Activation of Grm1 expression by mutated BRaf (V600E) in vitro and in vivo

// Ho-Chung Chen 1, * , Jairo Sierra 1, 2, * , Lumeng Jenny Yu 1, * , Robert Cerchio Jr. 1, 3 , Brian A. Wall 1, 4 , James Goydos 2, 5 and Suzie Chen 1, 2, 3, 5 1 Susan Lehman Cullman Laboratory for Cancer Research, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway 08854, NJ, USA 2 Rutgers-GSBS at Robert Wood Johnson Medical School, Piscataway 08854, NJ, USA 3 Pharmacology and Toxicology Graduate Program, Rutgers University, Piscataway 08854, NJ, USA 4 Global Product Safety, Colgate-Palmolive Company, Piscataway 08854, NJ, USA 5 Rutgers Cancer Institute of New Jersey, New Brunswick 08903, NJ, USA * These authors contributed equally to this work Correspondence to: Suzie Chen, email: suziec@pharmacy.rutgers.edu Keywords: mutated BRaf (V600E); Grm1; melanoma; senescence; p15 Received: September 27, 2017      Accepted: December 08, 2017      Published: December 23, 2017 ABSTRACT Our laboratory previously showed that ectopic expression of Grm1 is sufficient to induce spontaneous melanoma formation with 100% penetrance in transgenic mouse model, TG-3, which harbors wild-type BRaf. Studies identified Grm1 expression in human melanoma cell lines and primary to secondary metastatic melanoma biopsies having wild-type or mutated BRaf, but not in normal melanocytes or benign nevi. Grm1 expression was detected in tissues from mice genetically engineered with inducible melanocyte-specific BRaf V600E . Additionally, stable clones derived from introduction of exogenous BRaf V600E in mouse melanocytes also showed Grm1 expression, which was not detected in the parental or empty vector-derived cells, suggesting that expression of BRaf V600E could activate Grm1 expression. Despite aberrant Grm1 expression in the inducible, melanocyte-specific BRaf V600E mice, no tumors formed. However, in older mice, the melanocytes underwent senescence, as demonstrated previously by others. It was proposed that upregulated p15 and TGFβ contributed to the senescence phenotype. In contrast, in older TG-3 mice the levels of p15 and TGFβ remained the same or lower. Taken together, these results suggest the temporal regulation on the expression of “oncogenes” such as Grm1 or BRaf V600E is critical in the future fate of the cells. If BRaf V600E is turned on first, Grm1 expression can be induced, but this is not sufficient to result in development of melanoma; the cells undergo senescence. In contrast, if ectopic expression of Grm1 is turned on first, then regardless of wild-type or mutated BRaf in the melanocytes melanoma development is the consequence.