3D in vitro Model of Vascular Medial Thickening in Pulmonary Arterial

k l FE . CD

h h

A

kA3:l e t f o g

t y - y f t y p gA3:

kA3C 5l t q p g

x u q tf RV R Z u

t o fx y pg

f A3: s A3C 5 6 x fx

x gy fA3: rp q

p kA698 44l f q x t f

x g q fx 6 A3: fA698

f y p t y x s f

t p x t u x g fA3:

f f u x s f

t y g

i

f f f f f f

A

kA3:l e t f o f

t y f - y f t y p g

f p rp x x t y p g A3C 5

f q r f y

p tf fRV R Z t p f t o

g f f p x t y p A3:

A3C 5 fA3: x g

A

A3: s f o p g f

o p s g s A3C 5 h gx A3C 5

f f)+

( o ,% ( , f g 6 A3: A698

44 qfCHD G 9ZNNV TNL T Z AZ KN D NZU 8R NZ CLRNV RORL p

pf k RS V 5) L VO L T T NZ URLZ L XNl p I LS f ;C 7TNUNV 3B NZ R V +% O ZN RS V p g

49

f R-. 3KL U CHD G 9ZNNV f g;V R 5NTT 6N 6N NL R V R B L N p gy f 6 A3:

s B 3 f55 6( 35D4 p BD YA5B pf

g f9Z X A M AZR U / 9Z X A M C O ZN p g A

( 6 A3:

)+ ( o ,% ( , A3:

A3C 5 x f+m- ) dU 6 A3: t u

g

) A698 44 6 A3:

A698 44 fA3: t px t y r f fy z A3:

p fA3: p x t y p o gx A698 44

6 A3: q x f R-. t t g

6 A3:

) 6 A3: t s f

fA698 o q g 6 A3:

t f o 55 6( UB 3 t g f R-.

t f o DE 7 t p g

+ A3: 6 A3:

p f A3: y p tf 6 A3:

to s p gA3: o tf (

g o f o

B7 )-0f n o p gp

6 A3: g f t

vfx 55 6( UB 3 y g f R-.

f DE 7 t g

B

f p rp x x t y p g A3C 5

f q r f y p g

A3: f f p

x t y p A3: A3C 5 p x s fA3:

o q g x t y p A698 44

t fy w vf y p

t y x s f f

u x t y g

p fx s fRV R Z A3:

x gx f u A3: u

to g

50

A (%

ZRR 5 D V S :H ;b R H S H U U K Z : V B P 3% 6 RV R Z MNT O F L T Z NMR T D RLSNVRVP RV A TU V Za 3Z NZR T :aXNZ NV R V% 8Z V 4R NVP 4R NL V T ) ) 2/1+/)%

51

fbioe-08-00482 May 19, 2020 Time: 16:35 # 1

BRIEF RESEARCH REPORT published: 20 May 2020 doi: 10.3389/fbioe.2020.00482

Edited by:

Paul Andrew Wieringa, Maastricht University, Netherlands Reviewed by:

Patricia Diaz-Rodriguez, University of La Laguna, Spain Fatemeh Kabirian, Materials and Energy Research Center, Iran Emilie Velot, Université de Lorraine, France

*Correspondence:

Aiko Ogawa [email protected]

†These authors have equally contributed to the work

Specialty section:

This article was submitted to Biomaterials, a section of the journal Frontiers in Bioengineering and Biotechnology Received:31 January 2020 Accepted:27 April 2020 Published:20 May 2020 Citation:

Morii C, Tanaka HY, Izushi Y, Nakao N, Yamamoto M, Matsubara H, Kano MR and Ogawa A (2020) 3D in vitro Model of Vascular Medial Thickening in Pulmonary Arterial Hypertension.

Front. Bioeng. Biotechnol. 8:482.

doi: 10.3389/fbioe.2020.00482

3D in vitro Model of Vascular Medial Thickening in Pulmonary Arterial

Hypertension

Chiharu Morii^1,2†, Hiroyoshi Y. Tanaka^1†, Yasuhisa Izushi², Natsumi Nakao¹,

Masaya Yamamoto^3,4, Hiromi Matsubara², Mitsunobu R. Kano^1,5and Aiko Ogawa²*

1Department of Pharmaceutical Biomedicine, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University, Okayama, Japan,²Division of Molecular and Cellular Medicine, Department of Clinical Science, National Hospital Organization Okayama Medical Center, Okayama, Japan,³Department of Materials Processing, Graduate School of Engineering, Tohoku University, Sendai, Japan,⁴Graduate School of Biomedical Engineering, Tohoku University, Sendai, Japan,⁵Department of Pharmaceutical Biomedicine, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, Japan

In pulmonary arterial hypertension (PAH), excessive proliferation of pulmonary artery smooth muscle cells (PASMCs) causes vascular medial thickening. Medial thickening is a histopathological hallmark of pulmonary vascular remodeling, the central disease process driving PAH progression. Pulmonary vascular remodeling causes stenosis and/or obstruction of small pulmonary arteries. This leads to increased pulmonary vascular resistance, elevated pulmonary arterial pressure, and ultimately right heart failure. To improve the survival of PAH patients, which remains at approximately 60%

at 3 years after diagnosis, the development of novel PAH-targeted drugs is desired.

To this end, a detailed understanding of the mechanisms underlying excessive PASMC proliferation and the medial thickening that ensues is necessary. However, a lack of in vitromodels that recapitulate medial thickening impedes our deeper understanding of the pathogenetic mechanisms involved. In the present study, we applied 3-dimensional (3D) cell culture technology to develop a novelin vitro model of the pulmonary artery medial layer using human PAH patient-derived PASMCs. The addition of platelet-derived growth factor (PDGF)-BB, a mitogen known to promote excessive PASMC proliferation in PAH, resulted in increased thickness of the 3D-PAH media tissues. Conversely, administration of the PDGF receptor inhibitor imatinib or other clinical PAH drugs inhibited this medial thickening-inducing effect of PDGF-BB. Altogether, by using 3D cell culture technology, we report the generation of anin vitromodel of medial thickening in PAH, which had hitherto not been successfully modeledin vitro. This model is potentially useful for assessing the ability of candidate PAH drugs to suppress medial thickening.

Keywords: pulmonary arterial hypertension, medial thickening, pulmonary artery smooth muscle cell, 3D culture, PDGF signaling, imatinib

INTRODUCTION

Pulmonary arterial hypertension (PAH) is a devastating disease. In PAH, pulmonary vascular remodeling causes stenosis and/or obstruction of small pulmonary arteries. This leads to increased pulmonary vascular resistance, elevated pulmonary arterial pressure, and ultimately right heart failure. The survival rate is approximately 60% at 3 years after diagnosis despite decades of

Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 1 May 2020 | Volume 8 | Article 482

52