DEVELOPMENT OF AN IN VIVO MULTITASKING BIOLUMINESCENT REPORTER GENE PLATFORM FOR THE PRECLINICAL EVALUATION OF RNA THERAPEUTICS
type: CreativeWork
description: RNA-based therapeutics have emerged as a transformative class of medicines, enabling precise modulation of gene expression through sequence-defined mechanisms and offering novel therapeutic opportunities for previously intractable diseases. The potential of these drugs to address unmet pharmacological needs, together with their novelty and pharmacokinetic and pharmacodynamic properties that often differ substantially from those of conventional small chemical molecules, makes their preclinical evaluation particularly challenging. Consequently, there is an urgent need for innovative preclinical platforms capable of capturing their complex biological effects beyond conventional toxicity and efficacy endpoints. This PhD thesis is aimed at developing and applying a reporter-based strategy to improve the rational design and preclinical evaluation of RNA-based therapeutics. By integrating in vitro reporter assays with in vivo bioluminescent reporter mouse models, this work enables non-invasive, longitudinal monitoring of key biological pathways involved in pato-physiological mechanisms including inflammation, oxidative stress, cell-cycle regulation, and immunoregulatory signalling. Firstly, in vitro reporter-based screening platform was established to assess off-target effects of the allele-specific siRNA siRyR2-U10, developed for the treatment of catecholaminergic polymorphic ventricular tachycardia (CPVT). Using multiple murine and human cell models, the effects of the siRNA and its calcium phosphate nanoparticle formulation were systematically evaluated across inflammatory, Th2 immune response, oxidative stress, and proliferative pathways. This multidimensional analysis revealed limited immunostimulatory and oxidative stress responses, mainly related to the calcium- phosphate nanoparticles formulation. In contrast, we identified a reproducible, sequence-dependent modulation of cell-cycle progression, thereby laying the groundwork for the use of high-throughput reporter …
abstract: RNA-based therapeutics have emerged as a transformative class of medicines, enabling precise modulation of gene expression through sequence-defined mechanisms and offering novel therapeutic opportunities for previously intractable diseases. The potential of these drugs to address unmet pharmacological needs, together with their novelty and pharmacokinetic and pharmacodynamic properties that often differ substantially from those of conventional small chemical molecules, makes their preclinical evaluation particularly challenging. Consequently, there is an urgent need for innovative preclinical platforms capable of capturing their complex biological effects beyond conventional toxicity and efficacy endpoints. This PhD thesis is aimed at developing and applying a reporter-based strategy to improve the rational design and preclinical evaluation of RNA-based therapeutics. By integrating in vitro reporter assays with in vivo bioluminescent reporter mouse models, this work enables non-invasive, longitudinal monitoring of key biological pathways involved in pato-physiological mechanisms including inflammation, oxidative stress, cell-cycle regulation, and immunoregulatory signalling. Firstly, in vitro reporter-based screening platform was established to assess off-target effects of the allele-specific siRNA siRyR2-U10, developed for the treatment of catecholaminergic polymorphic ventricular tachycardia (CPVT). Using multiple murine and human cell models, the effects of the siRNA and its calcium phosphate nanoparticle formulation were systematically evaluated across inflammatory, Th2 immune response, oxidative stress, and proliferative pathways. This multidimensional analysis revealed limited immunostimulatory and oxidative stress responses, mainly related to the calcium- phosphate nanoparticles formulation. In contrast, we identified a reproducible, sequence-dependent modulation of cell-cycle progression, thereby laying the groundwork for the use of high-throughput reporter …
author: PANZERI, ALESSIA
language: eng
keywords: Settore BIOS-11/A - Farmacologia
date published: 2026-05-14
date modified: 2026-07-27T20:18:56Z
provider: Università degli Studi di Milano
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: Università degli Studi di Milano

PropertyValue
property ID: Handle
value: https://hdl.handle.net/2434/1240381