Open AccessNHD2-15, a novel antagonist of Growth Factor Receptor-Bound Protein-2 (GRB2), inhibits leukemic proliferation
Author(s) -
Tina R. Lewis,
Jesse Smith,
Kallie Griffin,
Stephanie Aguiar,
Kristen F. Rueb,
Natalie HolmbergDouglas,
Ellen M. Sampson,
Skylar Tomasetti,
Sofía Rodríguez,
David L. Stachura,
Carolynn C. Arpin
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0236839
Subject(s) - grb2 , biology , fusion protein , breakpoint cluster region , cancer research , microbiology and biotechnology , myeloid leukemia , k562 cells , cell growth , sh3 domain , abl , signal transduction , receptor , chemistry , leukemia , proto oncogene tyrosine protein kinase src , biochemistry , immunology , tyrosine kinase , gene , recombinant dna
The majority of chronic myeloid leukemia (CML) cases are caused by a chromosomal translocation linking the breakpoint cluster region (BCR) gene to the Abelson murine leukemia viral oncogene-1 (ABL1), creating the mutant fusion protein BCR-ABL1. Downstream of BCR-ABL1 is growth factor receptor-bound protein-2 (GRB2), an intracellular adapter protein that binds to BCR-ABL1 via its src-homology-2 (SH2) domain. This binding constitutively activates growth pathways, downregulates apoptosis, and leads to an over proliferation of immature and dysfunctional myeloid cells. Utilizing novel synthetic methods, we developed four furo-quinoxaline compounds as GRB2 SH2 domain antagonists with the goal of disrupting this leukemogenic signaling. One of the four antagonists, NHD2-15, showed a significant reduction in proliferation of K562 cells, a human BCR-ABL1 + leukemic cell line. To elucidate the mode of action of these compounds, various biophysical, in vitro , and in vivo assays were performed. Surface plasmon resonance (SPR) assays indicated that NHD2-15 antagonized GRB2, binding with a K D value of 119 ± 2 μM. Cellulose nitrate (CN) assays indicated that the compound selectively bound the SH2 domain of GRB2. Western blot assays suggested the antagonist downregulated proteins involved in leukemic transformation. Finally, NHD2-15 was nontoxic to primary cells and adult zebrafish, indicating that it may be an effective clinical treatment for CML.