Figure S1 - Sensitivity of NB cell lines to allosteric SHP2 inhibitors. Figure S2 - Sensitivity of NB cell lines to catalytic SHP2 inhibitors. Figure S3 - Sensitivity to SHP2 inhibitors in isogenic NB cells with exogenous PTPN11 mutations. Figure S4 - NRAS status determines sensitivity to SHP2 and MAPK inhibition. Figure S5 - MAPK inhibitors sensitize NB cells to SHP099 and II-B08. Figure S6 - SHP2 inhibitors are synergistic with trametinib and other MAPK inhibitors. Figure S7 - MAPK inhibitors synergize with SHP2 inhibitors in ALK-mutant NB cells. Figure S8 - Assessment of drug tolerability for SHP099 and trametinib in vivo. Figure S9 - Pre-clinical assessment of SHP099 plus trametinib in NRAS-mutant NB xenografts. Figure S10 - Pre-clinical assessment of SHP099 plus ulixertinib in NRAS-mutant NB xenografts.
Funding
Canadian Institutes of Health Research and James
Canadian Institutes of Health Research
Sick Kids Neuroblastoma Research
James Fund
Lilah's Fund
Sebastian's Superheroes
ARTICLE ABSTRACT
Survival for high-risk neuroblastoma remains poor and treatment for relapsed disease rarely leads to long-term cures. Large sequencing studies of neuroblastoma tumors from diagnosis have not identified common targetable driver mutations other than the 10% of tumors that harbor mutations in the anaplastic lymphoma kinase (ALK) gene. However, at neuroblastoma recurrence, more frequent mutations in genes in the RAS–MAPK pathway have been detected. The PTPN11-encoded tyrosine phosphatase SHP2 is an activator of the RAS pathway, and we and others have shown that pharmacologic inhibition of SHP2 suppresses the growth of various tumor types harboring KRAS mutations such as pancreatic and lung cancers. Here we report inhibition of growth and downstream RAS–MAPK signaling in neuroblastoma cells in response to treatment with the SHP2 inhibitors SHP099, II-B08, and RMC-4550. However, neuroblastoma cell lines harboring endogenous NRASQ61K mutation (which is commonly detected at relapse) or isogenic neuroblastoma cells engineered to overexpress NRASQ61K were distinctly resistant to SHP2 inhibitors. Combinations of SHP2 inhibitors with other RAS pathway inhibitors such as trametinib, vemurafenib, and ulixertinib were synergistic and reversed resistance to SHP2 inhibition in neuroblastoma in vitro and in vivo. These results suggest for the first time that combination therapies targeting SHP2 and other components of the RAS–MAPK pathway may be effective against conventional therapy-resistant relapsed neuroblastoma, including those that have acquired NRAS mutations.
These findings suggest that conventional therapy–resistant, relapsed neuroblastoma may be effectively treated via combined inhibition of SHP2 and MEK or ERK of the RAS–MAPK pathway.
