Inhibition of pressure-activated cancer cell adhesion by FAK-derived peptides

Abstract

Forces within the surgical milieu or circulation activate cancer cell adhesion and potentiate metastasis through signaling requiring FAK-Akt1 interaction. Impeding FAK-Akt1 interaction might inhibit perioperative tumor dissemination, facilitating curative cancer surgery without global FAK or AKT inhibitor toxicity. Serial truncation and structurally designed mutants of FAK identified a seven amino acid, short helical structure within FAK that effectively competes with Akt1-FAK interaction. Adenoviral overexpression of this FAK-derived peptide inhibited pressure-induced FAK phosphorylation and AKT-FAK coimmunoprecipitation in human SW620 colon cancer cells briefly exposed to 15mmHg increased pressure, consistent with laparoscopic or post-surgical pressures. Adenoviral FAK-derived peptide expression prevented pressure-activation of SW620 adhesion not only to collagen-I-coated plates but also to murine surgical wounds. A scrambled peptide did not. Finally, we modeled operative shedding of tumor cells before irrigation and closure by transient cancer cell adhesion to murine surgical wounds before irrigation and closure. Thirty minute preincubation of SW620 cells at 15mmHg increased pressure impaired subsequent tumor free survival in mice exposed to cells expressing the scrambled peptide. The FAK-derived sequence prevented this. These results suggest that blocking FAK-Akt1 interaction may prevent perioperative tumor dissemination and that analogs or mimics of this 7 amino acid FAK-derived peptide could impair metastasis.

ZINC4085554 inhibits cancer cell adhesion by interfering with the interaction of Akt1 and FAK

“…Extracellular pressure treatment. Cells (50,000 cells/well) were seeded into 24 well plates that were then subjected to 15 mmHg increased extracellular pressure for 30 min at 37˚C in a mixture of 5% CO 2 with 95% room air in a pre-warmed air tight box with inlet and outlet valves and a pressure gauge, as described previously (17,24,27). It has been previously reported that this technique, if the box is appropriately pre-warmed, allows for the control temperature, pressure and oxygen partial pressure within the cell culture medium.…”

“…Adhesion assay. Our previous studies performed cell counting via optical microscope (23,28) and Tag-it dye (Tag-it Violet Proliferation Cell Tracking Dye; Biolegend, Inc., San Diego, CA, USA) staining method (24,27) for cell counting and have revealed that dye staining methods are more objective as they do not require the observer to select which microscopic fields to count and are more accurate as they compensate for any unevenness of distribution of cells across the well. Therefore, the adherent cells were not imaged, and only their optical densities were measured.…”

“…Serial truncation of the FAK FERM domain demonstrated that FAK-Akt1 binding occurs in sub domain 1-126 amino acids (22). Further serial truncation of this domain narrowed down to 33 amino acids NT1-2-2 FERM (amino acid sequence 94-126 of FERM) subdomain of FAK is required for Akt1 binding (23), and further studies condensed this further to the 7 amino acid sequence 113LAHPPEEQ119, which is a short helical structure (24). Adenoviral expression of this peptide in colon cancer cells prevents activation of FAK in response to extracellular pressure, attenuates FAK-Akt1 interaction, and inhibits the pressure-induced adhesion of colon cancer cells to a collagen I substrate (24).…”

“…Further serial truncation of this domain narrowed down to 33 amino acids NT1-2-2 FERM (amino acid sequence 94-126 of FERM) subdomain of FAK is required for Akt1 binding (23), and further studies condensed this further to the 7 amino acid sequence 113LAHPPEEQ119, which is a short helical structure (24). Adenoviral expression of this peptide in colon cancer cells prevents activation of FAK in response to extracellular pressure, attenuates FAK-Akt1 interaction, and inhibits the pressure-induced adhesion of colon cancer cells to a collagen I substrate (24). Furthermore, cells transiently overexpressing this peptide are less adherent in murine surgical wound tumor recurrence models, decreasing tumor incidence and improving subsequent survival (24).…”

“…Adenoviral expression of this peptide in colon cancer cells prevents activation of FAK in response to extracellular pressure, attenuates FAK-Akt1 interaction, and inhibits the pressure-induced adhesion of colon cancer cells to a collagen I substrate (24). Furthermore, cells transiently overexpressing this peptide are less adherent in murine surgical wound tumor recurrence models, decreasing tumor incidence and improving subsequent survival (24).…”