Sc-RNA seq in familiar Gardner syndrome combined left atrial appendage fibroma reveals APC-C-MYC signaling modulates fibrotic subpopulation remodeling

BackgroundGardners syndrome was once considered a subtype of familial adenomatous polyposis (FAP), and their molecular pathological features remain to be clarified. Familiar Gardners syndrome complicated by a rare giant left atrial appendage aneurysm (LAAA) is an unreported novel type of FAP syndrome, and exploring its causative cellular subtypes and molecular pathological features will provide new insights into the precise treatment of the syndrome. MethodsWhole-exome sequencing was performed in the familial Gardners syndrome patients, and single cell sequencing of left atrial dilatation tumor, intestinal polyp and scalp cyst in the proband was performed to explore the cellular and molecular pathological mechanisms. ResultsExon sequencing indicated the presence of a rare germline variant (c. 4666 dup A, p. Thr1556fs, rs587783031) in APC (the adenomatous polyposis coli gene), which caused changes in the APC related Wnt pathway. Sc-RNA seq in LAAA revealed an increased proportion of fibroblasts and immune cells. We re-clustered fibroblasts and identified five distinct sub-populations in LAAA: cancer associcated fibroblasts (CAF), cardiomyocyte like fibroblasts (CLF), endothelial like fibroblasts (ELF), T_cells like fibroblasts (TLF), and nomal like fibroblasts (NLF). Notably, nomal fibroblasts should constitute the main component of normal cardiac fibroblasts in LA tissues, while CAF mainly dominated in LAAA tissues. The trajectory and RNA velocity analysis revealed LAAA fibroblasts made a preferential transition from the immature phase (CLF, ELF, TLF, and NLF) at the apex of the trajectory to mature phase of tumor-like properties (CAF), indicating normal fibroblast was reprogrammed into CAF in LAAA. These results suggested diseased LA tissue with GS patient appears to display tumor-prone properties. Moreover, Sc-RNA seq in intestinal polyp revealed an increased proportion of T_NK cells, and epithelial cells, while plasma cells, mesenchymal cells, and fibroblasts showed a significantly decreased in this patient. GO terms of intestinal polyp-derived fibroblasts suggested that APC/c-MYC signaling modulates fibrotic subpopulation remodeling in intestinal polyp of GS patient. In addition, Sc-RNA seq in scalp cyst revealed an increased proportion in epithelial cells and T cells in this patient. Furthermore, the expression of APC was lowly expressed in endothelial cell, fibroblasts, melanocytes, luminal epithelial, and T cells and the expression of c-MYC was highly expressed in melanocytes, luminal epithelial, and endothelial cell in scalp cyst. Fibroblasts of three tissues was integrated and re-clustered to evaluate the commone menchanisms of fibroblasts remodeling. We identified three fibroblast subpopulations (FC0, FC1 and FC2), the ratio of FC2 was shown a significantly increased in GS patient, and APC-C-MYC signaling might modulates FC2 subpopulation to proliferate fibroblasts in the occurrence of three GS tissues. ConclusionsUsing large-scale single cell RNA sequencing, cellular landscape and aetiology-specific alterations were identified in the left atrial dilatation tumor, intestinal polyp and scalp cyst of the proband. APC-C-MYC signaling modulates fibrotic subpopulation remodeling in LAAA and intestinal polyp while epithelial subpopulation remodeling in scalp cyst, indicating that syndrome subtypes caused by the same gene mutation in the same individual may still lead to different cellular and gene expression signatures and heterogeneity. This new approach provides a wealth of novel insights into the molecular changes that underlie the cellular processes relevant for cardiac biology and pathophysiology and also shed light on strategies for cell type- and stage-specific intervention in cardiac diseases.