Developing a Pipeline to Employ RNA-Seq as a Complementary Diagnostic Tool in Rare Diseases (ANTHEM-RNA-Seq)
Atypical Hemolytic Uremic Syndrome, Membranoproliferative Glomerulonephritis, Autosomal Dominant Polycystic Kidney
About this trial
This is an interventional diagnostic trial for Atypical Hemolytic Uremic Syndrome focused on measuring Undiagnosed genetic rare diseases, Whole-exome sequencing, RNA-Sequencing, Molecular diagnosis, Skin-derived fibroblasts
Eligibility Criteria
Healthy subjects. Inclusion Criteria: Male and female > 18 years Written informed consent Exclusion Criteria: Inability to understand the potential risk and benefits of the study Legal incapacity Validation cohort. Inclusion criteria: Male and female > 18 years Genetic diseases affecting RNA levels (frameshifts, stop, large deletions, alteration of canonical splicing sites) Written informed consent Exclusion criteria: Underage patients Inability to understand the potential risk and benefits of the study Legal incapacity Discovery cohort. Inclusion criteria: Male and female patients (children and adults with onset in infancy or early adulthood) with rare genetic undiagnosed diseases Patients with no strong candidates based on previous genetic analysis such as WES, but with clinically suspicion of a genetic rare disease Written informed consent Exclusion criteria: Inability to understand the potential risk and benefits of the study Legal incapacity
Sites / Locations
- Centro di Ricerche Cliniche per le Malattie Rare "Aldo e Cele Daccò"
Arms of the Study
Arm 1
Arm 2
Arm 3
Active Comparator
Experimental
Experimental
Healthy subjects
Validation cohort
Discovery cohort
Five healthy donors will be asked to participate in the study to set up the condition for isolation and culture of skin-derived fibroblasts and to establish the RNA-Seq conditions and profile.
To develop a diagnostic pipeline for isolation and sequencing of mRNA from cultured skin fibroblasts, 10 adult patients with known genetic defects affecting RNA levels and/or splicing will be enrolled, as positive controls.
The second group, the discovery cohort, will be composed of 30 undiagnosed symptomatic patients with clinical suspicion of a genetic disease (both children and adults with onset in infancy or early adulthood) referred to the Clinical Research Center for Rare Diseases "Aldo e Cele Daccò", and for which WES analyses did not reveal any causative genetic alteration. To this end, we plan to recruit around 60 patients, their available parents and/or their available informative relatives who will undergo WES, if not previously done. On the basis of literature and our experience we expect that WES will be resolutive in 40-50% of cases (Lunke et al., 2023). Consequently, we hypothesize to identify 30 patients with a negative WES who will enter the discovery RNA-Seq cohort.