Clinical significance of genomic alterations of the CDK4-pathway and sensitivity to the CDK4 inhibitor PD 0332991 in melanoma.

Abstract

8520 Background: Activation of CDK4 by amplification, increased expression of Cyclin D1 (CCND1) or reduced expression of the CDK inhibitor p16 (CDKN2A) can contribute to transformation of melanocytes indicating that CDK4 can act as an oncogene in melanoma.To explore if CDK4 may be a viable target for the treatment of human melanoma we have analyzed the frequency and clinico-pathological associations of genomic alterations of the CDK4 pathway in primary human melanoma and examined the genomic predictors of sensitivity to the highly selective CDK4/6 inhibitor PD 0332991 (991) in a panel of melanoma cell lines. Methods: A series of 167 primary melanomas with clinical, molecular and pathological annotation, including median follow up of 6.6 years, were analyzed for copy number variation (CNV)- gain of CDK4 or CCND1 (average gene copy >2.4) or loss of CDKN2A (average gene copy <1.4), by fluorescence in situ hybridization. A panel of 39 cell lines were treated with 991 in vitro and GI50s calculated. The mean GI50 of the melanoma cell lines was used to define sensitivity. Gene expression profiling and mutation or CNV in CDKN2A were used to identify predictors of sensitivity. Results: 75% of primary melanomas had at least one CNV (75%, 70% and 82% for BRAF, NRAS or wild-type BRAF/NRAS mutation status respectively). 55% showed loss of CDKN2A. 28% of melanomas had two or more CNVs. Melanomas with two or more CNVs involving CCND1 had worse overall survival (HR 5.56, p=0.02). Low CDKN2A mRNA expression or mutation or loss of CDKN2A predicted sensitivity to 991 with 30/33 mutant/loss lines being sensitive compared to only 2/6 wild type lines (p<0.006). Expression of CDK4, CCND1 or other cyclins or CDK-inhibitors did not predict sensitivity to 991. Conclusions: Genomic alterations in the CDK4 pathway are frequent in melanoma and are associated with worse survival, particularly when melanomas harbor two or more CNVs involving CCND1. Mutation, loss or low expression of CDKN2A in melanoma cell lines predicted sensitivity to the CDK4 inhibitor 991. Taken together these data support evaluation of CDK4 inhibitors in melanoma and suggest that CDKN2A maybe a genomic predictor of sensitivity to these agents.

Kinase inhibitor screening identifies CDK4 as a potential therapeutic target for melanoma

“…Notably, WM-266-4 cells were significantly more sensitive to growth inhibition by fascaplysin than WM-115 cells. WM-115 is a primary melanoma cell line, while WM-266-4 is a metastatic cell line, derived from the same patient ( 25 ). The different sensitivity may be related to the faster proliferation rate of the metastatic cell line WM-266-4 ( 26 ).…”

Achievements and Challenges of Molecular Targeted Therapy in Melanoma

“…38,39 Other mechanisms of primary resistance include genetic aberrations that lead to cell cycle activation (including cyclin D amplifıcation/expression, p16 loss, and CDK4 activation) or parallel growth factor signaling (PTEN loss). [40][41][42]…”

Major therapeutic developments and current challenges in advanced melanoma

“…In addition to driving oncogenes, a majority of melanomas either have aberrations in genes involved in cell cycle regulation [including loss of the cyclin-dependent kinase (CDK) inhibitor p16 (CDKN2A) or gain of function of cyclin D1 (CCND1), CDK4 or CDK6] and/or phosphatase and tensin homolog (PTEN), a key negative regulator of the phosphoinositol-3-kinase (PI3K) pathway. [29][30][31] As the molecular drivers of melanoma were being identified, small-molecule inhibitors were being designed and developed to block the aberrant signalling associated with these oncogenic mutations. Figure 2 highlights the major signalling cascades described in melanoma oncogenesis, namely the PI3K and MAPK pathways.…”

“…For example, BRAF mutations are present in over 50% of melanomas that arise in nonchronic sun‐damaged skin but approximately only 10% of those that arise on chronic sun‐damaged skin, or acral or mucosal surfaces. In addition to driving oncogenes, a majority of melanomas either have aberrations in genes involved in cell cycle regulation [including loss of the cyclin‐dependent kinase (CDK) inhibitor p16 (CDKN2A) or gain of function of cyclin D1 (CCND1), CDK4 or CDK6] and/or phosphatase and tensin homolog (PTEN), a key negative regulator of the phosphoinositol‐3‐kinase (PI3K) pathway …”