炎细胞因子的表达水平亦显著上调。本研究结果表明，冷敏感性相关遗传因素可调控CIRP的表达与功能，为通过分析人类固有遗传因素预测潜在疾病提供了有价值的理论依据。