Ataluren improves myelopoiesis and neutrophil chemotaxis by restoring ribosome biogenesis and reducing p53 levels in Shwachman-Diamond syndrome cells.

Shwachman-Diamond syndrome (SDS) is characterized by neutropenia, exocrine pancreatic insufficiency and skeletal abnormalities. SDS bone marrow haematopoietic progenitors show increased apoptosis and impairment in granulocytic differentiation. Loss of Shwachman-Bodian-Diamond syndrome (SBDS) expression results in reduced eukaryotic 80S ribosome maturation. Biallelic mutations in the SBDS gene are found in ~90% of SDS patients, ~55% of whom carry the c.183-184TA>CT nonsense mutation. Several translational readthrough-inducing drugs aimed at suppressing nonsense mutations have been developed. One of these, ataluren, has received approval in Europe for the treatment of Duchenne muscular dystrophy. We previously showed that ataluren can restore full-length SBDS protein synthesis in SDS-derived bone marrow cells. Here, we extend our preclinical study to assess the functional restoration of SBDS capabilities in vitro and ex vivo. Ataluren improved 80S ribosome assembly and total protein synthesis in SDS-derived cells, restored myelopoiesis in myeloid progenitors, improved neutrophil chemotaxis in vitro and reduced neutrophil dysplastic markers ex vivo. Ataluren also restored full-length SBDS synthesis in primary osteoblasts, suggesting that its beneficial role may go beyond the myeloid compartment. Altogether, our results strengthened the rationale for a Phase I/II clinical trial of ataluren in SDS patients who harbour the nonsense mutation.

Modelling methacrylated chitosan hydrogel properties through an experimental design approach: from composition to material properties.

Hydrogels of biopolymers are gradually substituting synthetic hydrogels in tissue engineering applications due to their properties. However, biopolymeric hydrogels are difficult to standardize because of the intrinsic variability of the material and the reversibility of physical crosslinking processes. In this work, we synthesized a photocrosslinkable derivative of chitosan (Cs), namely methacrylated chitosan (CsMA), in which the added methacrylic groups allow the formation of hydrogels through radical polymerization triggered by UV exposure. We then performed a systematic study to link the physical properties of the materials to its preparation parameters to standardize its preparation according to specific applications. We studied the properties of CsMA solutions and the derived hydrogels using a statistical method, namely, response surface method, which allowed us to build empirical models describing material properties in terms of several selected processing factors. In particular, we studied the viscosity of CsMA solutions as a function of CsMA concentration, temperature, and shear rate, while hydrogel compression modulus, morphology, degradation and solubilization were investigated as a function of CsMA concentration, photoinitiator concentration and UV exposure. CsMA solutions resulted in shear thinning and were thus suitable for extrusion-based 3D printing. The CsMA hydrogel was found to be highly tunable, with a stiffness in the 12-64 kPa range, and was stable over a long timeframe (up to 60 days). Finally, the possibility to engineer hydrogel stiffness through an empirical model allowed us to hypothesize a number of possible applications based on the mechanical properties of several biological tissues reported in the literature.