Integrated Approach with Genetic Editing and Innovative 3D Models for Podocyte Adhesome Evaluation

• Grant number: 25/09054-3
• Support Opportunities: Research Grants - Initial Project
• Start date: March 01, 2026
• End date: February 28, 2031
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Mara Sanches Guaragna
• Grantee: Mara Sanches Guaragna
• Host Institution: Centro de Biologia Molecular e Engenharia Genética (CBMEG). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• City of the host institution: Campinas
• Associated researchers: Aline Mara dos Santos ; Ana Carolina Migliorini Figueira ; Andre Alexandre de Thomaz ; Ângela Saito ; Luiz Fernando Onuchic ; Marcio Chaim Bajgelman ; Moin Saleem ; Professor Gavin Iain Welsh ; Taís Nitsch Mazzola ; Talita Souza Siqueira
• Associated scholarship(s): 26/07064-4 - Expression and localization of beta-2 syntropin in human podocytes in 2D culture, BP.IC; 26/07094-0 - EXPRESSION OF THE PLS3, TMOD3 AND S100A11 PROTEINS IN HUMAN PODOCYTES IN 2D CULTURE, BP.MS

Abstract

Podocytes are specialized epithelial cells with actin-rich extensions known as foot processes, which are essential for anchoring the cells to the glomerular basement membrane (GBM) within the glomerular filtration barrier (GFB). Retraction, effacement, or detachment of these foot processes from the GBM is a hallmark of glomerular injury and a key indicator of progression toward chronic kidney disease (CKD). CKD is a major global health concern, affecting approximately 10% of the global population, with up to 90% of cases attributed to dysfunction of the GFB. The attachment of foot processes to the GBM is mediated by focal adhesions (FAs), which are components of the integrin adhesome. Disruption of FAs can result in foot process effacement and detachment from the GBM, leading to proteinuria and the progression of CKD - key features of podocytopathies. Despite advances in the field, it remains unclear whether FA components in podocytes are specific to these cells and whether they contribute to the assembly of a unique GBM. In this context, and with the aim of advancing our understanding of podocyte-specific adhesome pathways, this project proposes to study candidate FA proteins using innovative 3D glomerular models. The project will consist of four stages: Stage 1 aims to characterize the expression and localization of five FA protein candidates in 2D podocytes. Proteins in stage 1 will proceed to stage 2, still in 2D podocytes, where they will be: a) knocked out using CRISPR/Cas9 and b) overexpressed using viral vectors. Comparisons between wild-type (WT) and modified (KO and overexpressing) podocytes will be conducted by analyzing cytoskeletal profiles, cell adhesion and motility, and colocalization with binding-partner proteins. Protein(s) which knockout or overexpression result(s) in a significantly altered phenotype will be selected for stage 3, during which two 3D models will be developed: a) glomerular spheroids - Glomspheres, and b) bio-printed biomimetic glomeruli - Glombioprinted for inactivation or overexpression. In stage 4, proteomic analysis will be performed on WT Glomspheres and modified Glomspheres (KO or overexpressing) to compare differentially expressed pathways and identify potential therapeutic targets. Given the complex architecture of glomerular tissue, this ambitious and technically challenging study aims to drive technological innovation by employing advanced 3D cell culture models that more accurately mimic the in vivo glomerular microenvironment, while also advancing our understanding of podocyte-specific adhesome regulation and uncovering novel precision therapeutic targets. (AU)