RNA - Genetherapys
Spoke 3 - Neurodegeneration


The scientific plan of Spoke 3 is comprised of five Work Packages (WPs).
WP 3.1 will describe the transcriptional landscapes of small (miRNAs and piwiRNAs) and long non-coding RNAs, of circular RNAs (coding and non-coding) as well as their interactions with proteins in AD, PD, ALS, and PSP while dedicating special attention to the analysis of their epitranscriptomic modifications through the study of different -omics approaches. WP 3.1 aims to establish new platforms for programmable RNA technologies including those sponging miRNA activities, circular RNAs to deliver neuroprotective factors, lncRNAs able to enhance the translation of an mRNA of interest, and aptamers modifying protein-RNA interaction networks. WP 3.1 will also set up brain organoids from patient-derived iPSCs as a tool to validate the biological activity of RNA-based drugs.
WP 3.2 will prioritize candidate therapeutic RNAs and identify promising RNA targets to streamline the acquisition of preclinical data from patient-derived differentiated neurons or brain organoids in vitro and animal models of the neurodegenerative disease of interest. These include modifiers of RNA-protein interaction networks and tools to down-regulate gene expression or modify splicing patterns. WP 3.2 will also tackle neuroinflammation-developing RNAs able to interfere with its role in pathogenesis. WP 3.2 will establish a pipeline to develop mRNA vaccines for proteins prone to aggregation starting with a-synuclein as proof-of-concept.

WP 3.3 will explore the use of RNA molecules to treat neurodegenerative diseases caused by the pathological expansion of nucleotide repeats. While WP 3.3 aims to interfere with pathological RNA translation, it will also focus on identifying and assessing the therapeutic values of modifiers of poly(Qs) toxicity.
WP 3.4 aims to exploit non-coding RNA biology for precision medicine of neurodevelopmental disorders. Given the large heterogeneity of these diseases, WP 3.4 will focus on a specific repertory of gene mutations while searching for common dysregulated drug targets, as well as linear and circular RNA molecules with therapeutic potentials.
WP 3.5 aims to discover and assess the therapeutic use of RNAs involved in stroke and other diseases of the brain.
In summary, the WPs of Spoke 3 aim to collect preclinical data on the most promising RNAs to reach IND filings and therefore lay down the premises for clinical trials.



In 2018, IIT established a new Genomics and RNA Therapy infrastructure at the new Center for Human Technologies (CHT) in Erzelli, Genoa, where Dr. Stefano Gustincich’s laboratory is located. Together with Professor Andrea Cavalli (IIT Affiliate for Spoke 7), the center is a reference point for all RNA therapy activities at IIT. Inaugurated in 2019, CHT was presented in the presence of local authorities and representatives from the Ministry of Health. In 2020, IIT launched the RNA initiative, which includes 20 research groups at IIT relying on CHT as the HUB for their activities. Fourteen of these groups are involved in CN3, distributed among various Spokes 3, 6, 7, and 8.
The flagship laboratory is state-of-the-art, operational, and open to the activities of other entities affiliated with Spoke 3.


Spoke 3 actively collaborates with Spoke 1 in the sharing of knowledge and tools to confront genetic diseases that involve the central nervous systems. Several ongoing collaborations with project leaders from both spokes, including those interested in the optimization of SINEUP RNAs to treat haploinsufficiencies. Spoke 3 relies on Spoke 6 for the development of new technological platforms of programmable RNAs applied to the nervous system for the identification of post-transcriptional modifications of therapeutic value with nanopore sequencing technology, and for imaging techniques in vivo to study RNA drugs dynamics in cells and tissues. Spoke 7 provides knowledge and computational infrastructure to carry out predictions and simulations of RNA secondary structure and interactions with other nucleic acids and proteins. Spoke 3 is tightly collaborating with Spoke 8 for the optimization of delivery technologies to the central nervous system. In particular, Spoke 3 optimizes the expertise of Spoke 8 in the fields of nanoparticle optimization, microvesicles/exosome loading and delivery, technologies to pass the blood-brain barrier, as well as molecules and peptides for homing nanoparticles to specific neuronal cell types. On the way to IND filing, a crucial step is represented by pharmacokinetics and pharmacodynamics analysis as carried out in Spoke 9. Spoke 10 will be instrumental in streamlining GMP production of RNA drug candidates.