Dapagliflozin Inhibits Cell Adhesion to Collagen I and IV and Increases Ectodomain Proteolytic Cleavage of DDR1 by Increasing ADAM10 Activity.

Junichi Okada,Masanobu Yamada,Jeffrey E Pessin,Aya Osaki,Yoko Shimoda,Atsushi Ozawa,Tsugumichi Saito,Shuichi Okada,Hideaki Yokoo,Yasuyo Nakajima,Eijiro Yamada

doi:10.3390/molecules25030495

Junichi Okada, Masanobu Yamada + Show 9 more

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https://doi.org/10.3390/molecules25030495

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Abstract

Dapagliflozin, empagliflozin, tofogliflozin, selective inhibitors of sodium-glucose cotransporter 2 (SGLT2), is used clinically to reduce circulation glucose levels in patients with type 2 diabetes mellitus by blocking the reabsorption of glucose by the kidneys. Dapagliflozin is metabolized and inactivated by UGT1A9. Empagliflozin is metabolized and inactivated by UGT1A9 and by other related isoforms UGT2B7, UGT1A3, and UGT1A8. Tofogliflozin is metabolized and inactivated by five different enzymes CYP2C18, CYP3A4, CYP3A5, CYP4A11, and CYP4F3. Dapagliflozin treatment of HCT116 cells, which express SGLT2 but not UGT1A9, results in the loss of cell adhesion, whereas HepG2 cells, which express both SGLT2 and UGT1A9, are resistant to the adhesion-related effects of dapagliflozin. PANC-1 and H1792 cells, which do not express either SGLT2 or UGT1A9, are also resistant to adhesion related effects of dapagliflozin. On the other hand, either empagliflozin or tofogliflozin treatment of HCT116, HepG2, PANC-1, and H1792 cells are resistant to the adhesion-related effects as observed in dapagliflozin treated HCT116 cells. Knockdown of UGT1A9 by shRNA in HepG2 cells increased dapagliflozin sensitivity, whereas the overexpression of UGT1A9 in HCT116 cells protected against dapagliflozin-dependent loos of cell adhesion. Dapagliflozin treatment had no effect on cellular interactions with fibronectin, vitronectin, or laminin, but it induced a loss of interaction with collagen I and IV. In parallel, dapagliflozin treatment reduced protein levels of the full-length discoidin domain receptor I (DDR1), concomitant with appearance of DDR1 cleavage products and ectodomain shedding of DDR1. In line with these observations, unmetabolized dapagliflozin increased ADAM10 activity. Dapagliflozin treatment also significantly reduced Y792 tyrosine phosphorylation of DDR1 leading to decrement of DDR1 function and detachment of cancer cells. Concomitant with these lines of results, we experienced that CEA in patients with colon cancer, which express SGLT2 but not UGT1A9, and type 2 diabetes mellitus treated by dapagliflozin in addition to chemotherapy was decreased (case 1). CEA in patients with colon cancer, which express SGLT2 but not UGT1A9, and type 2 diabetes mellitus was treated by dapagliflozin alone after radiation therapy was decreased but started to rise after cessation of dapagliflozin (case 2). CA19-9 in two of patients with pancreatic cancer and type 2 diabetes mellitus was resistant to the combination therapy of dapagliflozin and chemotherapy (case 3 and 4 respectively). PIVKAII in patients with liver cancer and type 2 diabetes mellitus, and CYFRA in patients with squamous lung cancer and type 2 diabetes mellitus was also resistant the combination therapy of dapagliflozin and chemotherapy (case 5 and 6 respectively). Taken together, these data suggest a potential role for dapagliflozin anticancer therapy against colon cancer cells that express SGLT2, but not UGT1A9.

Highlights

IntroductionThe first family contains glucose transporter (GLUT) proteins, which are stereospecific facilitative glucose transporters, with net transport driven by the concentration gradient of glucose across the membrane [1]
In mammals, there are two major families of glucose transporters
After 35 min of incubation, HCT116 cells treated with 0.125 mM dapagliflozin began to display morphological changes associated with cell detachment (Figure 1b, left panel), whereas cells treated with 0.5 mM dapagliflozin were almost completely detached from the substratum (Figure 1b, left panel)

Summary

Introduction

The first family contains glucose transporter (GLUT) proteins, which are stereospecific facilitative glucose transporters, with net transport driven by the concentration gradient of glucose across the membrane [1]. The second family contains sodium-dependent glucose cotransporters (SGLTs) [2], which use the sodium concentration gradient to drive the uptake of glucose, allowing for more efficient uptake and net flux against the glucose concentration gradient [2]. It is well established that tumor cells typically undergo a metabolic switch towards utilization of glucose and away from fatty acid oxidation [7]. Cancer cells express many types of GLUTs to maintain the high levels of glucose needed for their high metabolic rates and to provide critical substrates for DNA and RNA synthesis that are not available through fatty acid oxidation [2]

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