Blood and tissue biomarker analysis in dogs with osteosarcoma treated with palliative radiation and intra-tumoral autologous natural killer cell transfer.

We have previously reported radiation-induced sensitization of canine osteosarcoma (OSA) to natural killer (NK) therapy, including results from a first-in-dog clinical trial. Here, we report correlative analyses of blood and tissue specimens for signals of immune activation in trial subjects. Among 10 dogs treated with palliative radiotherapy (RT) and intra-tumoral adoptive NK transfer, we performed ELISA on serum cytokines, flow cytometry for immune phenotype of PBMCs, and PCR on tumor tissue for immune-related gene expression. We then queried The Cancer Genome Atlas (TCGA) to evaluate the association of cytotoxic/immune-related gene expression with human sarcoma survival. Updated survival analysis revealed five 6-month survivors, including one dog who lived 17.9 months. Using feeder line co-culture for NK expansion, we observed maximal activation of dog NK cells on day 17-19 post isolation with near 100% expression of granzyme B and NKp46 and high cytotoxic function in the injected NK product. Among dogs on trial, we observed a trend for higher baseline serum IL-6 to predict worse lung metastasis-free and overall survival (P = 0.08). PCR analysis revealed low absolute gene expression of CD3, CD8, and NKG2D in untreated OSA. Among treated dogs, there was marked heterogeneity in the expression of immune-related genes pre- and post-treatment, but increases in CD3 and CD8 gene expression were higher among dogs that lived > 6 months compared to those who did not. Analysis of the TCGA confirmed significant differences in survival among human sarcoma patients with high and low expression of genes associated with greater immune activation and cytotoxicity (CD3e, CD8a, IFN-γ, perforin, and CD122/IL-2 receptor beta). Updated results from a first-in-dog clinical trial of palliative RT and autologous NK cell immunotherapy for OSA illustrate the translational relevance of companion dogs for novel cancer therapies. Similar to human studies, analyses of immune markers from canine serum, PBMCs, and tumor tissue are feasible and provide insight into potential biomarkers of response and resistance

Seasonality of Holocene hydroclimate in the Eastern Mediterranean reconstructed using the oxygen isotope composition of carbonates and diatoms from Lake Nar, central Turkey

A positive shift in the oxygen isotope composition (δ18O) of lake carbonates in the Eastern Mediterranean from the early to late Holocene is usually interpreted as a change to drier (reduced P/E) conditions. However, it has also been suggested that changes in the seasonality of precipitation could explain these trends. Here, Holocene records of δ18O from both carbonates and diatom silica, from Lake Nar in central Turkey, provide insights into palaeoseasonality. We show how Δδ18Olakewater (the difference between spring and summer reconstructed δ18Olakewater) was minimal in the early Holocene and for most of the last millennium, but was greater at other times. For example, between ~4,100-1,600 years BP we suggest that increased Δδ18Olakewater could have been the result of relatively more spring/summer evaporation, amplified by a decline in lake level. In terms of change in annual mean δ18O, isotope mass balance modelling shows that this can be influenced by changes in seasonal P/E as well as inter-annual P/E, but lake level falls inferred from other proxies confirm there was a mid Holocene transition to drier climatic conditions in central Turkey

Antigenic and genetic characterization of a divergent African virus, Ikoma lyssavirus

In 2009, a novel lyssavirus (subsequently named Ikoma lyssavirus, IKOV) was detected in the brain of an African civet (Civettictis civetta) with clinical rabies in the Serengeti National Park of Tanzania. The degree of nucleotide divergence between the genome of IKOV and those of other lyssaviruses predicted antigenic distinction from, and lack of protection provided by, available rabies vaccines. In addition, the index case was considered likely to be an incidental spillover event, and therefore the true reservoir of IKOV remained to be identified. The advent of sensitive molecular techniques has led to a rapid increase in the discovery of novel viruses. Detecting viral sequence alone, however, only allows for prediction of phenotypic characteristics and not their measurement. In the present study we describe the in vitro and in vivo characterization of IKOV, demonstrating that it is (1) pathogenic by peripheral inoculation in an animal model, (2) antigenically distinct from current rabies vaccine strains and (3) poorly neutralized by sera from humans and animals immunized against rabies. In a laboratory mouse model, no protection was elicited by a licensed rabies vaccine. We also investigated the role of bats as reservoirs of IKOV. We found no evidence for infection among 483 individuals of at least 13 bat species sampled across sites in the Serengeti and Southern Kenya

Engineering of Chinese hamster ovary cell lipid metabolism results in an expanded ER and enhanced recombinant biotherapeutic protein production

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A tale of two lakes: a multi-proxy comparison of Lateglacial and Holocene environmental change in Cappadocia, Turkey

Individual palaeoenvironmental records represent a combination of regional-scale (e.g. climatic) and site-specific local factors. Here we compare multiple climate proxies from two nearby maar lake records, assuming that common signals are due to regional-scale forcing. A new core sequence from Nar Lake in Turkey is dated by varves and U–Th to the last 13.8 ka. Markedly dry periods during the Lateglacial stadial, at 4.3–3.7 and at 3.2–2.6 ka BP, are associated with peaks in Mg/dolomite, positive δ18O, elevated diatom-inferred electrical conductivity, an absence of laminated sediments and low Quercus/chenopod ratios. Wet phases occurred during the early–mid Holocene and 1.5–0.6 ka BP, characterized by negative δ18O, calcite precipitation, high Ca/Sr ratios, a high percentage of planktonic diatoms, laminated sediments and high Quercus/chenopod ratios. Comparison with the record from nearby Eski Acıgöl shows good overall correspondence for many proxies, especially for δ18O. Differences are related to basin infilling and lake ontogeny at Eski Acıgöl, which consequently fails to register climatic changes during the last 2 ka, and to increased flux of lithogenic elements into Nar Lake during the last 2.6 ka, not primarily climatic in origin. In attempting to separate a regional signal from site-specific ‘noise’, two lakes may therefore be better than one

Data for engineering lipid metabolism of Chinese hamster ovary (CHO) cells for enhanced recombinant protein production

The data presented in this article relates to the manuscript entitled ‘Engineering of Chinese hamster ovary cell lipid metabolism results in an expanded ER and enhanced recombinant biotherapeutic protein production’, published in the Journal Metabolic Engineering [1]. In the article here, we present data examining the overexpression of the lipid metabolism modifying genes SCD1 and SREBF1 in CHO cells by densitometry of western blots and by using mass spectrometry to investigate the impact on specific lipid species. We also present immunofluorescence data at the protein level upon SCD1 and SREBF1 overexpression. The growth profile data during batch culture of control CHO cells and CHO cells engineered to overexpress SCD1 and SREBF1 during batch culture are also reported. Finally, we report data on the yields of model secretory recombinant proteins produced from control, SCD1 or SREBF1 engineered cells using a transient expression systems

Exome sequencing in classic hairy cell leukaemia reveals widespread variation in acquired somatic mutations between individual tumours apart from the signature BRAF V(600)E lesion

In classic Hairy cell leukaemia (HCLc), a single case has thus far been interrogated by whole exome sequencing (WES) in a treatment naive patient, in which BRAF V(600)E was identified as an acquired somatic mutation and confirmed as occurring near-universally in this form of disease by conventional PCR-based cohort screens. It left open however the question whether other genome-wide mutations may also commonly occur at high frequency in presentation HCLc disease. To address this, we have carried out WES of 5 such typical HCLc cases, using highly purified splenic tumour cells paired with autologous T cells for germline. Apart from BRAF V(600)E, no other recurrent somatic mutation was identified in these HCLc exomes, thereby excluding additional acquired mutations as also prevalent at a near-universal frequency in this form of the disease. These data then place mutant BRAF at the centre of the neoplastic drive in HCLc. A comparison of our exome data with emerging genetic findings in HCL indicates that additional somatic mutations may however occur recurrently in smaller subsets of disease. As mutant BRAF alone is insufficient to drive malignant transformation in other histological cancers, it suggests that individual tumours utilise largely differing patterns of genetic somatic mutations to coalesce with BRAF V(600)E to drive pathogenesis of malignant HCLc disease