Optimization of mRNA molecules for gene-based therapies


Alfredo Nicosia




Department of Molecular Medicine and Medical Biotechnology, Universita’ Federico II



In vivo transfer of genetic material represents the next generation technology platform for medicines targeting the cause of a genetic disease (Gene Therapy: GT) or exploiting the immune system to elicit a prophylactic or therapeutic response against infectious diseases and cancer (Genetic Vaccine: GV).

In the frame of the WP6.3 we aim at developing novel mRNA sequences to: A) improve in vivo translation and/or immune response, and B) modulate the immune response to achieve improved therapeutic index (Figure below)

  1. Different repertoires of 5’ and 3’ natural and synthetic untranslated regions (UTR) will be assembled alone and in different combinations with a reporter gene in synthetic, modified mRNA and formulated in lipo-nanoparticles (mRNA-LNP). High throughput  screening of transfected cells will be performed. Positive hits will be tested for the ability to improve translation and/or immunogenicity in mice.

  2. The objective of this activity is to modulate the immune response induced by mRNA-based GV and GT treatments by increasing it in the former case and by reducing it in the latter one, thereby resulting in a higher treatment efficacy. This will be done by co-administration of the GV or GT mRNA together with the mRNA for an immunomodulating factor that will work as an encoded “user manual” capable of “informing and teaching” the host immune system to enhance the immune response against the encoded antigen of a GV or to ‘immunologically tolerize’ against the product of the GT, should this be a replacement of the endogenous gene or a molecular tool to correct the host DNA (e.g., a molecular scissor such as Cas9). The immunomodulating factors (both enhancers and inhibitors) will be identified by an unbiased approach consisting of a construction and in vivo screening of a library of mRNA-LNP ‘vectorized’ immunomodulators selected in silico by bioinformatic tools. Following identification of mRNA-LNP based immune enhancers and inhibitors, and in-depth analysis of their mechanism of action, we will get insights into the dark matter of the adaptive immunity, and will be able to decipher the molecular circuits responsible for effective gene delivery treatments in healthy individuals, in subjects affected by genetic diseases, in oncologic patients and in patients affected by senescence associated chronic diseases.


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PI: Salvatore Piano