| Autor: |
Booth L; a Departments of Biochemistry and Molecular Biology , Virginia Commonwealth University , Richmond , VA , USA., Roberts JL; a Departments of Biochemistry and Molecular Biology , Virginia Commonwealth University , Richmond , VA , USA., Rais R; a Departments of Biochemistry and Molecular Biology , Virginia Commonwealth University , Richmond , VA , USA., Cutler RE Jr; c Puma Biotechnology Inc ., Los Angeles , CA , USA., Diala I; c Puma Biotechnology Inc ., Los Angeles , CA , USA., Lalani AS; c Puma Biotechnology Inc ., Los Angeles , CA , USA., Poklepovic A; b Medicine , Virginia Commonwealth University , Richmond , VA , USA., Dent P; a Departments of Biochemistry and Molecular Biology , Virginia Commonwealth University , Richmond , VA , USA. |
| Abstrakt: |
Cancers expressing mutant RAS are associated with a weaker response to chemotherapy and a shorter overall patient survival. We have demonstrated that the irreversible inhibitor of ERBB1/2/4, neratinib, inhibits ERBB1/2/4 and causes their internalization and autolysosomal degradation. Fellow-traveler membrane proteins with RTKs, including mutant K-/N-RAS, were also degraded. We discovered that the CDK4/6 inhibitor palbociclib increased autophagosome and then autolysosome levels in a time dependent fashion, did not reduce mTOR activity, and interacted with temsirolimus to kill. Neratinib and palbociclib interacted in a greater than additive manner to increase autophagosome and then autolysosome levels in a time dependent fashion, and to cause tumor cell killing. Killing required the expression of ATM and AMPKα, Beclin1 and ATG5, BAX and BAK and of AIF, but not of caspase 9. In some cells over-expression of BCL-XL was protective whereas in others it was ineffective. The lethality of [neratinib + palbociclib] was modestly enhanced by the PDE5 inhibitor sildenafil and strongly enhanced by the HDAC inhibitor sodium valproate. This was associated with K-RAS degradation and a greater than additive increase in autophagosome and autolysosome levels. Killing by the three-drug combination required ATM and AMPKα, and, to a greater extent, Beclin1 and ATG5. In vivo, [valproate + palbociclib] and [neratinib + valproate + palbociclib] interacted to suppress the growth of a carboplatin/paclitaxel resistant PDX ovarian tumors that express a mutant N-RAS. Our data support performing a future three-drug trial with these agents. |
