Protein kinase D (PKD) family members are increasingly implicated in multiple normal and abnormal
biological functions, including signaling pathways that promote mitogenesis in
pancreatic cancer. However, nothing is known about the effects of targeting PKD in
pancreatic cancer. Our PKD inhibitor discovery program identified
CRT0066101 as a specific inhibitor of all PKD
isoforms. The aim of our study was to determine the effects of
CRT0066101 in
pancreatic cancer. Initially, we showed that autophosphorylated PKD1 and PKD2 (activated PKD1/2) are significantly upregulated in
pancreatic cancer and that PKD1/2 are expressed in multiple
pancreatic cancer cell lines. Using Panc-1 as a model system, we showed that
CRT0066101 reduced
bromodeoxyuridine incorporation; increased apoptosis; blocked
neurotensin-induced PKD1/2 activation; reduced
neurotensin-induced, PKD-mediated Hsp27 phosphorylation; attenuated PKD1-mediated
NF-kappaB activation; and abrogated the expression of
NF-kappaB-dependent proliferative and prosurvival
proteins. We showed that
CRT0066101 given orally (80 mg/kg/d) for 24 days significantly abrogated
pancreatic cancer growth in Panc-1 subcutaneous xenograft model. Activated PKD1/2 expression in the treated
tumor explants was significantly inhibited with peak
tumor concentration (12 micromol/L) of
CRT0066101 achieved within 2 hours after
oral administration. Further, we showed that
CRT0066101 given orally (80 mg/kg/d) for 21 days in Panc-1 orthotopic model potently blocked
tumor growth in vivo.
CRT0066101 significantly reduced Ki-67-positive proliferation index (P < 0.01), increased
terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive apoptotic cells (P < 0.05), and abrogated the expression of
NF-kappaB-dependent
proteins including
cyclin D1,
survivin, and cIAP-1. Our results show for the first time that a PKD-specific small-molecule inhibitor
CRT0066101 blocks
pancreatic cancer growth in vivo and show that PKD is a novel therapeutic target in
pancreatic cancer.