WIP 61 - Differentiation and Proliferation of Neonatal Pulmonary Mesenchymal Cells and Congenital Peribronchial Myofibroblastic Tumor cells with EGFR KDD mutation

Publication Number: WIP 61.7619

Desiree Gutmann, Lucile Packard Children's Hospital Stanford, Palo Alto, CA, United States; Lindsay D.. Butcher, Stanford University School of Medicine, Palo Alto, CA, United States; Dhwani Mehta, Stanford University School of Medicine, Palo Alto, CA, United States; Raya Saab, Stanford University School of Medicine, Palo Alto, CA, United States; Min Huang, MH, Stanford, CA, United States; Serena Y. Tan, Stanford University School of Medicine, Stanford, CA, United States; Lawrence S.. Prince, Stanford University, Palo Alto, CA, United States

Background: Congenital peribronchial myofibroblastic tumor (CPMT) is a rare lung malformation associated with fetal hydrops and acute respiratory failure at birth. CPMTs are thought to arise from mesenchymal progenitor cells that differentiate into cartilage, smooth muscle, or fibroblastic lineages. We are studying patient-derived CPMT cells carrying a novel mutation in the EGF receptor (EGFR). How this mutation alters mesenchymal cell lineage determination and proliferation has not been studied.

Objective: Histological analysis of the resected CPMT identified abnormal cartilage, smooth muscle, myofibroblasts, and fibroblasts. Using CPMT and control primary cells, we are testing how the EGFR KDD mutation alters mesenchymal cell differentiation. Spatial transcriptomics of tumor sections will explore the correlation between cellular differentiation of CPMT cells and EGFR ligand expression within the developing lung niche. These findings will offer insights into altered human lung mesenchymal differentiation.

Design/Methods: We have isolated cells from the CPMT and unaffected adjacent lung. We are currently differentiating CPMT and control cell lines into cartilage, smooth muscle, bone, adipose, and endothelium using defined cell culture media, conditions, and supplements. Cell proliferation and differentiation will be measured by lineage-specific stains and transcriptional profiles. We will next test the impact of EGFR ligands and inhibitors. Once methods are established with control cells, we will begin utilizing patient cells in January 2025. In February/March 2025, we will perform MERFISH to identify transcriptional signatures of tumor cells and adjacent unaffected lung cells that correlate with tumorgenesis, abnormal differentiation, proliferation, and EGFR signaling. No IRB approval is necessary as this is a cell-based study without patient-identifying information. Statistical analysis will compare groups by t-test and ANOVA testing.