SMART

The development of next-generation sequencing techniques has favored the identification of new genes involved in the development of renal diseases. As a consequence, the continuous discovery of genetic variants of uncertain significance have raised the problem of finding functional tests to understand their possible pathogenetic role. The aim of this study is to use kidney-free renal progenitors cell cultures as a new in vitro diagnostic tool, supporting clinical diagnosis and classification of kidney diseases on a genetic basis, which may be economical, easy to reproduce and that could be used as a personalized model of illness.

This study follows the introduction, at the Meyer pediatric hospital in Florence, of a new genetic screening test for patients suffering from steroid-resistant nephrotic syndrome ¹. This test, through the use of Next Generation Sequencing, allows the simultaneous analysis of the expression of 46 genes, of which 19 genes known to be responsible for causing the disease and 27 candidate genes described as responsible for the onset of proteinuria in animal models and expressed in the glomerular filtration barrier.

At the moment, the identification of genetic variants with uncertain significance, which require a functional verification, needs the construction of transgenes, transfection experiments and functional studies that involve many months of work. Given these needs, the introduction of a rapid and reproducible functional test in vitro becomes essential to accelerate the diagnosis and improve the clinical approach to the patient.

These cells represent an innovative, personalized tool for the modeling of genetic renal diseases in the context of the complex individual genetic and epigenetic background. Furthermore, patient-specific renal progenitor cells can be used to screen new therapeutic treatment strategies in terms of safety and efficacy. In summary, in this project, we propose the application of a new in vitro diagnostic tool, which will allow a more precise, more reliable, more convenient and early diagnosis of genetic kidney diseases, leading to improved clinical decision making and the prognosis of pediatric patients.

¹. Giglio S et al. Heterogeneous Genetic Alterations in Sporadic Nephrotic Syndrome Associate with Resistance to Immunosuppression. J Am Soc Nephrol. Jul 24, 2014.