A dyad of lymphoblastic lysosomal cysteine proteases degrades the antileukemic drug L-asparaginase

l-Asparaginase is a key therapeutic agent for treatment of childhood acute lymphoblastic leukemia (ALL). There is wide individual variation in pharmacokinetics, and little is known about its metabolism. The mechanisms of therapeutic failure with l-asparaginase remain speculative. Here, we now report that 2 lysosomal cysteine proteases present in lymphoblasts are able to degrade l-asparaginase. Cathepsin B (CTSB), which is produced constitutively by normal and leukemic cells, degraded asparaginase produced by Escherichia coli (ASNase) and Erwinia chrysanthemi. Asparaginyl endopeptidase (AEP), which is overexpressed predominantly in high-risk subsets of ALL, specifically degraded ASNase. AEP thereby destroys ASNase activity and may also potentiate antigen processing, leading to allergic reactions. Using AEP-mediated cleavage sequences, we modeled the effects of the protease on ASNase and created a number of recombinant ASNase products. The N24 residue on the flexible active loop was identified as the primary AEP cleavage site. Sole modification at this site rendered ASNase resistant to AEP cleavage and suggested a key role for the flexible active loop in determining ASNase activity. We therefore propose what we believe to be a novel mechanism of drug resistance to ASNase. Our results may help to identify alternative therapeutic strategies with the potential of further improving outcome in childhood ALL

Desert dune system response to Late Quaternary environmental change in the northeastern Rub’ al Khali: Advances in the application of optically stimulated luminescence datasets

The application of optically stimulated luminescence (OSL) dating to desert sand dunes has allowed accumulation histories to be used as tools to infer past environmental change. In response to issues facing the interpretation of these records, two research questions are addressed in this thesis. (i) Are dune chronologies representative of dune stratigraphies? And (ii) how can we most appropriately interpret dune chronologies as records of Quaternary environmental conditions? Five dune profiles were sampled for OSL dating at two sites in the northeastern Rub’ al Khali in the southern Arabian Peninsula. The visible stratigraphy was used to guide sampling for three of the profiles and the effectiveness of this approach is assessed. A key finding is that bounding surfaces are not always identifiable as chronological hiatuses by OSL dating, given the level of precision that can be achieved. Using hierarchical relationships visible in two-dimensional exposures is therefore not guaranteed to identify the depositional units necessary to reconstruct dune histories. Comparison of the depositional records from three sampled profiles shows that there is significant variability in chronologies at both the dune and dunefield scales. In light of these findings, the use of ‘range-finder’ OSL dating was investigated as a method of increasing sample throughput in the laboratory. It is concluded that the use of partially prepared samples and shortened measurement techniques can be used to rapidly assess the chronological context of samples and target those units most useful in constructing dune profiles. A new method of presenting dunefield OSL datasets as net accumulation rates, incorporating accumulation thickness rather than relying on the frequency of ages, is presented. Within the last 30 ka, regional accumulation and preservation occurred at ~30-26, 22.5-18, 16-9, 6-2.7, 2.1-1.6, 1.1 and 0.7 ka. In conjunction with numerical model results and a review of other palaeoenvironmental archives, the regional aeolian record is interpreted as a response to changing forcing factors. High rates of net accumulation between ~16-9 ka are attributed to coeval increases in sediment supply and transport capacity. A hiatus in accumulation between ~9-6 ka is interpreted as a result of reduced sediment availability due to high moisture levels. The importance of both external forcing factors and local controls on dune accumulation processes is recognised, and therefore the importance of sampling at multiple locations to distinguish these factors is emphasised.</p

Striatal networks for tinnitus treatment targeting.

Neuromodulation treatment effect size for bothersome tinnitus may be larger and more predictable by adopting a target selection approach guided by personalized striatal networks or functional connectivity maps. Several corticostriatal mechanisms are likely to play a role in tinnitus, including the dorsal/ventral striatum and the putamen. We examined whether significant tinnitus treatment response by deep brain stimulation (DBS) of the caudate nucleus may be related to striatal network increased functional connectivity with tinnitus networks that involve the auditory cortex or ventral cerebellum. The first study was a cross-sectional 2-by-2 factorial design (tinnitus, no tinnitus; hearing loss, normal hearing, n&nbsp;=&nbsp;68) to define cohort level abnormal functional connectivity maps using high-field 7.0&nbsp;T resting-state fMRI. The second study was a pilot case-control series (n&nbsp;=&nbsp;2) to examine whether tinnitus modulation response to caudate tail subdivision stimulation would be contingent on individual level striatal connectivity map relationships with tinnitus networks. Resting-state fMRI identified five caudate subdivisions with abnormal cohort level functional connectivity maps. Of those, two connectivity maps exhibited increased connectivity with tinnitus networks-dorsal caudate head with Heschl's gyrus and caudate tail with the ventral cerebellum. DBS of the caudate tail in the case-series responder resulted in dramatic reductions in tinnitus severity and loudness, in contrast to the nonresponder who showed no tinnitus modulation. The individual level connectivity map of the responder was in alignment with the cohort expectation connectivity map, where the caudate tail exhibited increased connectivity with tinnitus networks, whereas the nonresponder individual level connectivity map did not

Global resting-state functional connectivity of neural oscillations in tinnitus with and without hearing loss

This study examined global resting-state functional connectivity of neural oscillations in individuals with chronic tinnitus and normal and impaired hearing. We tested the hypothesis that distinct neural oscillatory networks are engaged in tinnitus with and without hearing loss. In both tinnitus groups, with and without hearing loss, we identified multiple frequency band-dependent regions of increased and decreased global functional connectivity. We also found that the auditory domain of tinnitus severity, assayed by the Tinnitus Functional Index, was associated with global functional connectivity in both auditory and nonauditory regions. These findings provide candidate biomarkers to target and monitor treatments for tinnitus with and without hearing loss

Effect of mitoxantrone on outcome of children with first relapse of acute lymphoblastic leukaemia (ALL R3): an open-label randomised trial

Background Although survival of children with acute lymphoblastic leukaemia has improved greatly in the past two decades, the outcome of those who relapse has remained static. We investigated the outcome of children with acute lymphoblastic leukaemia who relapsed on present therapeutic regimens. Methods This open-label randomised trial was undertaken in 22 centres in the UK and Ireland and nine in Australia and New Zealand. Patients aged 1—18 years with first relapse of acute lymphoblastic leukaemia were stratified into high-risk, intermediate-risk, and standard-risk groups on the basis of duration of first complete remission, site of relapse, and immunophenotype. All patients were allocated to receive either idarubicin or mitoxantrone in induction by stratified concealed randomisation. Neither patients nor those giving interventions were masked. After three blocks of therapy, all high-risk group patients and those from the intermediate group with postinduction high minimal residual disease (≥10−4 cells) received an allogenic stem-cell transplant. Standard-risk and intermediate-risk patients with postinduction low minimal residual disease (<10−4 cells) continued chemotherapy. The primary outcome was progression-free survival and the method of analysis was intention-to-treat. Randomisation was stopped in December, 2007 because of differences in progression-free and overall survival between the two groups. This trial is registered, reference number ISCRTN45724312. Findings Of 239 registered patients, 216 were randomly assigned to either idarubicin (109 analysed) or mitoxantrone (103 analysed). Estimated 3-year progression-free survival was 35·9% (95% CI 25·9—45·9) in the idarubicin group versus 64·6% (54·2—73·2) in the mitoxantrone group (p=0·0004), and 3-year overall survival was 45·2% (34·5—55·3) versus 69·0% (58·5—77·3; p=0·004). Differences in progression-free survival between groups were mainly related to a decrease in disease events (progression, second relapse, disease-related deaths; HR 0·56, 0·34—0·92, p=0·007) rather than an increase in adverse treatment effects (treatment death, second malignancy; HR 0·52, 0·24—1·11, p=0·11). Interpretation As compared with idarubicin, mitoxantrone conferred a significant benefit in progression-free and overall survival in children with relapsed acute lymphobastic leukaemia, a potentially useful clinical finding that warrants further investigation. Funding Cancer Research UK, Leukaemia and Lymphoma Research, Cancer Council NSW, and Sporting Chance Cancer Foundation.Catriona Parker, Rachel Waters, Carly Leighton, Jeremy Hancock, Rosemary Sutton, Anthony V Moorman, Philip Ancliff, Mary Morgan, Ashish Masurekar, Nicholas Goulden, Nina Green, Tamas Révész, Philip Darbyshire, Sharon Love and Vaskar Sah