Pericytes Elicit Resistance to Vemurafenib and Sorafenib Therapy in Thyroid Carcinoma via the TSP-1/TGFβ1 Axis.

Abstract

The BRAF^V600E oncogene modulates the papillary thyroid carcinoma (PTC) microenvironment, in which pericytes are critical regulators of tyrosine-kinase (TK)-dependent signaling pathways. Although BRAF^V600E and TK inhibitors are available, their efficacy as bimodal therapeutic agents in BRAF^V600E-PTC is still unknown. We assessed the effects of vemurafenib (BRAF^V600E inhibitor) and sorafenib (TKI) as single agents or in combination in BRAF^WT/V600E-PTC and BRAF^WT/WT cells using cell-autonomous, pericyte coculture, and an orthotopic mouse model. We also used BRAF^WT/V600E-PTC and BRAF^WT/WT-PTC clinical samples to identify differentially expressed genes fundamental to tumor microenvironment. Combined therapy blocks tumor cell proliferation, increases cell death, and decreases motility via BRAF^V600E inhibition in thyroid tumor cells <i>in vitro</i>. Vemurafenib produces cytostatic effects in orthotopic tumors, whereas combined therapy (likely reflecting sorafenib activity) generates biological fluctuations with tumor inhibition alternating with tumor growth. We demonstrate that pericytes secrete TSP-1 and TGFβ1, and induce the rebound of pERK1/2, pAKT and pSMAD3 levels to overcome the inhibitory effects of the targeted therapy in PTC cells. This leads to increased BRAF^V600E-PTC cell survival and cell death refractoriness. We find that BRAF^WT/V600E-PTC clinical samples are enriched in pericytes, and TSP1 and TGFβ1 expression evoke gene-regulatory networks and pathways (TGFβ signaling, metastasis, tumor growth, tumor microenvironment/ECM remodeling functions, inflammation, VEGF ligand-VEGF receptor interactions, immune modulation, etc.) in the microenvironment essential for BRAF^WT/V600E-PTC cell survival. Critically, antagonism of the TSP-1/TGFβ1 axis reduces tumor cell growth and overcomes drug resistance. Pericytes shield BRAF^V600E-PTC cells from targeted therapy via TSP-1 and TGFβ1, suggesting this axis as a new therapeutic target for overcoming resistance to BRAF^V600E and TK inhibitors.