Cytochrome P450 Activity in Ex Vivo Cornea Models and a Human Cornea Construct

Kölln, Christian; Reichl, Stephan

doi:10.1016/j.xphs.2016.04.010

Cited by 9 publications

(4 citation statements)

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“…17 In our hands, this treatment reliably induces Cre not only in the eye, but also in tissues throughout the body including the kidney, presumably due to its metabolization by corneal CYP2D6 (a cytochrome P450 isoform) and uptake from the ocular surface into the general circulation. 18,19 Successful induction was first demonstrated by crossing CAGG-Cre-ER TM mice with mT/mG mice, a double-fluorescent Cre reporter mouse strain that expresses membrane-targeted green fluorescent protein (mG) after excision, reflecting activation of the Cre recombinase. 11 To this end the experimental animals were treated from P1 to P5 three times daily with tamoxifen eye drops (2.5 mg/ml) and the isolated tissue was examined for fluorescence at 5-6 weeks of age.…”

Section: Generation Of Newborn Mice With Induced Fn Deficiencymentioning

confidence: 99%

Fibronectin deficiency in newborn mice leads to cyst formation in the kidney

Hermann

Seibold²,

Skoczynski

et al. 2022

Preprint

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PurposeThe ubiquitously expressed glycoprotein fibronectin (FN) is a central component of the fibrillar extracellular matrix (ECM) that is found in multiple sites throughout the body including the peritubular interstitium of the kidney. To learn more about the specific role(s) of FN in the kidney we generated and investigated FN-deficient mice.MethodsWe generated CAGG-Cre-ER™/Fnfl/fl mice which carry floxed Fn alleles and ubiquitously express Cre-recombinase after tamoxifen treatment. Newborn pups were treated with tamoxifen eye drops (2.5 mg/mL) to induce FN deficiency. Conditional deletion of Fn was confirmed by quantitative real-time PCR, Western blot analysis and immunohistochemistry. The expression patterns of Fn were analyzed by in situ hybridization. Kidneys were investigated by light microscopy and immunohistochemistry.ResultsThe expression analyses and immunohistochemistry showed a significant reduction of FN at postnatal day (P) 4. Loss of FN corelated with the formation of renal cysts at the corticomedullary border, which expand with increasing age. In situ hybridization demonstrated that on P4 Fn expression extends mainly from the pelvis to the corticomedullary border, whereas in 5-6 weeks old mice it is located only in the cortex. Immunohistochemistry and light microscopy showed a loosening of the renal interstitium and additionally an appearance of ECM proteins in the cysts.ConclusionWe conclude that FN deficiency leads to the development of renal cysts, which occurs a few days after tamoxifen treatment and results in extensive loss of renal parenchyma a few weeks after birth. The results indicate an important role of FN for maintenance of kidney structure and function.

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Section: Generation Of Newborn Mice With Induced Fn Deficiencymentioning

confidence: 99%

Fibronectin deficiency in newborn mice leads to cyst formation in the kidney

Hermann

Seibold²,

Skoczynski

et al. 2022

Preprint

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“…The cells might interact even through the PC membrane as the pore size of 3 µm is much wider than the molecule size of cytokines and other signal molecules. For this reason, the benefit of co-cultivation remains unchanged and, thus, other cell characteristics, such as substance transport [52][53][54] and substance metabolism [27,28], which we investigated extensively in our former studies, might also be maintained. This finding was also supported by the cell viability results.…”

Section: Adaption Of Hc Cultivationmentioning

confidence: 99%

DynaMiTES – A dynamic cell culture platform for in vitro drug testing PART 2 – Ocular DynaMiTES for drug absorption studies of the anterior eye

Beiβner

Mattern

Dietzel

et al. 2018

European Journal of Pharmaceutics and Biopharmaceutics

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In the present study, a formerly designed Dynamic Micro Tissue Engineering System (DynaMiTES) was applied with our prevalidated human hemicornea (HC) construct to obtain a test platform for improved absorption studies of the anterior eye (Ocular DynaMiTES). First, the cultivation procedure of the classic HC was slightly adapted to the novel DynaMiTES design. The obtained inverted HC was then compared to classic HC regarding cell morphology using light and scanning electron microscopy, cell viability using MTT dye reaction and epithelial barrier properties observing transepithelial electrical resistance and apparent permeation coefficient of sodium fluorescein. These tested cell criteria were similar. In addition, the effects of four different flow rates on the same cell characteristics were investigated using the DynaMiTES. Because no harmful potential of flow was found, dynamic absorption studies of sodium fluorescein with and without 0.005%, 0.01% and 0.02% benzalkonium chloride were performed compared to the common static test procedure. In this proof-of-concept study, the dynamic test conditions showed different results than the static test conditions with a better prediction of in vivo data. Thus, we propose that our DynaMiTES platform provides great opportunities for the improvement of common in vitro drug testing procedures.

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“…Notably, some of these topical medications (substrates, inducers, or inhibitors of P450s) are known to elicit both ocular and systemic effects ( Syed et al, 2021 ). For example, among the drugs for glaucoma treatment, a major cause of legal blindness ( Quigley and Broman, 2006 ), these are timolol metabolized by CYP2D6; betaxolol, a substrate for CYP1A2 and CYP2D6; dorzolamide eliminated by CYP2B1, CYP2C9, CYP2E1, and CYP3A2; and pilocarpine, which inhibits CYP2A6, CYP2A13, and CYP2E1 ( Kimonen et al, 1995 ). Particular caution is recommended when ophthalmic timolol or betaxolol are coadministered with paroxetine or other strong CYP2D6 inhibitors ( Farkouh et al, 2016 ; Vaajanen and Vapaatalo, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Challenges and Opportunities in P450 Research on the Eye

Pikuleva

2023

Drug Metab Dispos

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Of the 57 cytochrome P450 enzymes found in humans, at least 30 have ocular tissues as an expression site. Yet knowledge of the roles of these P450s in the eye is limited, in part because only very few P450 laboratories expanded their research interests to studies of the eye. Hence the goal of this review is to bring attention of the P450 community to the eye and encourage more ocular studies. This review is also intended to be educational for eye researchers and encourage their collaborations with P450 experts. The review starts with a description of the eye, a fascinating sensory organ, and is followed by sections on ocular P450 localizations, specifics of drug delivery to the eye, and individual P450s, which are grouped and presented based on their substrate preferences. In sections describing individual P450s, available eye-relevant information is summarized and concluded by the suggestions on the opportunities in ocular studies of the discussed enzymes. Potential challenges are addressed as well. The conclusion section outlines several practical suggestions on how to initiate eye-related research. SIGNIFICANCE STATEMENT This review focuses on the cytochrome P450 enzymes in the eye to encourage their ocular investigations and collaborations between P450 and eye researchers.

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Cytochrome P450 Activity in Ex Vivo Cornea Models and a Human Cornea Construct

Cited by 9 publications

References 50 publications

Fibronectin deficiency in newborn mice leads to cyst formation in the kidney

Fibronectin deficiency in newborn mice leads to cyst formation in the kidney

DynaMiTES – A dynamic cell culture platform for in vitro drug testing PART 2 – Ocular DynaMiTES for drug absorption studies of the anterior eye

Challenges and Opportunities in P450 Research on the Eye

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