Tumour-infiltrating regulatory T cell density before neoadjuvant chemoradiotherapy for rectal cancer does not predict treatment response

Neoadjuvant (preoperative) chemoradiotherapy (CRT) decreases the risk of rectal cancer recurrence and reduces tumour volume prior to surgery. However, response to CRT varies considerably between individuals and factors associated with response are poorly understood. Foxp3+ regulatory T cells (Tregs) inhibit anti-tumour immunity and may limit any response to chemotherapy and radiotherapy. We have previously reported that a low density of Tregs in the tumour stroma following neoadjuvant CRT for rectal cancer is associated with improved tumour regression. Here we have examined the association between Treg density in pre-treatment diagnostic biopsy specimens and treatment response, in this same patient cohort. We aimed to determine whether pre-treatment tumour-infiltrating Treg density predicts subsequent response to neoadjuvant CRT. Foxp3+, CD8+ and CD3+ cell densities in biopsy samples from 106 patients were assessed by standard immunohistochemistry (IHC) and evaluated for their association with tumour regression grade and survival. We found no association between the density of any T cell subset pre-treatment and clinical outcome, indicating that tumour-infiltrating Treg density does not predict response to neoadjuvant CRT in rectal cancer. Taken together with the findings of the previous study, these data suggest that in the context of neoadjuvant CRT for rectal cancer, the impact of chemotherapy and/or radiotherapy on anti-tumour immunity may be more important than the state of the pre-existing local immune response

Local and exotic sources of sarsen debitage at Stonehenge revealed by geochemical provenancing

The application of novel geochemical provenancing techniques has changed our understanding of the construction of Stonehenge, by identifying West Woods on the Marlborough Downs as the likely source area for the majority of the extant sarsen megaliths at the monument. In this study, we apply the same techniques to saccharoid sarsen fragments from three excavations within and outwith the main Sarsen Circle to expand our understanding of the provenance of sarsen debitage present at the monument. Through pXRF analysis, we demonstrate that the surface geochemistry of 1,028 excavated sarsen fragments is significantly affected by subsurface weathering following burial in a way that cannot be overcome by simple cleaning. However, we show that this effect is surficial and does not have a volumetrically significant impact, thus permitting the subsequent use of whole-rock analytical methods. Comparison of ICP-AES and ICP-MS trace element data from 54 representative sarsen fragments with equivalent data from Stone 58 at Stonehenge demonstrates that none are debitage produced during the dressing of this megalith or its 49 chemical equivalents at the monument. Further inspection of the ICP-MS data reveals that 22 of these fragments fall into three distinct geochemical ‘families’. None of these families overlap with the geochemical signature of Stone 58 and its chemical equivalents, implying that sarsen imported from at least a further three locations (in addition to West Woods) is present at Stonehenge. Comparison of immobile trace element signatures from the 54 excavated sarsen fragments against equivalent data for 20 sarsen outcrop areas across southern Britain shows that 15 of the fragments can be linked to specific localities. Eleven of these were likely sourced from Monkton Down, Totterdown Wood and West Woods on the Marlborough Downs (25–33 km north of Stonehenge). Three fragments likely came from Bramdean, Hampshire (51 km southeast of Stonehenge), and one from Stoney Wish, East Sussex (123 km to the southeast). Technological analysis and refitting shows that one of the fragments sourced from Monkton Down was part of a 25.7 cm × 17.9 cm flake removed from the outer surface of a large sarsen boulder, most probably during on-site dressing. This adds a second likely source area for the sarsen megaliths at Stonehenge in addition to West Woods. At this stage, we can only speculate on why sarsen from such diverse sources is present at Stonehenge. We do not know whether the fragments analysed by ICP-MS were removed from (i) the outer surface of Stones 26 or 160 (which are chemically distinct to the other extant sarsen megaliths), (ii) one of the c.28 sarsen megaliths and lintels from the c.60 erected during Stage 2 of the construction of Stonehenge that may now be missing from the monument, or (iii) one of the dismantled and destroyed sarsen megaliths associated with Stage 1 of the monument. With the exception of the fragment sourced from Monkton Down, it is also possible that the analysed fragments were (iv) pieces of saccharoid sarsen hammerstones or their pre-forms, or (v) small blocks brought on-site for ceremonial or non-ceremonial purposes

Distinct molecular signatures of clinical clusters in people with type 2 diabetes:an IMI-RHAPSODY study

Type 2 diabetes is a multifactorial disease with multiple underlying aetiologies. To address this heterogeneity a previous study clustered people with diabetes into five diabetes subtypes. The aim of the current study is to investigate the aetiology of these clusters by comparing their molecular signatures. In three independent cohorts, in total 15,940 individuals were clustered based on five clinical characteristics. In a subset, genetic- (N=12828), metabolomic- (N=2945), lipidomic- (N=2593) and proteomic (N=1170) data were obtained in plasma. In each datatype each cluster was compared with the other four clusters as the reference. The insulin resistant cluster showed the most distinct molecular signature, with higher BCAAs, DAG and TAG levels and aberrant protein levels in plasma enriched for proteins in the intracellular PI3K/Akt pathway. The obese cluster showed higher cytokines. A subset of the mild diabetes cluster with high HDL showed the most beneficial molecular profile with opposite effects to those seen in the insulin resistant cluster. This study showed that clustering people with type 2 diabetes can identify underlying molecular mechanisms related to pancreatic islets, liver, and adipose tissue metabolism. This provides novel biological insights into the diverse aetiological processes that would not be evident when type 2 diabetes is viewed as a homogeneous diseas

ADCY5 couples glucose to insulin secretion in human islets

Single nucleotide polymorphisms (SNPs) within the ADCY5 gene, encoding adenylate cyclase 5, are associated with elevated fasting glucose and increased type 2 diabetes (T2D) risk. Despite this, the mechanisms underlying the effects of these polymorphic variants at the level of pancreatic β-cells remain unclear. Here, we show firstly that ADCY5 mRNA expression in islets is lowered by the possession of risk alleles at rs11708067. Next, we demonstrate that ADCY5 is indispensable for coupling glucose, but not GLP-1, to insulin secretion in human islets. Assessed by in situ imaging of recombinant probes, ADCY5 silencing impaired glucose-induced cAMP increases and blocked glucose metabolism toward ATP at concentrations of the sugar >8 mmol/L. However, calcium transient generation and functional connectivity between individual human β-cells were sharply inhibited at all glucose concentrations tested, implying additional, metabolism-independent roles for ADCY5. In contrast, calcium rises were unaffected in ADCY5-depleted islets exposed to GLP-1. Alterations in β-cell ADCY5 expression and impaired glucose signaling thus provide a likely route through which ADCY5 gene polymorphisms influence fasting glucose levels and T2D risk, while exerting more minor effects on incretin action

Chronic Activation of γ2 AMPK Induces Obesity and Reduces β Cell Function.

Despite significant advances in our understanding of the biology determining systemic energy homeostasis, the treatment of obesity remains a medical challenge. Activation of AMP-activated protein kinase (AMPK) has been proposed as an attractive strategy for the treatment of obesity and its complications. AMPK is a conserved, ubiquitously expressed, heterotrimeric serine/threonine kinase whose short-term activation has multiple beneficial metabolic effects. Whether these translate into long-term benefits for obesity and its complications is unknown. Here, we observe that mice with chronic AMPK activation, resulting from mutation of the AMPK γ2 subunit, exhibit ghrelin signaling-dependent hyperphagia, obesity, and impaired pancreatic islet insulin secretion. Humans bearing the homologous mutation manifest a congruent phenotype. Our studies highlight that long-term AMPK activation throughout all tissues can have adverse metabolic consequences, with implications for pharmacological strategies seeking to chronically activate AMPK systemically to treat metabolic disease

Abstracts from the 8th International Conference on cGMP Generators, Effectors and Therapeutic Implications

This work was supported by a restricted research grant of Bayer AG