Mechanistic characterization of a Drosophila model of paraneoplastic nephrotic syndrome

Purpose: Paraneoplastic syndromes occur in cancer patients and originate from dysfunction of organs at a distance from the tumor or its metastasis. In this study, using a Drosophila gut tumor model, we characterized a mechanism of tumor-induced kidney dysfunction. Methods: snRNA-seq analysis of MT samples with and without Yki gut tumors and snRNA-seq analysis of MT samples with kidney principal cell-specific disregulation of PDGF/VEGF signaling Results: we found that Pvf1, a PDGF/VEGF signaling ligand, secreted by gut tumors activates the PvR/JNK/Jra signaling pathway in the principal cells of the kidney, leading to mis-expression of renal genes and paraneoplastic renal syndrome-like phenotypes Conclusions: Our study describes a novel mechanism by which gut tumors perturb the function of the kidney, which might be of clinical relevance for the treatment of paraneoplastic syndromes. 90 pairs of Drosophila MTs were dissociated for each sample and single nuclei were prepared. Ten thousand nuclei per sample were collected by FACS and loaded into a Chromium Controller (10X Genomics, PN-120223) on a Chromium Single Cell B Chip (10X Genomics, PN-120262), and processed to generate single cell gel beads in emulsion (GEM) according to the manufacturer’s protocol (10X Genomics, CG000183). The library was generated using the Chromium Single Cell 3′ Reagent Kits v3.1 (10X Genomics, PN-1000121) and Chromium i7 Multiplex Kit (10X Genomics, PN-120262) according to the manufacturer’s manual. Quality control for the constructed libraries was performed by Agilent Bioanalyzer High Sensitivity DNA kit (Agilent Technologies, 5067-4626). Quantification analysis was performed by Illumina Library Quantification Kit (KAPA Biosystems, KK4824). The library was sequenced on an Illumina NovaSeq system.