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Logo of nihpaAbout Author manuscriptsSubmit a manuscriptHHS Public Access; Author Manuscript; Accepted for publication in peer reviewed journal;
Mol Cancer Ther. Author manuscript; available in PMC 2016 July 1.
Published in final edited form as:
PMCID: PMC4497872

Combined Inhibition of Cyclin-Dependent Kinases (Dinaciclib) and AKT (MK-2206) Blocks Pancreatic Tumor Growth and Metastases in Patient-Derived Xenograft Models


KRAS is activated by mutation in the vast majority of cases of pancreatic cancer; unfortunately, therapeutic attempts to inhibit KRAS directly have been unsuccessful. Our previous studies showed that inhibition of cyclin-dependent kinase 5 (CDK5), reduces pancreatic cancer growth and progression, through blockage of the centrally important RAL effector pathway, downstream of KRAS. In the current study, the therapeutic effects of combining the CDK inhibitor dinaciclib (SCH727965; MK-7965) with the pan-AKT inhibitor MK-2206 were evaluated using orthotopic and subcutaneous patient-derived human pancreatic cancer xenograft models. The combination of dinaciclib (20 mg/kg, i.p., t.i.w.) and MK-2206 (60 mg/kg, p.o., t.i.w.) dramatically blocked tumor growth and metastasis in all eight pancreatic cancer models examined. Remarkably, several complete responses were induced by the combination treatment of dinaciclib and MK-2206. The striking results obtained in these models demonstrate that the combination of dinaciclib with the pan-AKT inhibitor MK-2206 is promising for therapeutic evaluation in pancreatic cancer, and strongly suggest that blocking RAL in combination with other effector pathways downstream from KRAS may provide increased efficacy in pancreatic cancer. Based on these data, an NCI-CTEP approved multicenter Phase I clinical trial for pancreatic cancer of the combination of dinaciclib and MK-2206 (NCT01783171) has now been opened.

Keywords: Pancreatic cancer, cyclin-dependent kinase, CDK5, KRAS, patient-derived xenograft


Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related deaths in the United States (1). Despite concerted research efforts, there has been little improvement in PDAC prognosis over the last several decades, and the 5-year survival rate of pancreatic cancer remains below 6% (2). In addition, pancreatic cancer incidence continues to increase; from 2006 to 2010, rates increased by 1.3% per year (1). Therefore, there is an urgent need to find effective systemic therapies to treat this highly lethal cancer.

Activating mutations in KRAS are found in more than 90% of patients with pancreatic cancer (3,4). A series of in vivo evidence shows that mutant KRAS is a driver for tumor initiation and progression in PDAC (59). Thus, oncogenic KRAS is considered a prime therapeutic target for pancreatic cancer. Unfortunately, therapeutic attempts to inhibit mutant KRAS thus far have been unsuccessful (10). A promising alternative strategy has been to target KRAS downstream effector pathways. KRAS has several effector pathways, notably including the PI3K/AKT, RAF/MEK/ERK and RAL effector pathways. Activation of the PI3K/AKT and RAF/MEK/ERK pathways is very common in pancreatic cancer, and these pathways appear to be important to pancreatic cancer growth (6,10,11). Combined inhibition of these pathways has been shown to synergistically inhibit pancreatic cancer growth in preclinical models (1113), and clinical trials to simultaneously inhibit these two pathways are in progress.

Importantly, Counter and colleagues (14,15) have shown that, among the KRAS effector pathways, the RAL pathway is especially critical for the development of pancreatic cancer. This strongly suggests that inhibiting the RAL pathway is a promising central target for blocking dysregulated RAS signaling in pancreatic cancer. However, the RAS/RAL effector pathway has been refractory to inhibition by pharmacological means.

Our previous studies showed that cyclin-dependent kinase 5 (CDK5) is important for RAL activity in pancreatic cancer. CDK5 knockdown, dominant negative expression or treatment with the CDK inhibitor dinaciclib (SCH727965; MK-7965) effectively inhibited RAS/RAL activation and resulted in substantial decreases in cell migration and anchorage independent growth in vitro, and of growth and metastasis of pancreatic cancer xenografts in vivo (16,17). Simultaneous blocking of CDK5 and the PI3K/AKT or RAF/MEK/ERK signaling pathways resulted in further inhibition of anchorage independent growth and cell migration (16). This suggested that such a combination, to inhibit RAL and PI3K/AKT or RAF/MEK/ERK, could be an especially effective therapeutic strategy in pancreatic cancer.

In this study, we show that combining the CDK inhibitor dinaciclib with an inhibitor of the PI3K/AKT pathway, the pan-AKT inhibitor MK-2206, is highly effective in a series of murine orthotopic and subcutaneous patient-derived human pancreatic cancer xenograft models. Based on these data, a Phase I clinical trial has been initiated to evaluate this combination in human pancreatic cancer.

Materials and Methods

Chemicals and reagents

Dinaciclib and MK-2206 were provided by Merck and Co. (Boston, MA). Dinaciclib was dissolved in 20% hydroxypropyl-β-cyclodextrin (HPBCD; Sigma, St. Louis, MO) (18). MK-2206 was dissolved in 0.5% methanol, 0.1% Tween-80.

Generation of orthotopic and subcutaneous xenografts and drug treatment

All small animal experiments described conformed to the guidelines of the Animal Care and Use Committee of Johns Hopkins University. Mice were maintained in accordance with the guidelines of the American Association of Laboratory Animal Care.

Orthotopic xenograft studies

Two modestly gemcitabine sensitive, patient-derived pancreatic cancer xenograft models, Panc265 and Panc253, were chosen to examine the effect of dinaciclib, MK-2206 and dinaciclib + MK-2206 in inhibiting tumor growth and metastases of pancreatic cancer. Low passage subcutaneous xenograft tissue was minced and implanted orthotopically in the pancreas of athymic nude mice, as described in reference 19. Mice were measured by ultrasound (Vevo660, VisualSonics) and randomized by tumor size into 4 treatment groups (n = 7 per group: vehicle control, dinaciclib, MK-2206 and dinaciclib + MK-2206) immediately preceding initiation of therapy (day 14–45 post-implantation). The coefficient of variance among tumor volumes at the start of treatment was ≤10% in each experiment. Treatments were as described above. The total body weights were measured weekly. At the end of the treatment, the mice were euthanized. The primary tumors were harvested and weighed, and the tumor volume were measured with calipers of three orthogonal diameters (a, b and c) and calculated using the formula volume = 1/2(abc). Spleen, pancreas, liver, intestine, colon, lymph node, peritoneum, diaphragm, kidney and lungs were inspected by a thorough necropsy for grossly visible metastases. Since direct, low passage patient derived xenograft (PDX) models were used throughout this study, fluorescent or luminescent assays were not applicable; visual inspection has proven to be accurate for evaluation of metastases in our PDX models (1922).

Subcutaneous xenograft studies

Subcutaneous murine xenografts of low-passage patient-derived human pancreatic cancers were generated as previously described (16,17,23). Six patient-derived xenograft models were chosen at random from the Johns Hopkins PDAC BioBank (23). In each model, tumors were transplanted into both flanks of twenty male CD1 nu/nu athymic mice each. Three to eight weeks after subcutaneous implantation, mice for each case were randomized into four groups of five mice each, and assigned to receive treatment with vehicle control, dinaciclib, MK-2206, or dinaciclib + MK-2206. Total body weights were determined weekly, and xenograft tumor volumes were measured weekly using a digital caliper as previously described (17). After 3–5 weeks of treatment, mice were euthanized and tumor tissues harvested.

Tissue samples were preserved in formalin solution for histology and immunohistochemistry.


For Ki67, cleaved caspase-3, and p-AKT staining of formalin-fixed paraffin-embedded tissue sections, anti-Ki67, or cleaved caspase-3, and p-AKT primary antibody (clone K2, Ventana Medical Systems, Tucson, AZ) was used in combination with a Ventana Benchmark Autostainer as described (16,17). Phospho-Rb (Ser807/811) was stained using a rabbit anti-human polyclonal antibody (#9308, Cell Signaling Technology, Danvers, MA) at a dilution of 1:300 following the standard recommendations provided by the manufacturer. To determine the positive staining cells for Ki67 or the difference in staining intensity for caspase-3, pAkt and pRb, 10 different but histologically similar fields were selected per sample and the slides were analyzed quantitatively using NIH image J software (24). The staining intensity or positive cells measured by the software was plotted using Graph Pad 6.01 (GraphPad Software, Inc.).

Statistical analysis

Two-tailed t tests were performed using Prism version 6.01. P < 0.05 was regarded to be statistically significant. Bar diagrams were generated using Prism version 6.01 and show means and SEMs if not otherwise indicated.


Effective combined treatment with dinaciclib and MK-2206 in orthotopic and subcutaneous patient-derived xenograft models of PDAC

Our previous results showed that CDK5 activity is critical for the RAS/RAL signal transduction pathway in PDAC (16), and that the CDK inhibitor dinaciclib inhibits orthotopic PDAC xenograft growth (17). It has been shown in preclinical studies that inhibition of multiple RAS effector pathways, especially combinations including RAL (25) can be especially effective in limiting growth of RAS-driven human cancers. Therefore, in the current set of experiments, we examined the effect of inhibiting CDK/RAL in combination with another central RAS effector pathway, the PI3K/AKT pathway. We initially chose two highly characterized patient-derived xenograft models of PDAC, Panc253 and Panc265, from the Johns Hopkins PDAC BioBank (23). These KRAS-mutant models closely resemble the pathological conditions of pancreatic cancer in humans. A 2–3 mm3 tumor explant was implanted into the pancreas of nude mice and ultrasound imaging was used to measure the tumor size before randomization and treatment, which began when tumors grew to 50–100 mm3. The combination of dinaciclib (20 mg/kg, i.p., t.i.w.) and MK-2206 (60 mg/kg, p.o., t.i.w.) was well tolerated by the mice, and dramatically blocked tumor growth in the orthotopic Panc265 (90.0%, p<0.001) and Panc253 (93.0%, p<0.001) models (Fig. 1). It also markedly reduced the number of metastatic lesions in both Panc265 (88.2%, p<0.001) and Panc253 (99.0%, p<0.001) tumor models (Fig. 1, Supplementary Fig. S1). Remarkably, complete responses were induced by the combination treatment of dinaciclib and MK-2206 in one mouse in the Panc265 tumor model, and in two mice in the Panc253 tumor model.

Figure 1
Effect of dinaciclib and MK-2206 on orthotopic patient-derived xenograft models

We then examined a larger panel of PDAC patient-derived xenograft models for their sensitivity to the combination of dinaciclib and MK-2206. These six additional models were chosen at random from the Johns Hopkins PDAC BioBank, and examined as subcutaneous xenografts. All of these models have KRAS mutations. The results, shown in Fig. 2, confirm the efficacy of the combination dinaciclib + MK-2206 treatment. Growth of the tumors was significantly reduced in all six models, including several instances of significant tumor regression. Mean growth inhibition in these PDAC models ranged from 63–86%.

Figure 2
Effects of dinaciclib and MK-2206 on 6 additional patient-derived xenograft models of pancreatic cancer

Pharmacodynamic and functional markers

In order to assess the efficacy of dinaciclib and MK-2206 in inhibiting their targets in the PDAC models, we treated the mice 2 hr before euthanasia and tumor harvest (Fig. 3 and Supplementary Fig. 2). For dinaciclib, we examined phospho-RB (pS807/811), a well characterized target of several CDKs (26), and for MK-2206, we examined phospho-AKT (pS473), by Western blotting. Both dinaciclib and MK-2206 efficiently inhibited their respective targets (Fig. 3A and Supplementary Fig. S2). We also examined the ability of dinaciclib and MK-2206 to inhibit cell proliferation, as assessed by immunohistochemistry for Ki67, and to induce apoptosis, as assessed by immunohistochemistry for cleaved caspase 3. Figure 3B and Supplementary Fig. S2 show that either dinaciclib or MK-2206 had a moderate effect in reducing cell proliferation, and that this effect was markedly augmented by combination dinaciclib + MK-2206 treatment. Similarly, apoptosis was induced by each compound, and the combination resulted in increased apoptosis. These results, demonstrating both inhibition of cell proliferation and induction of apoptosis, are consistent with the tumor growth inhibition results shown earlier (Figs. 1 and and22).

Figure 3
Both dinaciclib and MK-2206 are active in xenografts, inhibit proliferation and induce apoptosis


In this study, we explored the effect of a novel combination to target two major effector pathways of RAS signaling, RAL and PI3K, in patient-derived xenograft models of pancreatic cancer. We had shown previously that dinaciclib blocked RAL activation in pancreatic cancer cells, and inhibited human pancreatic cancer xenograft growth in mice (17). Our current study was based on our earlier data indicating that simultaneous inhibition of CDK5 and the PI3K pathway resulted in substantially more effective inhibition of anchorage independent growth of PDAC cells (16). As these earlier studies suggested, the combination of dinaciclib and MK-2206 was quite effective in our patient-derived xenograft models.

Dinaciclib is a potent inhibitor of CDK5 (24), and, in turn, RAL (16,17). Our previous studies showed that CDK5 is a relevant target in PDAC, at least in part through its role in RAL activation (16). However, our studies do not indicate that the sole relevant target of dinaciclib is CDK5/RAL. Like other reported ATP-competitive CDK5 inhibitors (27), dinaciclib inhibits several other CDKs, presumably due in large measure to the homology among the ATP binding pockets of members of the CDK family. Thus, dinaciclib is also a very efficient inhibitor of CDKs 1, 2 and 9 (28); these CDKs have been shown to be significant targets for cancer therapy (29) and may contribute to the antitumor effects we have shown.

Moreover, CDK5 has targets in addition to the RAS/RAL pathway, that are relevant to cancer biology (3032). CDK5 was originally characterized as a neuronal protein involved in cell migration, adhesion and cytoskeletal remodeling (33). These processes are important in cancer progression, and CDK5 has also been shown to be important for these processes in cancer cells, in a cell-specific manner, through phosphorylation of target proteins including FAK, Talin, RAPGEF2, and PIKE-A (3439). CDK5 participates in cell growth and survival, in part by phosphorylation of targets including Rb, NOXA and STAT3 (4045). Effects of CDK5 on secretion, inflammation and angiogenesis are also well characterized (31, 32). These targets are likely to contribute to the overall effect of CDK5 inhibition on the reduced xenograft growth we have observed. While the relative contributions of the various targets of dinaciclib or CDK5 will be difficult to dissect, the contribution of the RAS/RAL pathway may be addressable in future studies, since promising preclinical inhibitors of RAS and RAL have been reported recently (10, 4651).

MK-2206 is an allosteric pan-AKT inhibitor with high specificity for AKT (52). The PI3K/AKT pathway is dysregulated in many cancers and is a well validated therapeutic target. MK-2206, especially in combination, has been shown to have clinical activity against several malignancies (53,54,55), and to sensitize cancer cells to a variety of therapeutic compounds and radiation (56,57). This sensitization may be due in part to the ability of PI3K/AKT activity to augment a large number of survival pathways, including antiapoptosis, DNA repair, and metabolic processes (58,59); such mechanisms may contribute to the significantly improved response to the combination of dinaciclib and MK-2206, compared to either compound alone, that we report here.

Based on the promising results of this study, a multicenter Phase 1 clinical trial (NCT01783171) of a combination of dinaciclib and MK-2206 for pancreatic cancer has been opened.

Supplementary Material


Financial support: This work was supported by grants from NIH-NCI R01 CA134767 (BDN and AM), R01 CA113669 (AM), and P30 CA006973 (Regional Oncology Research Center Cancer Center Support Grant to the Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins).

Dinaciclib and MK-2206 were provided by Merck and Co. Merck also provided unpublished data regarding these compounds, advice on experimental design and interpretation of results.


Conflicts of interest: Rajat Bannerji, Robert Booher, and Peter Strack were employees of Merck and Co. The other authors had no potential conflicts of interest.

Chemical Structures

The chemical structures of dinaciclib and MK-2206 have been published previously (28, 56).


1. Siegel R, Ma J, Zou Z, Jemal A. Cancer statistics, 2014. CA Cancer J Clin. 2014;64:9–29. [PubMed]
2. Von Hoff DD, Ervin T, Arena FP, Chiorean EG, Infante J, Moore M, et al. Increased survival in pancreatic cancer with nab-paclitaxel plus gemcitabine. N Engl J Med. 2013;369:1691–703. [PMC free article] [PubMed]
3. Smit VT, Boot AJ, Smits AM, Fleuren GJ, Cornelisse CJ, Bos JL. KRAS codon 12 mutations occur very frequently in pancreatic adenocarcinomas. Nucleic Acids Res. 1988;16:7773–82. [PMC free article] [PubMed]
4. Almoguera C, Shibata D, Forrester K, Martin J, Arnheim N, Perucho M. Most human carcinomas of the exocrine pancreas contain mutant c-K-ras genes. Cell. 1988;53:549–54. [PubMed]
5. Jones S, Zhang X, Parsons DW, Lin JC, Leary RJ, Angenendt P, et al. Core signaling pathways in human pancreatic cancers revealed by global genomic analyses. Science. 2008;321:1801–6. [PMC free article] [PubMed]
6. di Magliano MP, Logsdon CD. Roles for KRAS in pancreatic tumor development and progression. Gastroenterology. 2013;144:1220–9. [PMC free article] [PubMed]
7. Aguirre AJ, Bardeesy N, Sinha M, Lopez L, Tuveson DA, Horner J, et al. Activated Kras and Ink4a/Arf deficiency cooperate to produce metastatic pancreatic ductal adenocarcinoma. Genes Dev. 2003;17:3112–26. [PubMed]
8. Hingorani SR, Petricoin EF, Maitra A, Rajapakse V, King C, Jacobetz MA, et al. Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse. Cancer Cell. 2003;4:437–50. [PubMed]
9. Collins MA, Bednar F, Zhang Y, Brisset JC, Galbán S, Galbán CJ, et al. Oncogenic Kras is required for both the initiation and maintenance of pancreatic cancer in mice. J Clin Invest. 2012;122:639–53. [PMC free article] [PubMed]
10. Stephen AG, Esposito D, Bagni RK, McCormick F. Dragging ras back in the ring. Cancer Cell. 2014;25:272–81. [PubMed]
11. Hofmann I, Weiss A, Elain G, Schwaederle M, Sterker D, Romanet V, et al. K-RAS mutant pancreatic tumors show higher sensitivity to MEK than to PI3K inhibition in vivo. PLoS One. 2012;7:e44146. [PMC free article] [PubMed]
12. Zhong H, Sanchez C, Spitrzer D, Plambeck-Suess S, Gibbs J, Hawkins WG, et al. Synergistic effects of concurrent blockade of PI3K and MEK pathways in pancreatic cancer preclinical models. PLoS One. 2013;8:e77243. [PMC free article] [PubMed]
13. Alagesan B, Contino G, Guimaraes AR, Corcoran RB, Desphande V, Wojtkiewicz GR, et al. Combined MEK and PI3K inhibition in a mouse model of pancreatic cancer. Clin Cancer Res. 2014 in press. [PMC free article] [PubMed]
14. Lim KH, Counter CM. Reduction in the requirement of oncogenic Ras signaling to activation of PI3K/AKT pathway during tumor maintenance. Cancer Cell. 2005;8:381–92. [PubMed]
15. Lim KH, O’Hayer K, Adam SJ, Kendall SD, Campbell PM, Der CJ, et al. Divergent roles for RalA and RalB in malignant growth of human pancreatic carcinoma cells. Curr Biol. 2006;16:2385–94. [PubMed]
16. Feldmann G, Mishra A, Hong SM, Bisht S, Strock CJ, Ball DW, et al. Inhibiting the cyclin-dependent kinase CDK5 blocks pancreatic cancer formation and progression through the suppression of Ras-Ral signaling. Cancer Res. 2010;70:4460–9. [PMC free article] [PubMed]
17. Feldmann G, Mishra A, Bisht S, Karikari C, Garrido-Laguna I, Rasheed Z, et al. Cyclin-dependent kinase inhibitor Dinaciclib (SCH727965) inhibits pancreatic cancer growth and progression in murine xenograft models. Cancer Biol Ther. 2011;12:598–609. [PMC free article] [PubMed]
18. Booher RN, Hatch H, Dolinski BM, Nguyen T, Harmonay L, Al-Assaad AS, et al. MCL1 and BCL-xL levels in solid tumors are predictive of dinaciclib-induced apoptosis. PLoS One. 2014;9:e108371. [PMC free article] [PubMed]
19. Chenna V, Hu C, Pramanik D, Aftab BT, Karikari C, Campbell NR, et al. A polymeric nanoparticle encapsulated small-molecule inhibitor of Hedgehog signaling (NanoHHI) bypasses secondary mutational resistance to Smoothened antagonists. Mol Cancer Ther. 2012;11:165–73. [PMC free article] [PubMed]
20. Feldmann G, Dhara S, Fendrich V, Bedja D, Beaty R, Mullendore M, et al. Blockade of hedgehog signaling inhibits pancreatic cancer invasion and metastases: a new paradigm for combination therapy in solid cancers. Cancer Res. 2007;67:2187–96. [PMC free article] [PubMed]
21. Bisht S, Mizuma M, Feldmann G, Ottenhof NA, Hong SM, Pramanik D, et al. Systemic administration of polymeric nanoparticle-encapsulated curcumin (NanoCurc) blocks tumor growth and metastases in preclinical models of pancreatic cancer. Mol Cancer Ther. 2010;9:2255–64. [PMC free article] [PubMed]
22. Vincent A, Hong SM, Hu C, Omura N, Young A, Kim H, et al. Epigenetic silencing of EYA2 in pancreatic adenocarcinomas promotes tumor growth. Oncotarget. 2014;5:2575–87. [PMC free article] [PubMed]
23. Rubio-Viqueira B, Jimeno A, Cusatis G, Zhang X, Iacobuzio-Donahue C, Karikari C, et al. An in vivo platform for translational drug development in pancreatic cancer. Clin Cancer Res. 2006;12:4652–61. [PubMed]
24. Collins TJ. ImageJ for microscopy. Biotechniques. 2007;43(1 Suppl):25–30. [PubMed]
25. Hamad NM, Elconin JH, Karnoub AE, Bai W, Rich JN, Abraham RT, et al. Distinct requirements for Ras oncogenesis in human versus mouse cells. Genes Dev. 2002;16:2045–57. [PubMed]
26. Cai D, Byth KF, Shapiro GI. AZ703, an imidazo(1,2-a)pyridine inhibitor of cyclin-dependent kinases 1 and 2, induces E2F-1-dependent apoptosis enhanced by depletion of cyclin-dependent kinase 9. Cancer Res. 2006;66:435–44. [PubMed]
27. Galons H, Oumata N, Gloulou O, Meijer L. Cyclin-dependent kinase inhibitors closer to market launch? Expert Opin Ther Pat. 2013;23:945–63. [PubMed]
28. Parry D, Guzi T, Shanahan F, Davis N, Prabhavalkar D, Wiswell D, et al. Dinaciclib (SCH 727965), a novel and potent cyclin-dependent kinase inhibitor. Mol Cancer Ther. 2010;9:2344–53. [PubMed]
29. Shapiro GI. Cyclin-dependent kinase pathways as targets for cancer treatment. J Clin Oncol. 2006;24:1770–83. [PubMed]
30. Rosales JL, Lee KY. Extraneuronal roles of cyclin-dependent kinase 5. Bioessays. 2006;28:1023–34. [PubMed]
31. Liebl J, Weitensteiner SB, Vereb G, Takács L, Fürst R, Vollmar AM, et al. Cyclin-dependent kinase 5 regulates endothelial cell migration and angiogenesis. J Biol Chem. 2010;285:35932–43. [PMC free article] [PubMed]
32. Liebl J, Fürst R, Vollmar AM, Zahler S. Twice switched at birth: cell cycle-independent roles of the “neuron-specific” cyclin-dependent kinase 5 (Cdk5) in non-neuronal cells. Cell Signal. 2011;23:1698–707. [PubMed]
33. Gupta A, Tsai LH, Wynshaw-Boris A. Life is a journey: a genetic look at neocortical development. Nat Rev Genet. 2002;3:342–55. [PubMed]
34. Xie Z, Sanada K, Samuels BA, Shih H, Tsai LH. Serine 732 phosphorylation of FAK by Cdk5 is important for microtubule organization, nuclear movement, and neuronal migration. Cell. 2003;114:469–82. [PubMed]
35. Strock CJ, Park JI, Nakakura EK, Bova GS, Isaacs JT, Ball DW, et al. Cyclin-dependent kinase 5 activity controls cell motility and metastatic potential of prostate cancer cells. Cancer Res. 2006;66:7509–15. [PubMed]
36. Liu R, Tian B, Gearing M, Hunter S, Ye K, Mao Z. Cdk5-mediated regulation of the PIKE-A-Akt pathway and glioblastoma cell invasion. Proc Natl Acad Sci USA. 2008;105:7570–5. [PubMed]
37. Huang C, Rajfur Z, Yousefi N, Chen Z, Jacobson K, Ginsberg MH. Talin phosphorylation by Cdk5 regulates Smurf1-mediated talin head ubiquitylation and cell migration. Nat Cell Biol. 2009;11:624–30. [PMC free article] [PubMed]
38. Rea K, Sensi M, Anichini A, Canevari S, Tomassetti A. EGFR/MEK/ERK/CDK5-dependent integrin-independent FAK phosphorylated on serine 732 contributes to microtubule depolymerization and mitosis in tumor cells. Cell Death Dis. 2013;4:e815. [PMC free article] [PubMed]
39. Ye T, Ip JP, Fu AK, Ip NY. Cdk5-mediated phosphorylation of RapGEF2 controls neuronal migration in the developing cerebral cortex. Nat Commun. 2014;5:4826. [PMC free article] [PubMed]
40. Fu AK, Fu WY, Ng AK, Chien WW, Ng YP, Wang JH, et al. Cyclin-dependent kinase 5 phosphorylates signal transducer and activator of transcription 3 and regulates its transcriptional activity. Proc Natl Acad Sci USA. 2004;101:6728–33. [PubMed]
41. Lin H, Chen MC, Chiu CY, Song YM, Lin SY. Cdk5 regulates STAT3 activation and cell proliferation in medullary thyroid carcinoma cells. J Biol Chem. 2007;282:2776–84. [PubMed]
42. Lowman XH, McDonnell MA, Kosloske A, Odumade OA, Jenness C, Karim CB, et al. The proapoptotic function of Noxa in human leukemia cellsis regulated by the kinase Cdk5 and by glucose. Mol Cell. 2010;40:823–33. [PubMed]
43. Hsu FN, Chen MC, Lin KC, Peng YT, Li PC, Lin E, et al. Cyclin-dependent kinase 5 modulates STAT3 and androgen receptor activation through phosphorylation of Ser727on STAT3 in prostate cancer cells. Am J Physiol Endocrinol Metab. 2013;305:E975–86. [PubMed]
44. Pozo K, Castro-Rivera E, Tan C, Plattner F, Schwach G, Siegl V, et al. The role of Cdk5 in neuroendocrine thyroid cancer. Cancer Cell. 2013;24:499–511. [PMC free article] [PubMed]
45. Cheung ZH, Gong K, Ip NY. Cyclin-dependent kinase 5 supports neuronal survival through phosphorylation of Bcl-2. J Neurosci. 2008;28:4872–7. [PubMed]
46. Maurer T, Garrenton LS, Oh A, Pitts K, Anderson DJ, Skelton NJ, et al. Small-molecule ligands bind to a distinct pocket in Ras and inhibit SOS-mediated nucleotide exchange activity. Proc Natl Acad Sci U S A. 2012;109:5299–304. [PubMed]
47. Ostrem JM, Peters U, Sos ML, Wells JA, Shokat KM. K-Ras(G12C) inhibitors allosterically control GTP affinity and effector interactions. Nature. 2013;503:548–51. [PMC free article] [PubMed]
48. Yan C, Liu D, Li L, Wempe MF, Guin S, Khanna M, et al. Discovery and characterization of small molecules that target the GTPase Ral. Nature. 2014;515:443–7. [PMC free article] [PubMed]
49. Zimmermann G, Papke B, Ismail S, Vartak N, Chandra A, Hoffmann M, et al. Small molecule inhibition of the KRAS-PDEδ interaction impairs oncogenic KRAS signalling. Nature. 2013;497:638–42. [PubMed]
50. Burns MC, Sun Q, Daniels RN, Camper D, Kennedy JP, Phan J, et al. Approach for targeting Ras with small molecules that activate SOS-mediated nucleotide exchange. Proc Natl Acad Sci USA. 2014;111:3401–6. [PubMed]
51. Lim SM, Westover KD, Ficarro SB, Harrison RA, Choi HG, Pacold ME, et al. Therapeutic targeting of oncogenic K-Ras by a covalent catalytic site inhibitor. Angew Chem Int Ed Engl. 2014;53:199–204. [PMC free article] [PubMed]
52. Yan L. MK-2206: a potent oral allosteric AKT inhibitor. AACR Annual Meeting; 2009; p. Abstract Number: DDT01–1.
53. Yap TA, Yan L, Patnaik A, Fearen I, Olmos D, Papadopoulos K, et al. First-in-man clinical trial of the oral pan-AKT inhibitor MK-2206 in patients with advanced solid tumors. J Clin Oncol. 2011;29:4688–95. [PubMed]
54. Tolcher AW, Khan K, Ong M, Banerji U, Papadimitrakopoulou V, Gandara DR, et al. Antitumor Activity in RAS-Driven Tumors by Blocking AKT and MEK. Clin Cancer Res. 2015;21:739–48. [PMC free article] [PubMed]
55. Gupta S, Munster PN, Hollebecque A, Argiles G, Dajani O, Cheng JD, et al. Safety/efficacy of MK-8669 (ridaforolimus) plus MK-2206 (AKT inhibitor) in patients with advanced breast cancer with low RAS signature and PTEN deficient prostate cancer. J Clin Oncol. 2014;32(suppl):5s. abstr 2509.
56. Hirai H, Sootome H, Nakatsuru Y, Miyama K, Taguchi S, Tsujioka K, et al. MK-2206, an allosteric Akt inhibitor, enhances antitumor efficacy by standard chemotherapeutic agents or molecular targeted drugs in vitro and in vivo. Mol Cancer Ther. 2010;9:1956–67. [PubMed]
57. Stegeman H, Kaanders JH, Wheeler DL, van der Kogel AJ, Verheijen MM, Waaijer SJ, et al. Activation of AKT by hypoxia: a potential target for hypoxic tumors of the head and neck. BMC Cancer. 2012;12:463. [PMC free article] [PubMed]
58. Brown KK, Toker A. The phosphoinositide 3-kinase pathway and therapy resistance in cancer. F1000Prime Rep. 2015;7:13. [PMC free article] [PubMed]
59. Cassinelli G, Zuco V, Gatti L, Lanzi C, Zaffaroni N, Colombo D, et al. Targeting the Akt kinase to modulate survival, invasiveness and drug resistance of cancer cells. Curr Med Chem. 2013;20:1923–45. [PubMed]