Nano-ink based Antisense oligonUcleoTIde deLivery for Ultra-personaIized treatment of Syndromic craniosynostoses (NAUTILUS)

Syndromic craniosynostoses (SCS) are a group of rare genetic disorders characterized by the premature fusion of cranial sutures, leading to severe craniofacial anomalies and restricted brain growth. SCS  include autosomal dominant conditions, such as Muenke, Saethre-Chotzen, Crouzon, Apert, Pfeiffer and craniofrontonasal syndromes, caused by gain-of-function (GoF) or loss-of-function (LoF) mutations in genes critical for cranial suture biology, such as FGFR1/2/3, TWIST1 or TCF12. Current treatments rely exclusively  on invasive surgical interventions, such as early cranial vault remodelling and subsequent maxillofacial procedures, which are associated with significant complications, including severe blood loss, infections, and high rates of resynostosis. Despite technological advancements, currently there are no approved pharmacological adjuvant options for SCS patients, including strategies to address the genetic causes of SCS that drive pathological suture ossification. Recently, RNA interference-based approaches targeting specific SCS-related mutations have shown promising results in restoring the overactive osteogenic differentiation of suture-derived cells. Nevertheless, challenges related to delivery efficiency and to stable and sustained action remain major barriers to clinical translation. NAUTILUS aims to overcome these issues by developing a combined nano-ink platform delivering SCS mutation-specific Antisense Oligonucleotides (ASOs) as a noninvasive, ultra-personalized therapeutic strategy for SCS. To this aim, NAUTILUS will focus on designing and validating patient-specific ASOs tailored to the molecular drivers of SCS. These molecules will be tested for their ability to restore proper target transcript levels and effective protein function in patient-derived cells, thus identifying ASOs able to modulate gene expression, either by silencing pathogenic GoF variants or restoring normal expression in LoF cases. Simultaneously, the project will develop and optimize the nanoink delivery system to ensure controlled and effective release of ASOs over time. This platform combines the recently formulated PLGA-PEG-bis-sulfone nanoparticles for ASOs delivery with a GelMA-based hydrogel scaffold, enabling localized and sustained delivery and release of ASOs directly within the cranial suture microenvironment. Preclinical validation will then be carried out in relevant mouse models of SCS, evaluating the therapeutic platform's ability to delay or prevent suture ossification and mitigate the need for repeated surgical interventions. By targeting the genetic roots of SCS, NAUTILUS aims to revolutionize the clinical management of these conditions. This approach offers the potential to significantly reduce the invasiveness of current treatments, improve surgical outcomes, and enhance the quality of life for affected individuals. Ultimately, the project represents a paradigm shift in the treatment of syndromic craniosynostoses, paving the way for precision medicine in rare craniofacial disorders.