Publications

Overall, PHD1 knockout mice display a metabolic phenotype that generally is deemed protective for cardiovascular disease. Future studies should focus on the efficacy, safety, and gender-specific effects of PHD1 inhibition in humans, and unravel the molecular actors responsible for PHD1-driven, likely intestinal, and regulation of cholesterol metabolism.

<b>Rationale:</b> Congenital diaphragmatic hernia (CDH) results in lung hypoplasia. In severe cases, tracheal occlusion (TO) can be offered to promote lung growth. However, the benefit is limited, and novel treatments are required to supplement TO. VEGF (vascular endothelial growth factor) is downregulated in animal models of CDH and could be a therapeutic target, but its role in human CDH is not known. <b>Objectives:</b> To investigate whether VEGF supplementation could be a suitable treatment for CDH-associated lung pathology. <b>Methods:</b> Fetal lungs from patients with CDH were used to determine pulmonary morphology and VEGF expression. A novel human <i>ex vivo</i> model of fetal lung compression recapitulating CDH features was developed and used to determine the effect of exogenous VEGF supplementation. A nanoparticle-based approach for intrapulmonary delivery of VEGF was developed by conjugating it on functionalized nanodiamonds, which was tested in experimental CDH <i>in vivo</i>. <b>Measurements and Main Results:</b> VEGF expression was downregulated in the distal pulmonary epithelium of human CDH fetuses in conjunction with attenuated cell proliferation. The compression model resulted in impaired branching morphogenesis similar to CDH and downregulation of VEGF expression in conjunction with reduced proliferation of terminal bud epithelial progenitors; these could be reversed by exogenous supplementation of VEGF. Prenatal delivery of VEGF with the functionalized nanodiamond VEGF platform in CDH fetal rats resulted in lung growth and pulmonary arterial remodeling that was complementary to that achieved by TO alone with appearances comparable to healthy controls. <b>Conclusions:</b> This innovative approach could have a significant impact on the treatment of CDH.

&lt;p&gt;Supplementary Figure 5 - Effects of VEGFR-1 signaling ablation on PAN02 tumor progression in body-weight matched mice.&lt;/p&gt;

Our results provide evidence that immaturity of plaque vessels is associated with the degradation of angiogenic factors by haemorrhage-conveyed leucocytes and proteases.

Neutrophils are unusual in their reliance on glycolysis to maintain their energy requirements 1 despite the presence of mitochondria and TCA cycle intermediaries 2 .This metabolic adaptation is thought in part to underpin their survival and anti-microbial function in tissues that are typically hypoxic [3][4][5] .Despite their unique metabolism, little is known about the importance of flux between metabolic pathways in determining neutrophil survival responses.Recent work has demonstrated the importance of the HIF/PHD oxygen-sensing pathway in this regard, identifying both HIF-1α and PHD3 as critical regulators of neutrophil survival in hypoxia 6,7 , with the extended survival of neutrophils in hypoxia being dependent upon HIF-1α expression.In parallel, an expanding body of work has addressed the role of HIF-1α in coordinating macrophage functional responses to pro-inflammatory mediators [8][9][10][11] .This work led to the observation that, in macrophages, LPS causes an intracellular increase in succinate levels resulting in HIF-1α stabilization and enhanced IL-1β signaling 11 .Subsequently, the metabolic re-wiring of anti-microbial (M1) and tissue repair (M2) macrophages has been elucidated, with important consequences of TCA cycle activity and integrity for regulation of NO and N-glycosylation signaling respectively 12 .Whether TCA cycle activity and succinate accumulation regulates HIF-1α activity and hypoxic survival in neutrophils is unknown.

Mitotic spindle assembly is mediated by two processes: a centrosomal and a chromosomal pathway. RanGTP regulates the latter process by releasing microtubule-associated proteins from inhibitory complexes. NuSAP, a microtubule- and DNA-binding protein, is a target of RanGTP and promotes the formation of microtubules near chromosomes. However, the contribution of NuSAP to cell proliferation in vivo is unknown. Here, we demonstrate that the expression of NuSAP highly correlates with cell proliferation during embryogenesis and adult life, making it a reliable marker of proliferating cells. Additionally, we show that NuSAP deficiency in mice leads to early embryonic lethality. Spindle assembly in NuSAP-deficient cells is highly inefficient and chromosomes remain dispersed in the mitotic cytoplasm. As a result of sustained spindle checkpoint activity, the cells are unable to progress through mitosis, eventually leading to caspase activation and apoptotic cell death. Together, our findings demonstrate that NuSAP is essential for proliferation of embryonic cells and, simultaneously, they underscore the importance of chromatin-induced spindle assembly.