Cell-Specific Inflammatory Profile Characterization of the Cell Therapy Candidate Rilparencel

Background: Rilparencel is an autologous cell therapy currently in Phase 3 clinical trials for the treatment of patients with CKD and T2D. Results of Phase 2 clinical trials suggest that rilparencel treatment promotes stabilization of eGFR. Derived from a cortical kidney biopsy, rilparencel composition is primarily renal epithelial cells, and previous studies suggest a therapeutic potential via a paracrine mechanism promoting kidney repair. Rilparencel has been previously reported to modulate the NF-κB pathway, and this study explored our understanding of which cell types/states influence cytokine output. Methods: Rilparencel product manufactured from 17 patients enrolled in a Phase 2 investigative clinical trial of T2D with stage 3a-4 CKD (NCT02086574) was submitted for scRNA-seq, and differentially expressed genes (DEGs) were identified. Conditioned-medium (CM) collected during the manufacturing process was analyzed for secreted cytokine levels (Luminex xMAP, Intelliflex). Cell phenotyping and intracellular cytokine levels were evaluated by flow cytometry using the BD FACS Lyric and Cytek Aurora with analysis by FlowJo. Comparison followed with samples from donated kidneys (n=5; National Disease Research Interchange). Pathway analysis was performed with Panther Go Enrichment Analysis. Results: Starting with ex vivo culture from biopsy, inflammatory cytokine output was reduced during manufacturing. Together, gene expression, intracellular and extracellular cytokine expression, along with cell phenotype marker analysis, characterize the effects of cell type/state that influence cytokine output. Pathway analysis suggests involvement of several potential immune modulatory pathways and supports further evaluation of cell state. Conclusion: Specific cell type/state assessments performed during the manufacturing of rilparencel indicates a reduced inflammatory profile and provides enhanced product characterization knowledge. Funding: Commercial Support - ProKidney LLC