PLCB4 copy gain and PLCβ4 overexpression in primary gastrointestinal stromal tumors: Integrative characterization of a lipid-catabolizing enzyme associated with worse diseasefree survival

Abstract

To explore the implications of lipid catabolism-associated genes in gastrointestinal stromal tumors, we reappraised transcriptomic and genomic datasets and identified copygained and differentially upregulated PLCB4 gene associated with tumor progression. On full sections, PLCB4 mRNA abundance and PLCβ4 immunoexpression were validated in 70 cases. On tissue microarrays, PLCB4 gene copies and PLCβ4 immunoexpression were both informative in 350 cases with KIT/PDGFRA/BRAF genotypes noted in 213. In GIST48 cell line, we stably silenced PLCB4 and YAP1 to characterize their functional effects and regulatory link. Compared with normal tissue, PLCB4 mRNA abundance significantly increased across tumors of various risk levels (p < 0.001), and was strongly correlated with immunoexpression level (p < 0.001, r=0.468). Including polysomy (12.6%) and amplification (17.4%), PLCB4 copy gain was detected in 105 (30%) cases and significantly more frequent (p < 0.001) in cases exhibiting higher PLCβ4 immunoexpression (82/175). Copy gain and protein overexpression were modestly associated with unfavorable genotypes (both p < 0.05), strongly associated with increased size, mitosis, and risk levels defined by both the NIH and NCCN schemes (all p < 0.001), and univariately predictive of shorter disease-free survival (both p < 0.0001). In PLCβ4-overexpressing cases, PLCB4 copy gain still predicted worse prognosis (p < 0.0001). In a multivariate comparison, both overexpression (p=0.007, hazard ratio: 2.454) and copy gain (p=0.031, hazard ratio: 1.892) exhibited independent impact. In vitro, YAP1 increased PLCB4 mRNA and protein expression, and both molecules significantly promoted cell proliferation. Being driven by copy gain or YAP1, PLCβ4 is a novel overexpressed enzyme regulating lipid catabolism that promotes cell proliferation and independently confers a worse prognosis.