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Next generation versatile and effective AAV-mediated large gene delivery

Project description

Advancing gene therapy for the delivery of large genetic cargos

Gene therapy using adeno-associated viruses (AAV) has shown great promise, with several treatments already available and more in development. However, a major limitation remains: AAV vectors can only carry small genetic payloads, making them unsuitable for treating inherited diseases caused by mutations in large genes. Researchers have explored using inteins (small protein elements that can join protein fragments inside cells) to overcome this barrier. While promising, existing intein-based strategies face challenges, including limited efficiency in certain applications, complex design and unwanted by-products. In this context, the ERC-funded NextGeneTx seeks to optimise AAV-intein technology to create a next-generation AAV platform for treating genetic disorders affecting large genes.

Objective

Despite 6 products on the market and dozens in advanced phases of clinical development, in vivo gene therapy with vectors derived from adeno-associated viruses (AAV) still faces the major challenge of the limited vector cargo capacity (< 5kb). This prevents AAV application for treatment of inherited diseases caused by mutations in large genes. To address this, we have recently developed an AAV-based strategy relying on the use of short protein elements called inteins. These, when fused at the extremities of fragments of a large protein delivered to cells through AAV, can mediate joining of the fragments in a traceless manner, resulting in the reconstitution of the intact target protein. However, despite being extensively utilized as biotechnological tools in vitro, the in vivo application of inteins for therapeutic purposes comes with several limitations. These include: insufficient levels of full-length protein reconstitution for some applications; complex design; and production of undesired, potentially harmful by-products. NextGeneTx aims to expand the potential of AAV-inteins as an innovative in vivo biotechnological tool for therapeutic purposes, by addressing each of these limitations. To achieve this, we will leverage cutting-edge technologies to: i. engineer inteins with improved efficiency and greater design flexibility; ii. modulate intracellular trafficking of intein-containing protein fragments to enhance the efficiency and accuracy of full-length protein reconstitution; iii. develop strategies to reduce the production of unwanted by-products. We will test the therapeutic relevance of evolved AAV-inteins in both gene therapy and genome editing applications, using human retinal organoids and animal models of retinal diseases. The outcomes of NextGeneTx will define a highly adaptable, safe, and effective next-generation AAV-based approach to deliver large genes, thus significantly expanding the patient population that can benefit from in vivo gene therapy.

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Call for proposal

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(opens in new window) ERC-2024-STG

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Host institution

FONDAZIONE TELETHON ETS
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 1 400 000,00
Address
VIA VARESE 16/B
00185 ROMA
Italy

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Region
Centro (IT) Lazio Roma
Activity type
Research Organisations
Links
Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 1 500 000,00

Beneficiaries (3)

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