Construction and validation of an RNA trans-splicing molecule suitable to repair a large number of COL7A1 mutations.

RNA trans-splicing has become a versatile tool in the gene therapy of monogenetic diseases. This technique is especially valuable for the correction of mutations in large genes such as COL7A1, which underlie the dystrophic subtype of the skin blistering disease epidermolysis bullosa. Over 800 mutations spanning the entire length of the COL7A1 gene have been associated with defects in type VII collagen, leading to excessive fragility of epithelial tissues, the hallmark of dystrophic epidermolysis bullosa (DEB). In the present study, we designed an RNA trans-splicing molecule (RTM) that is capable of repairing any given mutation within a 4200 nucleotide region spanning the 3' half of COL7A1. The selected RTM, RTM28, was able to induce accurate trans-splicing into endogenous COL7A1 pre-mRNA transcripts in a type VII collagen-deficient DEB patient-derived cell line. Correct trans-splicing was detected at the RNA level by semiquantitative RT-PCR and correction of full-length type VII collagen was confirmed at the protein level by immunofluorescence and western blot analyses. Our results demonstrate that RTM28, which covers >60% of all mutations reported in DEB and is thus the longest RTM described so far for the repair of COL7A1, represents a promising candidate for therapeutic applications.

Assessing vaccine hesitancy and vaccine literacy among the European prison population and staff: A multicentre observational study.

Vaccination is the most efficient and cost-effective public health intervention. Prison population, for its low social distancing, constant turnover, and high percentage of migrants, should be an important target of vaccination campaign. However, vaccination coverage in prison is low. In this study we estimated vaccine hesitancy and vaccine literacy among the prison population and staff and assessed their correlation. We conducted a cross-sectional study in 13 prisons of 4 European countries. The sample included 847 people living in prison and 755 staff members. Through a structured questionnaire we assessed vaccine hesitancy, vaccine literacy, general health literacy, previous vaccine refusal and socio-demographic characteristics of participants. Exploratory factor analysis was used to extract three components of vaccine hesitancy. Logistic regression was applied to assess the association between previous vaccine refusal and vaccine hesitancy; linear regression was applied to assess the association between vaccine hesitancy and vaccine and general health literacy. All analyses were adjusted for socio-demographic variables. We identified three components of vaccine hesitancy explaining 49% of the total variance: Mistrust, Concern and Conspiracy. In both people living in prison and staff, all the components were associated to previous vaccine refusal (p-value < 0.001) and presented good internal consistency (Cronbach's alpha = 0.90, 0.73 and 0.78). Young participants presented the highest levels of vaccine hesitancy; migrant people living in prison had the lower levels of Mistrust and the higher level of Concern; all three factors were lower among participants with the highest degree of education. Mistrust and Concern were inversely associated with vaccine literacy while all three subscales were inversely associated with general health literacy (all p-values < 0.001). This study suggests that educational interventions aimed at increasing vaccine literacy in people living and working in prison could decrease vaccine hesitation and consequently increase vaccination uptake among the prison population and staff.