title: Human Urine-Derived Renal Progenitors for Personalized Modeling of Genetic Kidney Disorders.
abstract: The critical role of genetic and epigenetic factors in the pathogenesis of kidney disorders is gradually becoming clear, and the need for disease models that recapitulate human kidney disorders in a personalized manner is paramount. In this study, we describe a method to select and amplify renal progenitor cultures from the urine of patients with kidney disorders. Urine-derived human renal progenitors exhibited phenotype and functional properties identical to those purified from kidney tissue, including the capacity to differentiate into tubular cells and podocytes, as demonstrated by confocal microscopy, Western blot analysis of podocyte-specific proteins, and scanning electron microscopy. Lineage tracing studies performed with conditional transgenic mice, in which podocytes are irreversibly tagged upon tamoxifen treatment (NPHS2.iCreER;mT/mG), that were subjected to doxorubicin nephropathy demonstrated that renal progenitors are the only urinary cell population that can be amplified in long-term culture. To validate the use of these cells for personalized modeling of kidney disorders, renal progenitors were obtained from (1) the urine of children with nephrotic syndrome and carrying potentially pathogenic mutations in genes encoding for podocyte proteins and (2) the urine of children without genetic alterations, as validated by next-generation sequencing. Renal progenitors obtained from patients carrying pathogenic mutations generated podocytes that exhibited an abnormal cytoskeleton structure and functional abnormalities compared with those obtained from patients with proteinuria but without genetic mutations. The results of this study demonstrate that urine-derived patient-specific renal progenitor cultures may be an innovative research tool for modeling of genetic kidney disorders.
authors.affiliation: Excellence Centre for Research, Transfer and High Education for the Development of DE NOVO Therapies (DENOTHE) and paola.romagnani@unifi.it elena.lazzeri@unifi.it.
authors.forename: Elena
authors.initials: E
authors.lastname: Lazzeri
authors.affiliation: Excellence Centre for Research, Transfer and High Education for the Development of DE NOVO Therapies (DENOTHE) and Department of Clinical and Experimental Biomedical Sciences, University of Florence, Florence, Italy;
authors.forename: Elisa
authors.initials: E
authors.lastname: Ronconi
authors.affiliation: Excellence Centre for Research, Transfer and High Education for the Development of DE NOVO Therapies (DENOTHE) and.
authors.forename: Maria Lucia
authors.initials: ML
authors.lastname: Angelotti
authors.affiliation: Excellence Centre for Research, Transfer and High Education for the Development of DE NOVO Therapies (DENOTHE) and Department of Clinical and Experimental Biomedical Sciences, University of Florence, Florence, Italy;
authors.forename: Anna
authors.initials: A
authors.lastname: Peired
authors.affiliation: Medical Genetics Unit and.
authors.forename: Benedetta
authors.initials: B
authors.lastname: Mazzinghi
authors.affiliation: Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy;
authors.forename: Francesca
authors.initials: F
authors.lastname: Becherucci
authors.affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri," Centro Anna Maria Astori, Bergamo, Italy; and.
authors.forename: Sara
authors.initials: S
authors.lastname: Conti
authors.affiliation: Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy;
authors.forename: Giulia
authors.initials: G
authors.lastname: Sansavini
authors.affiliation: Excellence Centre for Research, Transfer and High Education for the Development of DE NOVO Therapies (DENOTHE) and.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Sisti
authors.affiliation: Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy;
authors.forename: Fiammetta
authors.initials: F
authors.lastname: Ravaglia
authors.affiliation: Excellence Centre for Research, Transfer and High Education for the Development of DE NOVO Therapies (DENOTHE) and.
authors.forename: Duccio
authors.initials: D
authors.lastname: Lombardi
authors.affiliation: Medical Genetics Unit and.
authors.forename: Aldesia
authors.initials: A
authors.lastname: Provenzano
authors.affiliation: Department of Nephrology, Bellvitge's University Hospital, Barcelona, Spain.
authors.forename: Anna
authors.initials: A
authors.lastname: Manonelles
authors.affiliation: Department of Nephrology, Bellvitge's University Hospital, Barcelona, Spain.
authors.forename: Josep M
authors.initials: JM
authors.lastname: Cruzado
authors.affiliation: Department of Clinical and Experimental Biomedical Sciences, University of Florence, Florence, Italy; Medical Genetics Unit and.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Giglio
authors.affiliation: Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy;
authors.forename: Rosa Maria
authors.initials: RM
authors.lastname: Roperto
authors.affiliation: Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy;
authors.forename: Marco
authors.initials: M
authors.lastname: Materassi
authors.affiliation: Excellence Centre for Research, Transfer and High Education for the Development of DE NOVO Therapies (DENOTHE) and.
authors.forename: Laura
authors.initials: L
authors.lastname: Lasagni
authors.affiliation: Excellence Centre for Research, Transfer and High Education for the Development of DE NOVO Therapies (DENOTHE) and Department of Clinical and Experimental Biomedical Sciences, University of Florence, Florence, Italy; Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy; paola.romagnani@unifi.it elena.lazzeri@unifi.it.
authors.forename: Paola
authors.initials: P
authors.lastname: Romagnani
mesh_terms: D000293:Adolescent; D000818:Animals; D016022:Case-Control Studies; D018929:Cell Culture Techniques; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D007668:Kidney / Q000166:cytology*; D007674:Kidney Diseases / Q000151:congenital*; D008297:Male; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D008822:Mice, Transgenic; D013234:Stem Cells / Q000166:cytology*; D014556:Urine / Q000166:cytology*
keywords: glomerulonephritis; glomerulosclerosis; kidney; mutation; next generation; podocytes