Introduction: It has been demonstrated that exposure of endothelial cells to hypercholesterolemia and hyperglycemia leads to increased levels of endoglin and cell adhesion molecules, as well as increased adhesion and transmigration of monocytes through endothelial monolayer. Carotuximab (TRC105), a monoclonal antibody that binds to endoglin, was originally developed for the use in oncology. However, there are no studies available to elucidate potential role of carotuximab treatment on Eng expression, signaling and function with respect to endothelial dysfunction development and/or prevention. Aim: Therefore, in this study, we hypothesized that carotuximab treatment prevents 7-ketocholesterol and hyperglycemia induced development of endothelial dysfunction by direct effect on Eng expression, signaling and function. Material and Methods: Human aortic endothelial cells (HAECs), passage 5, were cultured in EGM-2 media with appropriate supplements and 10% FBS until reaching 80% confluence. In hypercholesterolemia studies, cells were treated with carotuximab (300 μg/mL) for 1 hour, followed by addition of 7-ketocholesterol (10 μg/mL) for another 12 h. In hyperglycemia studies, cells were exposed to high glucose (45 mM) for 60 hours, followed by addition of carotuximab for another 12 hours and cells treated with 5 mM glucose and 40 mM mannitol served as osmotic control. Gene expression was measured by qRT-PCR. Protein levels, adhesion and transmigration of monocytes were assessed by flow cytometry. Results: Carotuximab pretreatment reduced endoglin protein expression and signaling in both hypercholesterolemia and hyperglycemia induced endothelial dysfunction. Despite increased expression of cell adhesion molecules carotuximab blockage of endoglin prevented increase of adhesion and transmigration of monocytes through endothelial monolayer in both hypercholesterolemia and hyperglycemia induced endothelial dysfunction. Conclusion: These results suggest that carotuximab-mediated endoglin blockage is essential in hypercholesterolemia and hyperglycemia induced endothelial dysfunction in HAECs and therefore that endoglin might be an interesting therapeutical target in diseases characterized by elevated cholesterol and glucose.

Department of Biological and Medical Sciences Faculty of Pharmacy in Hradec Kralove Charles University

Department of Medical Biochemistry Faculty of Medicine in Hradec Králové Charles University

000      

00000naa a2200000 a 4500

001      

bmc21029346

003      

CZ-PrNML

005      

20220201162706.0

007      

ta

008      

211206s2021 xr f 000 0|eng||

009      

PC

040    __

$a ABA008 $b cze $d ABA008 $e AACR2

041    0_

$a eng

044    __

$a xr

100    1_

$a Tripska, K. $u Department of Biological and Medical Sciences, Faculty of Pharmacy in Hradec Kralove, Charles University

245    10

$a Endoglin blockage is essential in hypercholesterolemia and hyperglycemia induced endothelial dysfunction in HAECs / $c Tripska K, Igreja e Sá IC., Vicen M, Havelek R, Eissazadeh S, Vitverová B, Nachtigal P.

520    9_

$a Introduction: It has been demonstrated that exposure of endothelial cells to hypercholesterolemia and hyperglycemia leads to increased levels of endoglin and cell adhesion molecules, as well as increased adhesion and transmigration of monocytes through endothelial monolayer. Carotuximab (TRC105), a monoclonal antibody that binds to endoglin, was originally developed for the use in oncology. However, there are no studies available to elucidate potential role of carotuximab treatment on Eng expression, signaling and function with respect to endothelial dysfunction development and/or prevention. Aim: Therefore, in this study, we hypothesized that carotuximab treatment prevents 7-ketocholesterol and hyperglycemia induced development of endothelial dysfunction by direct effect on Eng expression, signaling and function. Material and Methods: Human aortic endothelial cells (HAECs), passage 5, were cultured in EGM-2 media with appropriate supplements and 10% FBS until reaching 80% confluence. In hypercholesterolemia studies, cells were treated with carotuximab (300 μg/mL) for 1 hour, followed by addition of 7-ketocholesterol (10 μg/mL) for another 12 h. In hyperglycemia studies, cells were exposed to high glucose (45 mM) for 60 hours, followed by addition of carotuximab for another 12 hours and cells treated with 5 mM glucose and 40 mM mannitol served as osmotic control. Gene expression was measured by qRT-PCR. Protein levels, adhesion and transmigration of monocytes were assessed by flow cytometry. Results: Carotuximab pretreatment reduced endoglin protein expression and signaling in both hypercholesterolemia and hyperglycemia induced endothelial dysfunction. Despite increased expression of cell adhesion molecules carotuximab blockage of endoglin prevented increase of adhesion and transmigration of monocytes through endothelial monolayer in both hypercholesterolemia and hyperglycemia induced endothelial dysfunction. Conclusion: These results suggest that carotuximab-mediated endoglin blockage is essential in hypercholesterolemia and hyperglycemia induced endothelial dysfunction in HAECs and therefore that endoglin might be an interesting therapeutical target in diseases characterized by elevated cholesterol and glucose.

590    __

$a NEINDEXOVÁNO

655    _7

$a abstrakt z konference $7 D016416 $2 czmesh

700    1_

$a Igreja, e Sá I. C. $u Department of Biological and Medical Sciences, Faculty of Pharmacy in Hradec Kralove, Charles University

700    1_

$a Vicen, M. $u Department of Biological and Medical Sciences, Faculty of Pharmacy in Hradec Kralove, Charles University

700    1_

$a Havelek, Radim $u Department of Medical Biochemistry, Faculty of Medicine in Hradec Králové, Charles University $7 xx0230947

700    1_

$a Eissazadeh, S. $u Department of Biological and Medical Sciences, Faculty of Pharmacy in Hradec Kralove, Charles University

700    1_

$a Vitverová, B. $u Department of Biological and Medical Sciences, Faculty of Pharmacy in Hradec Kralove, Charles University

700    1_

$a Nachtigal, Petr $u Department of Biological and Medical Sciences, Faculty of Pharmacy in Hradec Kralove, Charles University $7 uk2009304471

773    0_

$w MED00188266 $t Athero Review $x 2464-6563 $g Roč. 6, Suppl. 2 (2021), s. 23

856    41

$u https://www.prolekare.cz/casopisy/athero-review/2021-supplementum-2-3/56ps-endoglin-blockage-is-essential-in-hypercholesterolemia-and-hyperglycemia-induced-endothelial-dysfunction-in-haecs-128935 $y plný text volně dostupný

910    __

$a ABA008 $b B 2768 $c 401b $y p $z 0

990    __

$a 20211206 $b ABA008

991    __

$a 20220201162657 $b ABA008

999    __

$a ok $b bmc $g 1755225 $s 1149899

BAS    __

$a 6

BAS    __

$a PreBMC

BMC    __

$a 2021 $b 6 $c Suppl. 2 $d 23 $i 2464-6563 $m AtheroReview $x MED00188266 $y 128935

LZP    __

$b NLK111 $a Meditorial-20211206