HIF2A–LOX Pathway Promotes Fibrotic Tissue Remodeling in Thyroid-Associated Orbitopathy

Hikage, Fumihito; Atkins, Stephen; Kahana, Alon; Smith, Terry J.; Chun, Tae Hwa

doi:10.1210/en.2018-00272

Cited by 73 publications

(145 citation statements)

References 53 publications

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“…Furthermore, these 3D culture techniques may have some limitations in terms of mimicking the physiological and pathological conditions because of the presence of an unnecessary scaffolds. Alternatively, our group recently developed a 3D cell drop culture system in which no scaffold is required 25 – 27 . Using this technique, we were able to obtain 3D organoids from HOF in patients with TAO using a drop culture method, and found that HIF2A played a critical role in mediating LOX-dependent ECM accumulation 25 .…”

Section: Discussionmentioning

confidence: 99%

“…Alternatively, our group recently developed a 3D cell drop culture system in which no scaffold is required 25 – 27 . Using this technique, we were able to obtain 3D organoids from HOF in patients with TAO using a drop culture method, and found that HIF2A played a critical role in mediating LOX-dependent ECM accumulation 25 . More recently, to elucidate the molecular etiology of DUES induced by PGs, we employed this 3D culture system using HOF and 3T3-L1 cells, and found that PGs significantly suppressed 3D organoid sizes, and modulated the spatial distribution of several ECMs surrounding the 3D organoids 26 , 27 .…”

Section: Discussionmentioning

confidence: 99%

“…The 3D organoids of HTM were generated by a hanging droplet spheroid three-dimension (3D) culture system as described in a previous report 25 . Briefly, 90% confluence HTM cells in 150 mm 2D culture dishes as above were washed with phosphate buffered saline (PBS), and the cells were detached using 0.25% Trypsin/EDTA.…”

Section: Methodsmentioning

confidence: 99%

“…Although most of these studies involved the use of conventional 2D cell cultures, despite the fact that the HTM is composed of multiple sheets in the human eye 24 , thus a relevant 3D cell culture model should be used in this research field. Our group recently established a 3D cell culture system using 3T3-L1 cells or human orbital fibroblasts (HOF) as disease models for thyroid associated orbitopathy (TAO) 25 , and the deepening of the upper eyelid sulcus (DUES) induced by prostaglandin analogues (PGs) 26 , 27 .…”

Section: Introductionmentioning

confidence: 99%

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ROCK inhibitors beneficially alter the spatial configuration of TGFβ2-treated 3D organoids from a human trabecular meshwork (HTM)

Ota

Ida

Ohguro

et al. 2020

Sci Rep

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101

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To elucidate molecular pharmacology of Rho-associated coiled-coil containing protein kinase inhibitors (ROCK-i, Ripasudil and Y27632) on their efficiency for aqueous outflow, 2D or 3D cultures of a human trabecular meshwork (HTM) were prepared in the presence of TGFβ2. Those were examined by transendothelial electrical resistance (TEER, 2D), electronic microscopy (EM, 2D and 3D), expression of the extracellular matrix (ECM) including collagen1 (COL1), COL4 and COL6, and fibronectin (FN) by immunolabeling and/or quantitative PCR (3D), and solidity of 3D organoids by a micro-squeezer. TGFβ2 significantly increased the TEER values in 2D cultures, and the ECM expression indicated that the 3D organoids assumed a more densely packed shape. ROCK-i greatly reduced the TGFβ2-induced enhancement of TEER and the immunolabeled ECM expression of the 3D organoids. In contrast, the mRNA expression of COL1 was increased, and those of COL4 and FN were unchanged. EM revealed that TGFβ2 caused the HTM cells to become more compact and abundant ECM deposits within the 3D organoids were observed. These were significantly inhibited by ROCK-i. The dense solids caused by the presence of TGFβ2 were significantly suppressed by ROCK-i. Current study indicates that ROCK-i cause beneficial effects toward the spatial configuration of TGFβ2-induced HTM 3D organoids.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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ROCK inhibitors beneficially alter the spatial configuration of TGFβ2-treated 3D organoids from a human trabecular meshwork (HTM)

Ota

Ida

Ohguro

et al. 2020

Sci Rep

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101

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“…28 Some studies have reported that cell behavior is altered notably when cells are embedded within a 3D matrix in contrast with the cell-matrix interactions seen in 2D cultures. 29,30 Thus, the 3D tissue culture system has emerged as a novel ex vivo approach to modeling many diseases, 31,32 and the recently developed 3D organoid culture system appears to be more suitable for identifying the mechanisms within adipogenesis that are involved in the etiology of DUES caused by PG analogs. In addition, several studies have suggested that physiological properties of orbital fatty tissues may be different from those of 3T3-L1 cells.…”

mentioning

confidence: 99%

Prostaglandin F2α Agonists Negatively Modulate the Size of 3D Organoids from Primary Human Orbital Fibroblasts

Itoh

Hikage

Ida

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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108

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To elucidate the molecular etiology of deepening of the upper eyelid sulcus (DUES) induced by prostaglandin (PG) analogs, a three-dimensional (3D) tissue culture system was employed using human orbital fibroblasts (HOFs). METHODS. During adipogenesis, changes in HOF 3D organoid sizes, as well as their lipids stained by BODIPY and expression of the extracellular matrix (ECM) by immunolabeling and/or quantitative PCR, were studied in the presence or absence of either 100-nM bimatoprost acid or 100-nM prostaglandin F2α. RESULTS. The size of the 3D organoids increased remarkably during adipogenesis, but such increases were significantly inhibited by the presence of PG analogs. Staining intensities by BODIPY and mRNA expression of peroxisome proliferator-activated receptor gamma were significantly increased upon adipogenesis but were not influenced by the presence of PG analogs. Unique changes in ECM expression observed with or without adipogenic differentiation were significantly modified by the presence of PG analogs. CONCLUSIONS. Our present study indicates that PG analogs have the potential to modulate the ECM network within HOF 3D organoids. Thus, a 3D tissue culture system may be a suitable strategy for understanding the disease etiology of DUES.

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Pathogenesis and Medical Management of Thyroid Eye Disease

Carniciu¹,

Kahana²

2020

Smith and Nesi’s Ophthalmic Plastic and Reconstructive Surgery

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HIF2A–LOX Pathway Promotes Fibrotic Tissue Remodeling in Thyroid-Associated Orbitopathy

Cited by 73 publications

References 53 publications

ROCK inhibitors beneficially alter the spatial configuration of TGFβ2-treated 3D organoids from a human trabecular meshwork (HTM)

ROCK inhibitors beneficially alter the spatial configuration of TGFβ2-treated 3D organoids from a human trabecular meshwork (HTM)

Prostaglandin F2α Agonists Negatively Modulate the Size of 3D Organoids from Primary Human Orbital Fibroblasts

Pathogenesis and Medical Management of Thyroid Eye Disease

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