As3MT基因多态性对As2O3在治疗急性早幼粒细胞白血病中的代谢及疗效影响研究

Arsenic has been used successfully in clinical trials for treating acute promyelocytic leukemia (APL), but the potential side effects of arsenic treatment limit its application. It has important significance to study the metabolism of As2O3 in vivo. At present, many studies focus on effects of trivalent arsenic methyltransferase (As3MT) on arsenic metabolism. Little attention has been paid to the relationship between genetic polymorphism of As3MT and APL patients treated with As2O3. In our previous study, we analyzed the genetic polymorphism of As3MT in 100 APL patients, and found that two single nucleotide polymorphisms (SNPs), As3MT (rs3740390 and rs10748835) were related to the clinical remission of APL and metabolism of As2O3. Therefore, we hypothesized that the As3MT genotype determines the treatment efficacy and side effects of As2O3. To study the above hypothesis, we elucidate the effect and mechanism of genetic polymorphisms of As3MT in the treatment of APL with As2O3 in vitro and in vivo, which are evaluated using the technology of pharmaceutical analysis, bioinformatics, pathology, cytology, and molecular biology. In conclusion, the study will have new insight into individual treatment of APL patients with As2O3, which can maximize the treatment efficacy and minimize side effects.

As2O3治疗急性早幼粒细胞白血病的毒副作用限制了其应用，研究As2O3的体内代谢具有重要意义。目前，三价砷甲基转移酶（As3MT）与砷代谢成为研究的热点，但其基因多态性与As2O3治疗APL患者后代谢及疗效的相关性却未见报道。我们在前期工作中：对100例APL患者As3MT基因多态性进行了比较分析，发现两个位点（rs3740390、rs10748835）的基因型与As2O3的代谢及疗效具有相关性。因此，我们提出假设：As3MT不同位点的基因型决定了As2O3代谢产物差异，从而产生相应的疗效及并发症。为了验证我们的假设拟采用临床患者标本、动物及细胞模型，运用药物分析学、生物信息学、病理学、细胞学及分子生物学等技术方法，探寻As3MT参与As2O3治疗APL的关键位点及基因型，明确其调控的作用机制。本课题的完成有助于APL患者个体化As2O3治疗，对提高治愈率、预防并发症具有重要的临床价值。

亚砷酸注射液（As2O3）是我国自主研发对急性早幼粒细胞白血病（APL）疗效显著的国家二类新药。As2O3可生成三价无机砷（AsIII）、一甲基砷（MMAⅢ、MMAⅤ）、二甲基砷（DMAⅢ、DMAⅤ）和五价无机砷（AsⅤ）不同形态砷代谢产物，其毒性差异显著。As2O3疗效及毒副作用与砷化物种类及浓度息息相关，砷代谢研究意义重大。砷甲基转移酶(As3MT）是砷代谢的重要酶，其存在基因多态性。.建立了As2O3治疗的APL患者血浆、红细胞、尿液、粒细胞、脑脊液中砷及其代谢产物的HPLC-HG-AFS及HPLC-ICP-MS检测方法，明确了各形态砷化物在APL患者体内浓度特征、分布规律及药代动力学特点等，探讨了各种临床因素对砷代谢的影响，发现了APL患者体内砷及其代谢产物浓度对As2O3疗效的预测作用。.证明了APL患者的As3MT基因多态性可导致砷的个体甲基化代谢能力产生差异，造成体内砷及其代谢产物浓度、甲基化物水平的差异，进而影响As2O3治疗APL的疗效和毒副作用的发生。As3MT 14215(rs3740390) CT+TT、35991(rs10748835) AA+35587(rs11191453) TC+CC基因型携带者具有更高的DMA%，而14215(rs3740390)CC基因型携带者DMA%和二次甲基化指数（SMI）明显偏低，ALT和AST水平明显升高，肝损伤发生率高；As3MT 14215(rs3740390) CC基因型携带者甲基化代谢能力差，且高白细胞血症发生率高。通过As3MT SNP位点检测，可提前预测毒副反应的发生。结合体内砷化物测定与基因多态性检测，改善As2O3疗效，降低毒副作用，实现APL精准治疗。.探究了As2O3治疗的APL患者体内砷化物水平及分布与毒副作用的相关性，评估大鼠口服As2O3后血液和肝、肾等组织中砷及其代谢产物的累积与毒性，为后续阐明毒副作用机制研究奠定基础。