Base editors as new personalized treatment for Hemophilia A

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Abstract:

Hemophilia A (HA) is an inherited bleeding disorder due to mutations in the F8 gene encoding for factor VIII (FVIII) protein, essential for blood coagulation and hemostasis. People with severe HA experience spontaneous and uncontrolled bleedings that lead to debilitating arthropathies and may be fatal. Patient treatment is based on life-long prophylactic administration of recombinant FVIII products. The quest for new therapeutic options led to the development of new different therapeutic strategies, ranging from recombinant antibodies to gene therapy and genome editing approaches. However, they have not been optimized yet or possess different drawbacks that still need to be addressed. In this context, we propose the exploitation of the recently developed base editing approach as a novel personalized tool to correct the most represented class of pathogenic mutations in HA as well as other inherited monogenic diseases, namely the single nucleotide changes, or point mutations. Base editors (BE), constituted by the fusion of Cas9 nickase with a deaminase enzyme, can virtually target each nucleotide within the genome and, by not inducing any DNA double strand breaks, has very low side effects. Compared to other gene transfer approaches, base editing offers the advantage of correcting pathogenic mutations in large genes, that could not be approached by gene transfer, and of maintaining the physiologic expression of the target gene. Here, we propose to exploit the DNA base editing approach to revert and correct two representative F8 mutations, R593C and R2166*, both associated with severe HA phenotypes. This project aims to achieve a proof-of-concept in already available and ad-hoc generated cellular, ex-vivo and in-vivo mouse models of HA. By directly acting on genomic DNA in a site-specific way, base editing would ensure effective and stable reconstitution of the natural FVIII expression with a single intervention even to young pediatric patients and may result in a life-long cure for people with HA. We believe that the project, through a solid, groundbreaking network with Italian and international actors, will provide with tremendous translational implications, with profound socio-economic impact for healthcare system as well as patient management. We foresee the creation of many opportunities for inventions as well as innovative research tools. The project will be able to attract highly motivated and skilled personnel as well as national and international funds and industrial partners to pre-clinically and clinically develop the proposed base editing approach for Hemophilia, which could be translated further to other inherited disorders. Overall, we aim at providing evidence that the base editing, by fulfilling the longstanding aspiration to make targeted changes in the genome with negligible side effects, has the potential to represent an innovative therapeutic option for HA as well as for the vast majority of monogenic disorders.

Dettagli progetto:

Referente scientifico: Balestra Dario

Fonte di finanziamento: Bando PRIN 2022 

Data di avvio: 05/10/2023

Data di fine: 05/10/2025

Contributo MUR: 96.156 €

Co-finanziamento UniFe: 2.218 €

Partner:

  • Università degli Studi di FERRARA (capofila)
  • Libera Università "Vita Salute S. Raffaele" MILANO