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Investigating Carcinine Transport and the Expression Profile of Transporter Genes in Human Corneal Epithelial Cells

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Studies on the Cornea and Lens

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Abstract

Oxidative damage is associated with multiple ocular diseases including corneal disorders, cataract, and retinal degeneration. Thus, the possibility to use the multifunctional antioxidant carcinine (β-alanyl-histamine) as a therapeutic to prevent and reverse intracellular oxidative damage in ocular diseases is promising. Today, the major barrier to developing new ophthalmic treatments is the identification and implementation of effective methods of drug delivery in the eye. The aim of this study was to investigate carcinine transport and the expression profile of transporter genes in corneal epithelial cells. These cells form the blood-aqueous barrier of the cornea, preventing the penetration of topical drugs.

The posterior tissue bioavailability and biological effect of carcinine was measured in mouse retina, following topical delivery. The transport and biological effect of carcinine was measured in cultured human corneal epithelial cells (HCECs). The expression profile of 84 transporter genes was determined in HCECs by quantitative RT-PCR.

We found that carcinine efficiently penetrated the eye globe and the retinal cells after topical delivery in mouse eyes. However, carcinine was not transported into the intracellular compartment of the HCECs and the putative carcinine transporters PepT1 and PepT2 had negligible expression levels in these cells. Five other members of the solute carrier (SLC) superfamily of transporters, encoding influx pumps for amino acids, monocarboxylates, and copper ions were highly expressed in HCECs.

We conclude that carcinine does not penetrate the eye globe through the blood-aqueous barrier of the cornea, suggesting a conjunctival/scleral penetration. Identification of transporters SLC3A2, SLC7A5, SLC16A1, SLC31A1, and SLC38A2 as highly expressed in the corneal epithelium may facilitate the design of topical ophthalmic drugs with enhanced intraocular bioavailability.

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Abbreviations

4-HNE:

4-Hydroxynonenal

ABC:

ATP-binding cassette

ATA2:

Amino acid transporter A2

ATP:

Adenosine triphosphase

COPT1:

Copper transporter 1

Ct:

Cycle threshold

HCEC:

Human corneal epithelial cell

HPLC:

High-performance liquid chromatography

HPRT1:

Hypoxanthine phosphoribosyltransferase

LAT1:

Large neutral amino acid transporter 1

MCT1:

Monocarboxylate transporter 1

MRP:

Multidrug resistance-associated protein

MS:

Mass spectrometry

PEPT:

Peptide transporter

PMP70:

Peroxisomal membrane protein of 70 kDA

RT-PCR:

Reverse transcription polymerase chain reaction

SLC:

Solute carrier

VDAC:

Voltage-dependent anion channel

References

  1. Behar-Cohen F. Drug delivery to the eye: current trends and future perspectives. Ther Deliv. 2012;3:1135–7.

    Article  CAS  PubMed  Google Scholar 

  2. Edelhauser HF, Rowe-Rendleman CL, Robinson MR, et al. Ophthalmic drug delivery systems for the treatment of retinal diseases: basic research to clinical applications. Invest Ophthalmol Vis Sci. 2010;51:5403–20.

    Article  PubMed Central  PubMed  Google Scholar 

  3. Lambiase A, Tirassa P, Micera A, Aloe L, Bonini S. Pharmacokinetics of conjunctivally applied nerve growth factor in the retina and optic nerve of adult rats. Invest Ophthalmol Vis Sci. 2005;46:3800–6.

    Article  PubMed  Google Scholar 

  4. Williams KA, Brereton HM, Farrall A, et al. Topically applied antibody fragments penetrate into the back of the rabbit eye. Eye (Lond). 2005;19:910–3.

    Article  CAS  Google Scholar 

  5. Furrer E, Berdugo M, Stella C, et al. Pharmacokinetics and posterior segment biodistribution of ESBA105, an anti-TNF-alpha single-chain antibody, upon topical administration to the rabbit eye. Invest Ophthalmol Vis Sci. 2009;50:771–8.

    Article  PubMed  Google Scholar 

  6. Mitra AK. Role of transporters in ocular drug delivery system. Pharm Res. 2009;26:1192–6.

    Article  CAS  PubMed  Google Scholar 

  7. Babizhayev MA, Guiotto A, Kasus-Jacobi A. N-Acetylcarnosine and histidyl-hydrazide are potent agents for multitargeted ophthalmic therapy of senile cataracts and diabetic ocular complications. J Drug Target. 2009;17:36–63.

    Article  CAS  PubMed  Google Scholar 

  8. Marchette LD, Wang H, Li F, Babizhayev MA, Kasus-Jacobi A. Carcinine has 4-hydroxynonenal scavenging property and neuroprotective effect in mouse retina. Invest Ophthalmol Vis Sci. 2012;53:3572–83.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. Kasus-Jacobi A, Marchette LD, Xu C, Li F, Wang H, Babizhayev MA. Mechanisms of RDH12-induced Leber congenital amaurosis and therapeutic approaches. Adv Ophthalmol. 2012;1:473–96.

    Google Scholar 

  10. Zhang T, Xiang CD, Gale D, Carreiro S, Wu EY, Zhang EY. Drug transporter and cytochrome P450 mRNA expression in human ocular barriers: implications for ocular drug disposition. Drug Metab Dispos. 2008;36:1300–7.

    Article  CAS  PubMed  Google Scholar 

  11. Xiang CD, Batugo M, Gale DC, et al. Characterization of human corneal epithelial cell model as a surrogate for corneal permeability assessment: metabolism and transport. Drug Metab Dispos. 2009;37:992–8.

    Article  CAS  PubMed  Google Scholar 

  12. Geissler S, Zwarg M, Knutter I, Markwardt F, Brandsch M. The bioactive dipeptide anserine is transported by human proton-coupled peptide transporters. FEBS J. 2010;277:790–5.

    Article  CAS  PubMed  Google Scholar 

  13. Araki-Sasaki K, Ohashi Y, Sasabe T, et al. An SV40-immortalized human corneal epithelial cell line and its characterization. Invest Ophthalmol Vis Sci. 1995;36:614–21.

    CAS  PubMed  Google Scholar 

  14. Bonomo RP, D’Alessandro F, Grasso G, et al. Carcinine-beta-cyclodextrin derivatives as scavenger entities of (OH)-O-center dot radicals and SOD-like properties of their copper(II) complexes. Inorg Chim Acta. 2008;361:1705–14.

    Article  CAS  Google Scholar 

  15. Marchette LD, Thompson DA, Kravtsova M, Ngansop TN, Mandal MN, Kasus-Jacobi A. Retinol dehydrogenase 12 detoxifies 4-hydroxynonenal in photoreceptor cells. Free Radic Biol Med. 2010;48:16–25.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  16. Dahlin A, Geier E, Stocker SL, et al. Gene expression profiling of transporters in the solute carrier and ATP-binding cassette superfamilies in human eye substructures. Mol Pharm. 2013;10:650–63.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Pereira HA, Ruan X, Gonzalez ML, Tsyshevskaya-Hoover I, Chodosh J. Modulation of corneal epithelial cell functions by the neutrophil-derived inflammatory mediator CAP37. Invest Ophthalmol Vis Sci. 2004;45:4284–92.

    Article  PubMed  Google Scholar 

  18. Griffith GL, Russell RA, Kasus-Jacobi A, et al. CAP37 activation of PKC promotes human corneal epithelial cell chemotaxis. Invest Ophthalmol Vis Sci. 2013;54:6712–23.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  19. Holland IB. ABC transporters, mechanisms and biology: an overview. Essays Biochem. 2011;50:1–17.

    Article  CAS  PubMed  Google Scholar 

  20. Hediger MA, Clemencon B, Burrier RE, Bruford EA. The ABCs of membrane transporters in health and disease (SLC series): introduction. Mol Aspects Med. 2013;34:95–107.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  21. Wang E, Xi Z, Li Y, et al. Interaction between platinum complexes and the C-terminal motif of human copper transporter 1. Inorg Chem. 2013;52:6153–9.

    Article  CAS  PubMed  Google Scholar 

  22. Chen HH, Kuo MT. Overcoming platinum drug resistance with copper-lowering agents. Anticancer Res. 2013;33:4157–61.

    CAS  PubMed Central  PubMed  Google Scholar 

  23. Cai H, Wu JS, Muzik O, Hsieh JT, Lee RJ, Peng F. Reduced 64Cu uptake and tumor growth inhibition by knockdown of human copper transporter 1 in xenograft mouse model of prostate cancer. J Nucl Med. 2014;55:622–8.

    Article  CAS  PubMed  Google Scholar 

  24. Tsigelny IF, Sharikov Y, Greenberg JP, et al. An all-atom model of the structure of human copper transporter 1. Cell Biochem Biophys. 2012;63:223–34.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  25. Babizhayev MA, Micans P, Guiotto A, Kasus-Jacobi A. N-acetylcarnosine lubricant eyedrops possess all-in-one universal antioxidant protective effects of L-carnosine in aqueous and lipid membrane environments, aldehyde scavenging, and transglycation activities inherent to cataracts: a clinical study of the new vision-saving drug N-acetylcarnosine eyedrop therapy in a database population of over 50,500 patients. Am J Ther. 2009;16:517–33.

    Article  PubMed  Google Scholar 

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Acknowledgements

This work was supported by grants HR10-152 from the Oklahoma Center for the Advancement of Science and Technology and R01EY015534 from the National Institute of Health. We would like to thank the Laboratory for Molecular Biology and Cytometry Research at the University of Oklahoma Health Sciences Center for the HPLC-MS analysis of unlabeled carcinine.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences and Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, 1110 N. Stonewall Avenue, Oklahoma City, OK, 73117-1223, USA

    Anne Kasus-Jacobi

  2. Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Okhil K. Nag & Vibudhuta Awasthi

  3. Innovative Vision Products Inc., New Castle, DE, USA

    Mark A. Babizhayev

  4. Innovative Vision Products Inc., Moscow, Russia

    Mark A. Babizhayev

  5. Departments of Pharmaceutical Sciences, Pathology, and Cell Biology and Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    H. Anne Pereira

Authors
  1. Anne Kasus-Jacobi
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  2. Okhil K. Nag
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  3. Vibudhuta Awasthi
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  4. Mark A. Babizhayev
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  5. H. Anne Pereira
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Corresponding author

Correspondence to Anne Kasus-Jacobi .

Editor information

Editors and Affiliations

  1. Innovative Vision Products Inc., Moscow, Russia, Innovative Vision Products, Inc.,, County of New Castle, Delaware, USA

    Mark A. Babizhayev

  2. Department of Ophthalmology and Visual Sciences, Truhlsen Eye Institute, College of Medicine, University of Nebraska Medical Center; Key Laboratory of Protein Chemistry and Developmental Biology of Education Ministry of China, College of Life Sciences, Hunan Normal University, Changsha, Hunan, China, Omaha, Nebraska, USA

    David Wan-Cheng Li

  3. Department of Pharmaceutical Sciences and Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA

    Anne Kasus-Jacobi

  4. Department of Ophthalmology, University of Pristina, Faculty of Medicine, Kosovska Mitrovica, Serbia

    Lepša Žorić

  5. Ophthalmology, Miguel Hernandez University, Alicante, Spain

    Jorge L. Alió

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Kasus-Jacobi, A., Nag, O.K., Awasthi, V., Babizhayev, M.A., Pereira, H.A. (2015). Investigating Carcinine Transport and the Expression Profile of Transporter Genes in Human Corneal Epithelial Cells. In: Babizhayev, M., Li, DC., Kasus-Jacobi, A., Žorić, L., Alió, J. (eds) Studies on the Cornea and Lens. Oxidative Stress in Applied Basic Research and Clinical Practice. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1935-2_8

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