Quantifying PBMCs and Nrf2 Transcripts

PBMCs were isolated from blood collected at each timepoint. Day 28 collections were missing for the 15 mg cohort due to patient refusing blood draw (1), and due to missed visit (1). mRNA was quantified using Quantigene Plex 2.0 technology according to manufacturer’s protocol (Affymetrix Inc., Santa Clara, CA, USA) and as previously described.13 (link) Probe sets were designed against the human genome for analysis of Nrf2 target genes; a modified version of Panel 11834 (Affymetrix) was used. Genes were normalized to the mean expression of housekeeping genes RPL13A (60S ribosomal protein L13a) and peptidyl-prolyl cis–trans isomerase B.
Peripheral blood cells were quantified by automated cell counter using flow cytometry for white blood cell differential (Sysmex, XN9000).

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Creelan B.C., Gabrilovich D.I., Gray J.E., Williams C.C., Tanvetyanon T., Haura E.B., Weber J.S., Gibney G.T., Markowitz J., Proksch J.W., Reisman S.A., McKee M.D., Chin M.P., Meyer C.J, & Antonia S.J. (2017). Safety, pharmacokinetics, and pharmacodynamics of oral omaveloxolone (RTA 408), a synthetic triterpenoid, in a first-in-human trial of patients with advanced solid tumors. OncoTargets and therapy, 10, 4239-4250.

Publication 2017

Quantigene Plex 2.0 Panel 11834 XN9000

Blood Cell Flow cytometry Genes Housekeeping genes Human genome Mrna Nrf2 Patient Peptidyl prolyl cis trans isomerase b Peripheral blood cells Ribosomal protein Target genes analysis White blood cell

Corresponding Organization :

Other organizations : Moffitt Cancer Center, The Wistar Institute, Georgetown University Medical Center, Reata Pharmaceuticals (United States), AbbVie (United States)

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Variable analysis

independent variables

Dose of treatment (15 mg, 50 mg, 100 mg)

dependent variables

MRNA expression of Nrf2 target genes
Peripheral blood cell counts

control variables

Housekeeping genes (RPL13A and peptidyl-prolyl cis–trans isomerase B) used for normalization of mRNA expression

controls

No positive or negative controls were explicitly mentioned in the input.

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