Targeting the epichaperome as an effective precision medicine approach in a novel

摘要

a Hematoxylin and eosin (H&E) staining of the bone marrow (BM) biopsy showing hypercellular BM with unclassified myeloproliferative disorder at disease onset. Hematopathology evaluation from 45 months prior this study. b BM aspirate shows increased myeloblasts as patient progressed to acute myeloid leukemia (AML). Hematopathology evaluation 17 months prior to this study. c FISH analysis for SYK showing break apart (rearranged) green and red signals, and yellow (unrearranged) allele (for PML FISH data see Supplementary Fig. 1). d Representative gating of leukocyte subpopulations evaluated with flow cytometry. Blasts, lymphocytes and granulocytes were sorted by gating on SSC-A vs. CD45 from bone marrow (BM) sample at baseline. e Agarose gel electrophoresis of PCR amplicon showing positive PML-SYK fusion gene transcripts in bulk population and sorted populations indicated in d. f Schematic representation of cells with high (top row) or no epichaperome (bottom row) with corresponding expected flow histograms in PU-FITC biding assay. PU-FITC binds to intracellular epichaperome with high affinity and gives signal indicating abundance of epichaperome. g Preclinical evaluation of epichaperome abundance in lymphocytes (green), granulocytes (blue) and blasts (red) by performing PU-FITC-binding assay. The intensity of FITC fluorescence indicates the level of epichaperome and unstained control is shown in gray. h and i Viability assay with cells from WCM254 (red), MV4;11 (blue, control AML cell line with high epichaperome and with high sensitivity to PU-H71) or normal peripheral blood (nPB) mononuclear cells (black) treated with escalating doses of PU-H71 for 72 h in vitro. Representative flow plots (h) and graphic representation at increasing doses of PU-H71 (i). Experiments performed in triplicate, showing the mean and error bar represents the SD.