Transcriptomic and Functional Evidence for Differential Effects of MCF-7 Breast Cancer Cell-Secretome on Vascular and Lymphatic Endothelial Cell Growth

Abstract

Vascular and lymphatic vessels drive breast cancer (BC) growth and metastasis. We assessed the cell growth (proliferation, migration, and capillary formation), gene-, and protein-expression profiles of Vascular Endothelial Cells (VECs) and Lymphatic Endothelial Cells (LECs) exposed to a conditioned medium (CM) from estrogen receptor-positive BC cells (MCF-7) in the presence or absence of Estradiol. We demonstrated that MCF-7-CM stimulated growth and capillary formation in VECs but inhibited LEC growth. Consistently, MCF-7-CM induced ERK1/2 and Akt phosphorylation in VECs and inhibited them in LECs. Gene expression analysis revealed that the LECs were overall (≈10-fold) more sensitive to MCF-7-CM exposure than VECs. Growth/angiogenesis and cell cycle pathways were upregulated in VECs but downregulated in LECs. An angiogenesis proteome array confirmed the upregulation of 23 pro-angiogenesis proteins in VECs. In LECs, the expression of genes related to ATP synthesis and the ATP content were reduced by MCF-7-CM, whereas MTHFD2, an immune evasion gene, was upregulated. The contrasting effect of MCF-7-CM on the growth of VECs and LECs was reversed by inhibiting the TGF-β signaling pathway. The effect of MCF-7-CM on VEC growth was also reversed by inhibiting the VEGF signaling pathway. In conclusion, BC secretome may facilitate cancer cell survival and tumor growth by simultaneously promoting vascular angiogenesis and inhibiting lymphatic growth. The differential effects of BC secretome on LECs and VECs may be of pathophysiological relevance in BC.

Transcriptomic and Functional Evidence That miRNA193a-3p Inhibits Lymphatic Endothelial Cell (LEC) and LEC + MCF-7 Spheroid Growth Directly and by Altering MCF-7 Secretome

“…There are several limitations of the present study that should be addressed in the future. Our observation that in LECs, miR193a-3p inhibits growth effects of MCF-7 secretome, which in-itself attenuates LEC growth [52], suggests that miR193a3p enhances the growth attenuating actions of MCF-7 secretome. Hence, the inhibitory actions of miR193a-3p on LEC growth should be confirmed using secretomes from BC cells, which stimulate LEC growth, i.e.…”

“…Moreover, BC-derived factors can educate LECs to promote BC cell growth in a paracrine fashion [5]. Interestingly, MCF-7 secretome also induces proliferation, migration, and capillary formation in VECs [29,52]. Moreover, LECderived factors educate BC cells to produce/secrete more growth-promoting molecules in their secretome to enhance tumor growth, as well as increased angiogenesis [51].…”

Selective refueling of CAR T cells using ADA1 and CD26 boosts antitumor immunity

“…Third, our strategy overexpresses CD26 on the surface of CAR T cells, as the CD26 expression level on the unmodified CAR T cells is relatively low and can be further downregulated by TGF-β1, a cytokine found in significant quantities in various tumor tissues. 68 , 69 , 70 , 71 , 72 , 73 , 74 Thus, we develop a strategy to overexpress CD26 on the surface of CAR T cells, which sustains co-stimulation and membrane-proximal ADA1 capture even in TGF-β1-rich environment and provides a survival advantage for these engineered therapeutic cells.…”

Selective refueling of CAR T cells using ADA1 and CD26 boosts antitumor immunity

“…Cytoplasmic ADA1 converts intracellular adenosine to inosine, and upon secretion, produces inosine in a CAR T cell membrane-proximal manner, which avoids feeding tumor cells. Third, our strategy overexpresses CD26 on the surface of CAR T cells as the CD26 expression level on the unmodified CAR T cells is relatively low and can be further downregulated by TGF-ß1, a cytokine found in significant quantities in various tumor tissues [72][73][74][75][76][77][78] . Thus, we develop a strategy to overexpress CD26 on the surface of CAR T cells, which sustains co-stimulation and membrane proximal ADA1 capture even in TGF-ß1-rich environment and provides a survival advantage for these engineered therapeutic cells.…”