Targeting CDK4/6 and the Cell Cycle in Combination with Bortezomib in the 5T33MM Myeloma Model.

Multiple myeloma (MM) is ultimately fatal in part because no effective cell cycle-based therapy has been available to control tumor cell proliferation. Loss of cell cycle control in MM cells stems from coordinated deregulation of Cdk4-cyclin D1 or Cdk6 (Cdk4)-cyclin D2. PD 0332991 is the first orally bioactive small molecule that potently and specifically inhibits Cdk4 and Cdk6. It represents a promising cell cycle-based therapy for myeloma owing to its ability to potently inhibit Cdk4/6 and induce G1 cell cycle arrest in primary human myeloma cells in BM stromal cell co-cultures and to control tumor progression in a xenograft model. However, the efficacy of PD 0332991 in the presence of an intact immune system is unknown. To optimize therapeutic targeting of Cdk4/6 with PD 0332991, we investigated the effectiveness of PD 0332991 in inhibiting Cdk4/6 and controlling myeloma tumor progression in the immunocompetent, bone migrating 5T33MM model. By quantitative real-time PCR analysis, we found that these myeloma cells express a normal plasma cell transcription program such as upregulated Blimp-1 and loss of Bcl-6 expression. However, they proliferate aggressively due to Cdk4 overexpression and impaired p27Kip1 expression, thereby mimicking relapse disease in human myeloma. PD 0332991 potently inhibits Cdk4/6 phosphorylation of Rb in primary 5T33MM cells and induces G1 cell cycle arrest, both in vivo and ex vivo. Accordingly, treatment with PD 0332991 significantly prolongs the survival of tumor-induced 5T33MM mice; a mean of 35 days in the PD 0332991-treated group (N=9) versus 25 days in the vehicle-treated group (N=9, p< 0.003). These findings demonstrate for the first time that PD 0332991 targets Cdk4/6 and controls tumor expansion in the presence of an intact immune system. To further optimize Cdk4/6 targeting in MM, we combined PD 0332991 with bortezomib, a cytotoxic drug widely used in MM treatment. Pretreatment of 5T33MMvt cells with PD 0332991 for 48 hours markedly augmented bortezomib killing, to the same level of twice the dose of bortezomib when it was used alone. These studies of PD 0332991 in the immunocompetent 5T33MM model demonstrate that, as a consequence of Cdk4/6 inhibition and induction of G1 arrest, PD 0332991 sensitizes MM cells to killing by a cytotoxic agent. Targeting Cdk4/6 in combination therapy, therefore, represents a novel and promising strategy for myeloma treatment.