Abstract A096: RAS mutation status predicts activity of the SHP2 inhibitor TNO155 in RAS-pathway driven fusion-negative rhabdomyosarcoma

Abstract

Abstract Rhabdomyosarcomas (RMS) are the most common pediatric soft tissue sarcoma. Fusion-negative RMS (FN-RMS, the subset lacking FOXO1 translocations), are characterized by oncogenic point mutations in genes that regulate activity of the RAS/ERK signaling pathway. Clinically available agents that directly target oncogenic RAS in cancer have only recently become FDA approved, and their activity is limited to cancers with G12C mutations in KRAS. Alternate molecular strategies to target RAS activation in cancer include inhibitors of RAS regulatory proteins, including the nonreceptor tyrosine phosphatase SHP2. SHP2 regulates RAS signaling by dephosphorylating receptor tyrosine kinases (RTKs), preventing effective RAS GTPase-activating protein (GAP) recruitment. Additionally, SHP2 recruits SOS1, a RAS guanine nucleotide exchange factor (GEF), promoting RAS activation. Novel small molecule inhibitors of SHP2 are now in clinical trials in several cancer types. We set out to determine whether SHP2 inhibitors demonstrate preclinical activity in models of RAS-pathway driven FN-RMS. We hypothesize that specific oncogenic mutations in HRAS, NRAS and KRAS, affecting codons 12, 13 and 61, will determine SHP2 inhibitor sensitivity in models of FN-RMS, and that sensitivity will be determined based on the interactions between RAS and RAS-GAPs/RAS-GEFs. We tested the effects of SHP2 inhibition on ERK pathway signaling, using the small molecule inhibitor TNO155, in a panel of FN-RMS cell lines with varied RAS mutation status. In FN-RMS cells with Q61-mutant HRAS or NRAS, phospho-MEK and phospho-ERK were unchanged, while in cells with RTK-amplification and wild-type RAS, or those with G12 or G13-mutant HRAS we observed reductions in p-MEK and p-ERK following treatment with TNO155. Reduced 2-D and 3-D cell growth were also observed in WT and G12/G13-mutant HRAS cell lines, but not those with Q61 mutations, using both high throughput proliferation assays and soft-agar colony formation assays, respectively. Responses in sensitive cells were modest and cytostatic at best, suggesting single agent SHP2i may have insufficient activity in these tumors. RNAseq analysis of TNO155-sensitive RAS-WT RMS cells reveals a set of differentially expressed genes that may inform the selection of agents for combination therapy. In addition, a subset of FN-RMS cells also harbor activating mutations in FGFR and PI3K-associated genes, and we therefore tested whether TNO155 might synergize with FGFR or PI3K small-molecule inhibitors and found additive effects between TNO155, the PI3Ki alpelisib, and the pan-FGFRi erdafitinib. In conclusion, FN-RMS cells driven by RTK-amplification, or by RAS G12 or G13 mutations, are sensitive to TNO155 while cells driven by RAS Q61 mutations are not. Additionally, FN-RMS cells treated with TNO155 may rely on the PI3K pathway for adaptive resistance, and combination treatment involving TNO155 and FGFR or PI3K inhibitors may reduce tumor growth with better efficacy than either compound alone. Citation Format: Andrew Baker, Patience Odeniyide, Alyza Skaist, Alla Lisok, Elana Fertig, Jiawan Wang, Christine Pratilas. RAS mutation status predicts activity of the SHP2 inhibitor TNO155 in RAS-pathway driven fusion-negative rhabdomyosarcoma [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A096.