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

NCI方案：10247“一项pevonedistat联合阿扎胞苷与阿扎胞苷治疗成人复发性或难治性急性髓性白血病的随机化II期试验”; 10249“Ixlidine和Pevonedistat治疗复发性/难治性多发性骨髓瘤患者：一项Ib期试验”; 10246“Pevonedistat和贝利司他治疗复发性/难治性急性髓性白血病或骨髓增生异常综合征”; 10266“Pevonedistat联合卡铂和紫杉醇治疗既往接受过免疫治疗的晚期NSCLC的II期研究”; ADVL 1712-“Pevonedistat联合阿扎胞苷、氟达拉滨和阿糖胞苷治疗复发性或难治性急性髓性白血病儿童、青少年和年轻成人的可行性试验” NAE激活泛素样蛋白NEDD 8与CRL结合，因此调节CRL底物蛋白的蛋白酶体破坏。 NAE抑制防止CRL底物的降解（例如，NRF 2，CDT 1），导致其积累。 为了开发药效学试验，步行者et al.（2011）开发了一种基于RT-PCR的试验，该试验使用了多名健康志愿者的血液样本，这些志愿者接受了一系列MLN 4924（pevonedistat）浓度的离体处理。 八个基因（包括NRF 2调节的基因NQO 1和SLC 7A 11）在全血中显示出稳健的诱导（>3倍）。 本试验旨在评估MLN 4924靶点结合（通过RT-PCR测定全血中NRF 2调节的NQO 1和SLC 7A 11基因表达增加）是否为MLN 4924活性的生物标志物。

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.