NQO1/SLC711A Assay, Total RNA isolation and running custom TaqMan cards (TLDA) assay on up to 486 PAXgene Blood RNA Samples

美国国立卫生研究院方案(S)：10247“在成人复发性或难治性急性髓细胞白血病中，培氟地沙联合阿扎替丁与阿扎替丁治疗的随机II期试验”；10249“伊沙唑米布和培伐地沙治疗复发/难治性多发性骨髓瘤患者：Ib期试验”；10246“培氟地沙和倍立得在复发/难治性急性髓系白血病或骨髓增生异常综合征中的应用”；10266“培伐地沙联合卡铂和紫杉醇治疗晚期非小细胞肺癌的2期研究”；ADVL1712--“在儿童、青少年和青年复发性或难治性骨髓白血病患者中给予Pevonedexat联合卡铂、氟达拉滨和阿糖胞苷的可行性试验”；ADVL1712--“Pevonedexat与卡铂和紫杉醇联合应用于儿童、青少年和青年复发或难治性骨髓白血病的可行性试验”。 Nae激活泛素样蛋白NEDD8与CRL结合，从而调节CRL底物蛋白的蛋白酶体破坏。抑制NAE可防止CRL底物(如NRF2、CDT1)的降解，导致它们的积累。为了开发一种药效学分析，Walker等人。(2011)开发了一种基于RT-PCR的检测方法，使用了多名健康志愿者的血液样本，这些志愿者在体外接受了一系列MLN4924(PevoneDistat)浓度的治疗。8个基因(包括NRF2调节基因NQO1和SLC7A11)在全血中显示出强大的诱导性(&gt；3倍)。本试验的目的是评估MLN4924的靶标接入量是否是MLN4924活性的生物标志物。通过RT-PCR检测，MLN4924靶向结合是由NRF2调节的全血中NQO1和SLC7A11基因表达的增加。

NCI Protocol(s): 10247 “A randomized phase II trial of pevonedistat with Azacitidine versus Azacitidine in adult relapsed or refractory acute myeloid leukemia”; 10249 “Ixazomib and Pevonedistat in Relapsed/Refractory Multiple Myeloma Patients: A Phase Ib Trial”; 10246 “Pevonedistat and belinostat in relapsed/refractory acute myeloid leukemia or myelodysplastic syndrome”; 10266 “A Phase 2 Study of Pevonedistat in Combination with Carboplatin and Paclitaxel in Advanced NSCLC Previously Treated with Immunotherapy”; ADVL1712 – “A Feasibility Trial of Pevonedistat Given in Combination with Azacitidine, Fludarabine, and Cytarabine, in Children, Adolescents, and Young Adults with Relapsed or Refractory Acute Myeloid Leukemia” NAE activates the ubiquitin-like protein NEDD8 for conjugation to CRLs and, therefore, regulates the proteasomal destruction of CRL substrate proteins. NAE inhibition prevents degradation of CRL substrates (e.g., NRF2, CDT1), leading to their accumulation. To develop a pharmacodynamic assay, Walker et al. (2011) developed a RT-PCR-based assay using blood samples from multiple healthy volunteers that were treated ex vivo with a range of MLN4924 (pevonedistat) concentrations. Eight genes (including the NRF2-regulated genes NQO1 and SLC7A11) displayed a robust induction (>3-fold) in whole blood. This assay is aimed at assessing whether MLN4924 target engagement, as measured by increased NRF2-regulated NQO1 and SLC7A11 gene expression in whole blood by RT-PCR, is a biomarker of MLN4924 activity.