Valproic acid disables the Nrf2 anti-oxidant response in acute myeloid leukaemia cells enhancing reactive oxygen species-mediated killing.

BACKGROUND: We previously demonstrated the in vitro killing of AML cells by the combination of the lipid-lowering agent bezafibrate (BEZ) and the contraceptive hormone medroxyprogesterone acetate (MPA). A phase II trial demonstrated in vivo safety and efficacy of BEZ and MPA (BaP) in elderly, relapsed/refractory AML and high-risk myelodysplastic syndrome (MDS) patients. However, we observed dose-limiting toxicities in a second trial that attempted to improve outcomes via escalation of BaP doses. Thus we sought to identify a third repurposed drug that potentiates activity of low dose BaP (BaP 0.1 mM). METHODS AND RESULTS: We demonstrate that addition of a commonly used anti-epileptic, valproic acid (VAL) to low dose BaP (BaP 0.1 mM)(VBaP) enhanced killing of AML cell lines/primary AML cells to levels similar to high dose BaP (BaP 0.5 mM). Similarly, addition of VAL to BaP 0.1 mM enhanced reactive oxygen species (ROS), lipid peroxidation and inhibition of de novo fatty acid synthesis. Overexpression of Nrf2 in K562 and KG1a completely inhibited ROS production and rescued cells from VAL/BaP 0.1 mM/VBaP killing. CONCLUSIONS: Given the good safety data of low-dose BaP in elderly/relapsed/refractory AML patients, and that VAL alone is well-tolerated, we propose VBaP as a novel therapeutic combination for AML.

Novel immunotherapeutics against LGR5 to target multiple cancer types.

We have developed and validated a highly specific, versatile antibody to the extracellular domain of human LGR5 (α-LGR5). α-LGR5 detects LGR5 overexpression in >90% of colorectal cancer (CRC), hepatocellular carcinoma (HCC) and pre-B-ALL tumour cells and was used to generate an Antibody-Drug Conjugate (α-LGR5-ADC), Bispecific T-cell Engager (α-LGR5-BiTE) and Chimeric Antigen Receptor (α-LGR5-CAR). α-LGR5-ADC was the most effective modality for targeting LGR5+ cancer cells in vitro and demonstrated potent anti-tumour efficacy in a murine model of human NALM6 pre-B-ALL driving tumour attrition to less than 1% of control treatment. α-LGR5-BiTE treatment was less effective in the pre-B-ALL cancer model yet promoted a twofold reduction in tumour burden. α-LGR5-CAR-T cells also showed specific and potent LGR5+ cancer cell killing in vitro and effective tumour targeting with a fourfold decrease in pre-B-ALL tumour burden relative to controls. Taken together, we show that α-LGR5 can not only be used as a research tool and a biomarker but also provides a versatile building block for a highly effective immune therapeutic portfolio targeting a range of LGR5-expressing cancer cells.

Cell-autonomous Hedgehog signaling controls Th17 polarization and pathogenicity.

Th17 cells are key drivers of autoimmune disease. However, the signaling pathways regulating Th17 polarization are poorly understood. Hedgehog signaling regulates cell fate decisions during embryogenesis and adult tissue patterning. Here we find that cell-autonomous Hedgehog signaling, independent of exogenous ligands, selectively drives the polarization of Th17 cells but not other T helper cell subsets. We show that endogenous Hedgehog ligand, Ihh, signals to activate both canonical and non-canonical Hedgehog pathways through Gli3 and AMPK. We demonstrate that Hedgehog pathway inhibition with either the clinically-approved small molecule inhibitor vismodegib or genetic ablation of Ihh in CD4+ T cells greatly diminishes disease severity in two mouse models of intestinal inflammation. We confirm that Hedgehog pathway expression is upregulated in tissue from human ulcerative colitis patients and correlates with Th17 marker expression. This work implicates Hedgehog signaling in Th17 polarization and intestinal immunopathology and indicates the potential therapeutic use of Hedgehog inhibitors in the treatment of inflammatory bowel disease.

Modality-specific training adaptations - do they lead to a dampened acute inflammatory response to exercise?

While adaptations to a short-term training program can dampen the acute inflammatory response to exercise, less is known about the influence of chronic modality-specific adaptations to training. This study compares the acute inflammatory response to upper- and lower-body interval exercise in individuals chronically trained in these respective modalities. Ninety minutes of interval exercise matched for relative power output on an arm-crank (ARM) and cycle ergometer (LEG) was performed by 8 trained paddlers and 8 trained cyclists. Blood samples were taken before and after exercise. Interleukin-6 (IL-6) concentrations were analysed in plasma, while the expression of intracellular heat shock protein 72 (iHsp72) was assessed in monocytes. IL-6 was increased following both modalities (fold change - ARM: 7.23 ± 3.56, p < 0.001; LEG: 9.03 ± 4.82, p < 0.001), in both groups (cyclists, p < 0.001; paddlers, p < 0.001), but the increase was smaller in ARM compared with LEG (time × modality, p < 0.001). ARM induced a smaller iHsp72 response compared with LEG (fold change - ARM: 1.07 ± 0.14, p = 0.102; LEG: 1.18 ± 0.14, p < 0.001; time × modality, p = 0.039). Following ARM, iHsp72 expression was increased in the cyclists only (fold change cyclists: 1.12 ± 0.11, p = 0.018; paddlers: 1.03 ± 0.17, p = 0.647), while iHsp72 expression following LEG was increased in both groups (fold change cyclists: 1.14 ± 0.15, p = 0.027; paddlers: 1.22 ± 0.13, p < 0.001). Taken together, the acute inflammatory response to lower-body interval exercise was larger compared with work-matched upper-body interval exercise. Moreover, adaptations to upper-body exercise training dampened the iHsp72 response to this modality. Therefore, exercise may be less effective in reducing chronic low-grade inflammation in individuals relying on their upper body, such as wheelchair users.