ADAM12 is a circulating marker for stromal activation in pancreatic cancer and predicts response to chemotherapy.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by abundant stroma that harbors tumor-promoting properties. No good biomarkers exist to monitor the effect of stromal targeting therapies or to predict response. We set out to identify such non-invasive markers for PDAC stroma and predict response to therapy. Gene expression datasets, co-culture experiments, xenografts, and patient samples were analyzed. Serum samples were measured from a cohort of 58 resected patients, and 87 metastatic or locally advanced PDAC patients. Baseline and follow-up levels were assessed in 372 additional metastatic PDAC patients who received nab-paclitaxel with gemcitabine (n = 184) or gemcitabine monotherapy (n = 188) in the phase III MPACT trial. Increased levels of ADAM12 were found in PDAC patients compared to healthy controls (p < 0.0001, n = 157 and n = 38). High levels of ADAM12 significantly associated with poor outcome in resected PDAC (HR 2.07, p = 0.04). In the MPACT trial survival was significantly longer for patients who received nab-paclitaxel and had undetectable ADAM12 levels before treatment (OS 12.3 m vs 7.9 m p = 0.0046). Consistently undetectable or decreased ADAM12 levels during treatment significantly associated with longer survival as well (OS 14.4 m and 11.2 m, respectively vs 8.3, p = 0.0054). We conclude that ADAM12 is a blood-borne proxy for stromal activation, the levels of which have prognostic significance and correlate with treatment benefit.

Defining the molecular pathology of pancreatic body and tail adenocarcinoma.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) remains a dismal disease, with very little improvement in survival over the past 50 years. Recent large-scale genomic studies have improved understanding of the genomic and transcriptomic landscape of the disease, yet very little is known about molecular heterogeneity according to tumour location in the pancreas; body and tail PDACs especially tend to have a significantly worse prognosis. The aim was to investigate the molecular differences between PDAC of the head and those of the body and tail of the pancreas. METHODS: Detailed correlative analysis of clinicopathological variables, including tumour location, genomic and transcriptomic data, was performed using the Australian Pancreatic Cancer Genome Initiative (APGI) cohort, part of the International Cancer Genome Consortium study. RESULTS: Clinicopathological data were available for 518 patients recruited to the APGI, of whom 421 underwent genomic analyses; 179 of these patients underwent whole-genome and 96 RNA sequencing. Patients with tumours of the body and tail had significantly worse survival than those with pancreatic head tumours (12·1 versus 22·0 months; P = 0·001). Location in the body and tail was associated with the squamous subtype of PDAC. Body and tail PDACs enriched for gene programmes involved in tumour invasion and epithelial-to-mesenchymal transition, as well as features of poor antitumour immune response. Whether this is due to a molecular predisposition from the outset, or reflects a later time point on the tumour molecular clock, requires further investigation using well designed prospective studies in pancreatic cancer. CONCLUSION: PDACs of the body and tail demonstrate aggressive tumour biology that may explain worse clinical outcomes.

Transcriptional repression by COUP-TF II is dependent on the C-terminal domain and involves the N-CoR variant, RIP13delta1.

COUP-TF II/ARP-1 is an 'orphan' steroid receptor that inhibits basal transcription, and represses trans-activation by the vitamin D, thyroid hormone and retinoid receptors. The molecular basis of repression by COUP-TF II remains obscure. In this study we utilized the GAL4 hybrid system to demonstrate that COUP-TF II contains sequences within the C-terminal region that encode a dominant transcriptional repressor that inhibits the ability of the potent chimeric transactivator GAL4VP16 to induce transcription. Mammalian two hybrid analysis demonstrated that COUP-TF II did not efficiently interact with either interaction domains I or II from N-CoR and RIP13. However, COUP-TF II efficiently interacts with a region comprised of interaction domains I + II from the corepressor, RIP13delta1. Efficient interaction of the orphan receptor with the corepressor was critically dependent on a large region comprised of the C, D and E domains of COUP-TF II, which correlated with the domain that maximally represses transcription. This investigation suggested that the N-CoR variant, RIP13delta1 interacts with a region of COUP-TF II that functions as a dominant transcriptional repressor.

The effects of cyanide and iodoacetate intoxication and ischaemia on enzyme release from the perfused rat heart.

Isolated rat hearts perfused in the presence of iodoacetate show inhibition of glycolysis and release enzymes into the perfusate. Hearts perfused with cyanide, a mitochondrial inhibitor, show acceleration of glycolysis and no enzyme release. The adenine nucleotide content of the iodoacetate, but not the cyanide-perfused hearts was reduced. These results indicate that the membranes were permeable in the former treatment group. The adenylate energy charge and the ATP content of both the cyanide and iodoacetate treatment groups were similar but, as the extent of enzyme release was quite different, it appears that the energy state of the cell was not the prime factor controlling membrane integrity. Isolated perfused hearts were rendered ischaemic by placing a one-way ball valve in the aortic outflow tract. ATP concentration declined, as did ADP after an initial rise of short duration. AMP concentrations rose as the time of ischaemia increased. At the time at which enzyme release was first determined, the intracellular total adenine nucleotide content began to decline, suggesting that the membrane had become permeable to both small and large molecules. Glycolysis was stimulated by the hypoxia induced in the preparation and then this increase became inhibited. The point at which this inhibition was observed was also the point at which membrane permeability was evident. Taken together, the data from these experiments suggest that the energy derived from the activity of the glycolytic pathway may be important to the heart for maintenance of membrane function, particularly in ischaemia.

Antitumor Activity of Fermented Colostrum and Milk.

Male Swiss mice, implanted with Ehrlich ascites tumor cells, were fed each of the following test materials: fresh bovine colostrum, colostrum cultured with Lactobacillus acidophilus , colostrum cultured with Lactobacillus bulgaricus , colostrum cultured with L. bulgaricus and Streptococcus thermophilus , milk cultured with L. acidophilus and milk cultured with L. bulgaricus . Fresh colostrum had no significant effect when fed ad libitum for 7 consecutive days after tumor implantation. Colostrum fermented with L. acidophilus , L. bulgaricus or yogurt culture significantly (P< 0.05) inhibited tumor cell proliferation as indicated by a 16 to 40% decrease in cell counts and a 13 to 35% decrease in DNA synthesis. Similar effects were noted for whole milk fermented with either L. acidophilus or L. bulgaricus .

Lysosomotropic agents. 4. Carbobenzoxyglycylphenylalanyl, a new protease-sensitive masking group for introduction into cells.

Bioactive primary and secondary amines, when acylated with the Z-Gly-Phe group, are transported into pinocytic cells, such as macrophages, P-815 mastocytoma, SV-40 3T3, and leukemia 1210, much faster than the parent compounds. Amines such as lysosomotropic detergents [R. A. Firestone, J. M. Pisano, and R. J. Bonney, J. Med. Chem., 22, 1130 (1979) and nitrogen mustard, which are deactivated by acylation, are unmasked by enzymic action intracellularly, probably in lysosomes because an acidic pH maximum in activity exists which acts only on the L isomer. The added polarity and molecular weight brought about by acylation prevents the amines' normally facile entry into cells by simple diffusion, restricting it to an active-transport mechanism.

A biochemical study of the cotton pellet granuloma in the rat. Effects of dexamethasone and indomethacin.

The subcutaneous implantation of a cotton pellet into a rat results in the formation of a granuloma at the site of the implant. The early events comprise an accumulation of fluid and protein-aceous material together with an infiltration of neutrophils. The granuloma formed by day 7 is characterized by the formation of a vascularized fibrous capsule containing fibroblasts and infiltrating mononuclear cells which are rich in N-acetyl-beta-D-glucosaminidase (NAG). Granuloma development was quantitated by dry weight measurements, and its cellular content was measured by assaying activity of NAG and total nucleic acid content. Nucleic acid determinations showed that cell infiltration into the granuloma took place at a virtually constant rate over a 7-day period. In contrast, the NAG activity did not change significantly until after day 5 when a large increase in the amount of enzyme extractable from the granuloma was seen. Systemic treatment of the animal with dexamethasone or indomethacin resulted in an inhibition of granuloma weight gain, NAG activity and nucleic acid levels. The data suggest that the two drugs acted during the early phase of granuloma development at the level of cell infiltration. Both drugs given on days 0-3 alone suppressed granuloma formation, whereas treatment on days 4-7 was without effect.

Cultured neonate rat myocytes as a model for the study of myocardial ischaemic necrosis.

The preparation of cultures of neonate rat heart muscle cells is described. These cultures, when subjected to anoxia, show enzyme release that can be directly related to the uptake of a vital dye such trypan blue. Enzyme release is a valid method of estimating cell necrosis in this model. The survival of anoxic cultures is closely associated with glycolytic activity. Glycolysis rate falls and enzyme release increases as the medium glucose concentration is reduced. If glycolysis is inhibited by either 2-deoxyglucose or L-lactate, enzyme release under anoxic conditions is enhanced. Enzyme release correlates inversely with glycolytic activity and the intracellular ATP content of the cultures. Addition of ATP to anoxic cultures partially ameliorates the effect of the anoxia on enzyme release. Elevation of the calcium content of the culture medium exacerbates the damage caused to cardiac myocytes by anoxic insult. This effect can be obtunded by calcium-antagonist drugs such as verapamil or nifedipine and can be explained in terms of reduction in utilization of intracellular ATP by the anoxic monocytes. These observations indicate that cultured myocytes may represent a useful model of hypoxic injury against which novel pharmacological agents, that may reduce hypoxic or ischaemic injury in vivo, could be evaluated.