Shh signaling and pancreatic cancer: implications for therapy

Hedgehog signaling has been implicated in the development of several human cancers, including small cell lung carcinomas, medulloblastomas, basal cell carcinomas, and digestive tract tumors. Elevated levels of pathway components are observed in pancreatic ductal adenocarcinoma (PDAC) precursor lesions, and these levels increase further as lesions progress to more advanced stages. Yet the mechanisms by which hedgehog signaling contributes to pancreatic tumorigenesis were poorly understood. We recently published results showing that activated hedgehog signaling enhances the proliferation and survival of pancreatic duct epithelial cells, the presumptive target cells for PDAC development. We also demonstrated that sonic hedgehog (Shh) expression, in cooperation with loss of the Trp53 and Ink4a/Arf tumor suppressor loci, was sufficient to initiate the formation of early pancreatic lesions. Furthermore, Shh signaling enhanced K-Ras-mediated pancreatic tumorigenesis and reduced the dependence of tumor cells on the sustained activation of Ras-stimulated signaling pathways. Here we discuss the significance of these findings and the implications for therapy

MiR-142-3p is downregulated in aggressive p53 mutant mouse models of pancreatic ductal adenocarcinoma by hypermethylation of its locus

Pancreatic ductal adenocarcinoma (PDAC) is an extremely aggressive disease with poor prognostic implications. This is partly due to a large proportion of PDACs carrying mutations in TP53, which impart gain-of-function characteristics that promote metastasis. There is evidence that microRNAs (miRNAs) may play a role in both gain-of-function TP53 mutations and metastasis, but this has not been fully explored in PDAC. Here we set out to identify miRNAs which are specifically dysregulated in metastatic PDAC. To achieve this, we utilised established mouse models of PDAC to profile miRNA expression in primary tumours expressing the metastasis-inducing mutant p53R172H and compared these to two control models carrying mutations, which promote tumour progression but do not induce metastasis. We show that a subset of miRNAs are dysregulated in mouse PDAC tumour tissues expressing mutant p53R172H, primary cell lines derived from mice with the same mutations and in TP53 null cells with ectopic expression of the orthologous human mutation, p53R175H. Specifically, miR-142-3p is downregulated in all of these experimental models. We found that DNA methyltransferase 1 (Dnmt1) is upregulated in tumour tissue and cell lines, which express p53R172H. Inhibition or depletion of Dnmt1 restores miR-142-3p expression. Overexpression of miR-142-3p attenuates the invasive capacity of p53R172H-expressing tumour cells. MiR-142-3p dysregulation is known to be associated with cancer progression, metastasis and the miRNA is downregulated in patients with PDAC. Here we link TP53 gain-of-function mutations to Dnmt1 expression and in turn miR-142-3p expression. Additionally, we show a correlation between expression of these genes and patient survival, suggesting that they may have potential to be therapeutic targets

The role of Fas signalling and the c-MYC oncogene in T cell apoptosis and transformation

Cancer is a disease characterised by disruption of apoptotic pathways and inappropriate expression of survival or proliferative signals. Despite its role as a major apoptotic pathway in T cells however, the role of Fas in lymphomagenesis is not clear. To investigate the role of Fas in MYC induced lymphomagenesis, animals harbouring a c-MYC transgene, with expression targeted to the T cell lineage, were placed on a Faslpr background. Loss of Fas did not alter the incidence, latency or phenotype of thymic lymphomas arising in these mice. In addition, the incidence and latency of lymphomas in Faslpr mice infected with MuLV was not significantly different from strain controlled mice. Further, the proportion of lymphomas with retroviral insertions at c-myc was not increased in Faslpr mice. These results indicate that Fas does not act to restrict tumourigenesis, at least in the T cell lineage. Previous studies have reported that MYC induced apoptosis can occur through Fas and p53 signalling pathways. However loss of Fas did not inhibit MYC induced apoptosis in normal or neoplastic T cells, indicating that MYC induced apoptosis can occur by a Fas independent pathway. Furthermore, MYC induced apoptosis could occur in the combined absence of both Fas and p53 apoptotic pathways. Although loss of these two major apoptotic pathways did not prevent MYC induced apoptosis, protection from MYC induced apoptosis was observed with cell contact in some cell lines. This protection was shown to dependent on a PI3 kinase pathway. In addition, in cell lines that retained functional p53, the PI3 kinase/Akt survival signal was shown to be critical for cell survival. The response to T cell receptor activation in Faslpr thymocytes was also examined. In addition to lacking the ability to undergo activation induced cell death, a proliferative defect was revealed in Faslpr thymocytes, compared with control MRL thymocytes. This defect was rescued by co-stimulation of the cell surface marker CD28. The role of Fas:FasL interactions in tumourigenesis may depend on the cell types in which Fas and FasL are expressed. Signalling through Fas or FasL may be important in the transduction of proliferative signals in T cells, and this may explain why disabling the Fas pathway does not appear to influence T cell lymphomagenesis

Neutrophils: homing in on the myeloid mechanisms of metastasis

The metastasis cascade is complex and comprises several stages including local invasion into surrounding tissue, intravasation and survival of tumour cells in the circulation, and extravasation and colonisation of a distant site. It is increasingly clear that these processes are driven not only by signals within the tumour cells, but are also profoundly influenced by stromal cells and signals in the tumour microenvironment. Amongst the many cell types within the tumour microenvironment, immune cells such as lymphocytes, macrophages and neutrophils play a prominent role in tumour development and progression. Neutrophils, however, have only recently emerged as important players, particularly in metastasis. Here we review the current evidence suggesting a multi-faceted role for neutrophils in the metastatic cascade

RNA polymerase III transcription is repressed in response to the tumour suppressor ARF

The tumour suppressor protein ARF provides a defence mechanism against hyperproliferative stresses that can result from the aberrant activation of oncogenes. Accordingly, ARF is silenced or deleted in many human cancers. Activation of ARF can arrest growth and cell cycle progression, or trigger apoptosis. A principle mediator of these effects is p53, which ARF stabilizes by binding and inhibiting MDM2. However, ARF has additional targets and remains able to block growth in the absence of p53, albeit less efficiently. For example, ARF can suppress rRNA production in a p53-independent manner. We have found that the synthesis of tRNA by RNA polymerase III is also inhibited in response to ARF. However, in contrast to its effects on rRNA synthesis, ARF is unable to inhibit tRNA gene transcription when p53 is ablated. These results add to the growing list of cellular changes that can be triggered by ARF induction

Sheep recognize familiar and unfamiliar human faces from two-dimensional images.

One of the most important human social skills is the ability to recognize faces. Humans recognize familiar faces easily, and can learn to identify unfamiliar faces from repeatedly presented images. Sheep are social animals that can recognize other sheep as well as familiar humans. Little is known, however, about their holistic face-processing abilities. In this study, we trained eight sheep (Ovis aries) to recognize the faces of four celebrities from photographic portraits displayed on computer screens. After training, the sheep chose the 'learned-familiar' faces rather than the unfamiliar faces significantly above chance. We then tested whether the sheep could recognize the four celebrity faces if they were presented in different perspectives. This ability has previously been shown only in humans. Sheep successfully recognized the four celebrity faces from tilted images. Interestingly, there was a drop in performance with the tilted images (from 79.22 ± 7.5% to 66.5 ± 4.1%) of a magnitude similar to that seen when humans perform this task. Finally, we asked whether sheep could recognize a very familiar handler from photographs. Sheep identified the handler in 71.8 ± 2.3% of the trials without pretraining. Together these data show that sheep have advanced face-recognition abilities, comparable with those of humans and non-human primates

Monitoring the dynamics of Src activity in response to anti-invasive dasatinib treatment at a subcellular level using dual intravital imaging

Optimising response to tyrosine kinase inhibitors in cancer remains an extensive field of research. Intravital imaging is an emerging tool, which can be used in drug discovery to facilitate and fine-tune maximum drug response in live tumors. A greater understanding of intratumoural delivery and pharmacodynamics of a drug can be obtained by imaging drug target-specific fluorescence resonance energy transfer (FRET) biosensors in real time. Here, we outline our recent work using a Src-FRET biosensor as a readout of Src activity to gauge optimal tyrosine kinase inhibition in response to dasatinib treatment regimens in vivo. By simultaneously monitoring both the inhibition of Src using FRET imaging, and the modulation of the surrounding extracellular matrix using second harmonic generation (SHG) imaging, we were able to show enhanced drug penetrance and delivery to live pancreatic tumors. We discuss the implications of this dual intravital imaging approach in the context of altered tumor-stromal interactions, while summarising how this approach could be applied to assess other combination strategies or tyrosine kinase inhibitors in a preclinical setting

Heterogeneity in the pancreatic cancer microenvironment – TGFβ as a master regulator?

Pancreatic ductal adenocarcinoma is an aggressive disease for which there are very few available therapies. It is notable for its high degree of tumour complexity, with the tumour microenvironment often accounting for the majority of the tumour volume. Until recently, the biology of the stroma was poorly understood, particularly in terms of heterogeneity. Recent research, however, has shed light on the intricacy of signalling within the stroma and particularly the molecular and functional heterogeneity of the cancer associated fibroblasts. In this review, we summarise the recent improvements in our understanding of the different fibroblast populations within PDAC, with a focus on the role TGFβ plays to dictate their formation and function. These studies have highlighted some of the reasons for the failure of trials targeting the tumour stroma, however, there are still considerable gaps in our knowledge, and more work is needed to make effective fibroblast targeting a reality in the clinic

Combating pancreatic cancer with PI3K pathway inhibitors in the era of personalised medicine

Pancreatic ductal adenocarcinoma (PDAC) is among the most deadly solid tumours. This is due to a generally late-stage diagnosis of a primarily treatment-refractory disease. Several large-scale sequencing and mass spectrometry approaches have identified key drivers of this disease and in doing so highlighted the vast heterogeneity of lower frequency mutations that make clinical trials of targeted agents in unselected patients increasingly futile. There is a clear need for improved biomarkers to guide effective targeted therapies, with biomarker-driven clinical trials for personalised medicine becoming increasingly common in several cancers. Interestingly, many of the aberrant signalling pathways in PDAC rely on downstream signal transduction through the mitogen-activated protein kinase and phosphoinositide 3-kinase (PI3K) pathways, which has led to the development of several approaches to target these key regulators, primarily as combination therapies. The following review discusses the trend of PDAC therapy towards molecular subtyping for biomarker-driven personalised therapies, highlighting the key pathways under investigation and their relationship to the PI3K pathway