Project Details
Projekt
Exploring the Kidney Immune Cell Crosstalk using functionalized kidney organoids, transcriptomics and advanced morphometrics
Subject Area
Pathology
Term
since 2025
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 445703531
Kidney diseases, including acute kidney injury (AKI) and its transition to chronic kidney disease (CKD), are driven by altered immune cell crosstalk. The exact mechanisms of immune cell contribution towards pro-fibrotic disease transition are incompletely understood, in part due to limited availability of (fresh) human tissue. We have demonstrated an intricate cell-crosstalk in the kidney and bone marrow in mice, in which platelets activate and modulate leukocytes to initiate a fibrotic response in stromal cells via CXCL4. We developed a new approach for sequencing physically interacting cells (PIC-seq), which allows deep molecular characterization of cell-cell communication. We used humanized kidney organoids, to study pro-fibrotic disease transitions. We hypothesize that the functional and morphometric analysis of organoids functionalized with an immune compartment and human samples will enable us to better understand (immune) cell interactions in AKI driving the transition to CKD and kidney fibrosis. We will combine our expertise in fibrosis, cellular crosstalk, and kidney organoids (Schneider), with expertise in kidney and autopsy pathology and organ/tissue imaging (von Stillfried). In WP1, we will refine our humanized induced pluripotent stem cell (iPSC)-based kidney organoids with an immune cell compartment (with P1). We will develop different injury models, e.g., using ischemic insults to mimic AKI. iPSC-derived immune cells, specifically mononuclear phagocytes and platelets, will be incorporated under controlled flow conditions to simulate post-injury cellular interactions. In WP2, we will validate the organoid models by comparison with human kidney tissues, including the rapid autopsy cohort (with P3) and the National Autopsy Network (NATON). We will use pathomics for morphological analysis (with P4) and multiplex imaging (with P9) to assess the spatial context of cellular interactions. We will explore a novel approach of nondestructive ex vivo kidney imaging to facilitate validation studies of cellular crosstalk in the kidney tissue analysis by AI-based methods (with P3 and P4), allowing the generation of 3D morphological data. In WP3, we will explore the transcriptional dynamics of CXCL4+ mononuclear phagocytes and platelets with PDGFRb+ fibrosis-driving cells by PIC-seq during the transition from AKI to fibrosis in the validated organoids with P8 and cryopreserved kidney biopsies from patients with P1. In collaboration with P2, computational approaches will reveal receptor-ligand interactions that drive fibrosis in AKI. Using genetic interventions, e.g., via CRISPR/Cas9, we will validate targets in the cellular crosstalk that drives fibrosis. In conclusion, we aim to develop new approaches enabling a better understanding of the immune cell-driven pro-fibrotic crosstalk in the transition of AKI to CKD, which may lead to new therapeutic strategies for CKD prevention and treatment.
DFG Programme
Clinical Research Units
