Figure 4 from TNG961 Is a Selective Oral HBS1L Molecular Glue Degrader for the Treatment of FOCAD-Deleted Cancers

Abstract

TNG961 induces potent, selective CRBN-dependent degradation of HBS1L. A, Endogenous HBS1L degradation and PELO destabilization measured by Western blot after 24 hours of treatment with TNG961 in the indicated cell lines. The top dose was 10 µmol/L, tested as a 9-point, 3-fold dilution series. Representative images are from N = 2 biological replicates. B, CRBN dependence of HBS1L degradation and PELO destabilization assessed by Western blot after 24 hours of treatment with the indicated concentrations of TNG961 in parental and CRBN knockout (sgCRBN) MIAPACA2 cells. Representative images are from N = 2 biological replicates. C, Neddylation dependence assessed by Western blot after 24 hours of treatment with TNG961 alone or in combination with 100 nmol/L MLN4924 in MIAPACA2 cells. Representative images are from N = 2 biological replicates. D, Degron-loop dependence assessed by Western blot after 24 hours of treatment with the indicated concentrations of TNG961 in parental MIAPACA2 cells and in engineered lines expressing either HBS1L G625N (covering endogenous HBS1L knockout) or GSPT1 G575N (covering endogenous GSPT1 knockout). Representative images are from N = 3 biological replicates. E, Global quantitative proteomics in MM.1s cells after 6 hours of treatment with 500 nmol/L TNG961 or DMSO. Data are shown as relative protein abundance (TNG961/DMSO); HBS1L, PELO, and common IMiD neosubstrates are annotated. F, Western blot analysis of common CRBN neosubstrates after 24 hours of treatment with TNG961 at the indicated concentrations. Representative images are from N = 2 biological replicates.