Center for Molecular Medicine Cologne

Notara, Maria | Cursiefen, Claus - A 09

Role and therapeutic potential of autophagy-related mechanisms in UV-induced blinding ocular stem cell disorders (Pterygium)

Introduction

UV-induced LESC damage causing pterygium is prevalent in sunny and dry climates, but also affects patients in Northern Europe (1-10%). It is estimated that an expected 10% of all refugees who came to Germany from southern countries are suffering from pterygium (i.e.>100.000), making new treatments more imperative. Understanding the mechanisms by which autophagy changes contribute to pterygium pathogenesis will lead to new therapies to limit recurrence, a major problem after pterygium surgery.

Vision is a key sensory function. The transparent and avascular cornea (“windscreen of the eye”) is essential for sight. It’s primary refractive layer, the corneal epithelium, is maintained via ABCB5-expressing limbal epithelial stem cells (LESC) located at the junction between the avascular cornea and the densely (lymph)vascularized conjunctiva.
Due to their position, LESC are exposed to UV irradiation, which can damage their niche and function. Increased levels of autophagic activity within the basal limbal epithelium indicate that LESCs employ autophagy to repair damage accumulating during their long cycle. Pterygium, a tumour of the conjunctiva which grows onto the cornea impairing vision is linked to UV-induced LESC damage. We have previously reported that LESCs differentiate under UV exposure, leading to a pro-inflammatory shift in their niche. Pterygium is linked to reduced autophagy via mTORC1in the epithelium which boosts proliferation and causes aberrant apoptosis. The direct involvement of the LESCS in pterygium pathogenesis via this mechanism is unclear.
Therefore, we aim to:

  1. elucidate the molecular mechanisms by which autophagy affects the maintenance of ABCB5-positive LESCs phenotype and how these change in pterygium and
  2. unravel if and how UV light damages ABCB5-positive LESCs via reduction of autophagic activity.

A better understanding of the autophagic activity mechanisms exerted by LESC and their damage by UV light will open the way to new treatment options in the form of mTOR inhibitors against UV-related stem cell disorders in the eye and beyond.

Our Aims

1. Assessment of the role of autophagy in UVB-response in pterygium, limbus and conjunctiva.

  • In situ assessment of autophagy –related proteins and putative stem cell marker distribution in primary and recurrent pterygium, limbus and conjunctiva.
  • Assessment of autophagy induction in the limbal stem cell compartment following UVB irradiation by UVB irradiation of human pterygium, limbal and conjunctiva ABCB5-positive stem cells.
  • Evaluation of the function of autophagy following UVB irradiation of human pterygium, limbal and conjunctiva.
  • Paracrine effect of autophagy deficient-sufficient UVB treated/non-treated ABCB5 positive and negative cells from pterygium, limbus and conjunctiva 2. Use of in vivo models to evaluate the role of autophagy following UVB irradiation (collaboration with Prof. Björn Schumacher)

2. Assessment of the autophagy response to UV-irradiation of an ABCB5-KO model

  • We will study the response of the autophagy-defficent mice to UVB treatment in terms of assessment of lymphangiogenesis as well as assessment of immune cell infiltration

Previous Work

Our group has recently used an in vitro approach to study the effect of UVA and UVB on the phenotype and functionality of human limbal epithelial cells and their accessory limbal fibroblasts, as well as on their paracrine signalling activities associated with the regulation of inflammation and (lymph)angiogenesis1,2. Our findings suggested that this short-term UV irradiation induced the loss of the stem-like character of limbal epithelial cells (Figure 1). Interestingly, limbal epithelial cells co-cultured with UV-irradiated limbal fibroblasts also exhibited loss of SC character and decrease of colony-forming efficiency1,2.

Figure 1

Following UVB treatment, pro-angiogenic factors were down regulated while pro-inflammatory and macrophage-recruiting cytokines such as TNF-α, IFNγ and MCP1 were significantly upregulated (Figure 2).

Figure 2

This demonstrates that by producing these cytokines, both HLE and HLF cells may contribute to the inflammatory mechanisms taking place in the cornea following UVB irradiation. In addition, we have recently demonstrated that UV blocking contact prevented these UVB-induced adverse effects as well as DNA damage. These data demonstrated the significance of UV-protection in preserving the limbal niche in response to UVB irradiation, suggesting that protecting the niche, especially after LESC transplantation and in patients after pterygium surgery, may prevent recurrences3.Taken together, these observations demonstrate that UV irradiation dysregulates limbal niche cells while inducing a pro-inflammatory environment which promotes neovascularisation.
Although UV-induced changes in the limbal niche have already been described, the exact mechanisms through which these occur remain unidentified. Recent preliminary studies have shown that autophagy regulates cell cycle response in limbal epithelial cultures under UVA stress and that autophagy-deficient cells are not able to restore reactive oxygen species accumulation mediated by the nuclear to cytoplasm translocation of pax6; however, the events following UVB treatment as well as the specific effects on the stem cell population have not been investigated. This project will aim to address these questions.

  • Notara, M.,… and Cursiefen, Stem Cell Res, 2015. 15(3): p. 643-54.
  • Notara, M., … and Cursiefen, Invest Ophthalmol Vis Sci, 2016. 57(3): p. 928-39.
  • Notara, M.,… and Cursiefen, Sci Rep, 2018. 8(1): p. 12564.
  • Notara, M … and Cursiefen, Journal of Clinical Medicine, 2019 Nov 9;8(11)
  • Notara, M … and Cursiefen, Stem Cells International., 2018 May 8;2018:8620172.
  • Hos, D, …, Notara,… and Cursiefen, Prog Retin Eye Res. 2019 Nov;73:100768.
  • Ozer O, Mestanoglu M, Howaldt A, Clahsen T, Schiller P, Siebelmann S, Reinking N, Cursiefen C, Bachmann B, and Matthaei M (2022). Correlation of Clinical Fibrillar Layer Detection and Corneal Thickness in Advanced Fuchs Endothelial Corneal Dystrophy. J Clin Med11. doi:10.3390/jcm11102815.
  • Volatier T, Schumacher B, Cursiefen C, Notara M. UV Protection in the Cornea:Failure and Rescue. Biology (Basel). 2022 Feb 10;11(2):278. doi:10.3390/biology11020278.
  • Zhang W, Schonberg A, Bock F, and Cursiefen C (2022). Posttransplant VEGFR1R2 Trap Eye Drops Inhibit Corneal (Lymph)angiogenesis and Improve Corneal Allograft Survival in Eyes at High Risk of Rejection. Transl Vis Sci Technol11, 6. doi:10.1167/tvst.11.5.6.
  • Peil J, Bock F, Kiefer F, Schmidt R, Heindl LM, Cursiefen C, and Schlereth SL (2022). New Therapeutic Approaches for Conjunctival Melanoma-What We Know So Far and Where Therapy Is Potentially Heading: Focus on Lymphatic Vessels and Dendritic Cells. Int J Mol Sci23. doi:10.3390/ijms23031478.
  • Schrittenlocher S, Grass C, Dietlein T, Lappas A, Matthaei M, Cursiefen C, and Bachmann B (2022). Graft survival of Descemet membrane endothelial keratoplasty (DMEK) in corneal endothelial decompensation after glaucoma surgery. Graefes Arch Clin Exp Ophthalmol260, 1573-1582. doi:10.1007/s00417-021-05506-4.
  • Schrittenlocher S, Matthaei M, Bachmann B, and Cursiefen C (2022). The Cologne-Mecklenburg-Vorpommern DMEK Donor Study (COMEDOS) - design and review of the influence of donor characteristics on Descemet membrane endothelial keratoplasty (DMEK) outcome. Graefes Arch Clin Exp Ophthalmol. doi:10.1007/s00417-022-05594-w.
  • Volatier T, Schumacher B, Cursiefen C, and Notara M (2022). UV Protection in the Cornea: Failure and Rescue. Biology (Basel)11. doi:10.3390/biology11020278.
  • Zwingelberg SB, Buscher F, Schrittenlocher S, Rokohl AC, Loreck N, Wawer-Matos P, Fassin A, Schaub F, Roters S, Matthaei M, Heindl LM, Bachmann BO, and Cursiefen C (2022). Long-Term Outcome of Descemet Membrane Endothelial Keratoplasty in Eyes With Fuchs Endothelial Corneal Dystrophy Versus Pseudophakic Bullous Keratopathy. Cornea41, 304-309. doi:10.1097/ICO.0000000000002737.
  • Handel A, Luke JN, Siebelmann S, Franklin J, Roters S, Matthaei M, Bachmann BO, Cursiefen C, and Hos D (2022). Outcomes of deep anterior lamellar keratoplasty and penetrating keratoplasty in keratoconic eyes with and without previous hydrops. Graefes Arch Clin Exp Ophthalmol. doi:10.1007/s00417-022-05643-4.
  • Handel A, Siebelmann S, Luke JN, Matthaei M, Cursiefen C, and Bachmann B (2022). Influence of Body Position on Intraocular Pressure After Descemet Membrane Endothelial Keratoplasty: A Prospective Randomized Trial. Cornea. doi:10.1097/ICO.0000000000003010.
  • Hribek A, Mestanoglu M, Clahsen T, Reinking N, Frentzen F, Howaldt A, Siebelmann S, Bachmann BO, Cursiefen C, and Matthaei M (2022). Scheimpflug Backscatter Imaging of the Fibrillar Layer in Fuchs Endothelial Corneal Dystrophy. Am J Ophthalmol235, 63-70. doi:10.1016/j.ajo.2021.08.019.
  • Mestanoglu M, Handel A, Cursiefen C, and Hos D (2022). Three-year follow-up of high-risk keratoplasty following fine-needle diathermy of corneal neovascularization combined with bevacizumab. Graefes Arch Clin Exp Ophthalmol 260, 2383-2385. doi:10.1007/s00417-021-05546-w.
  • Roters S, Rokohl AC, Heindl LM, and Cursiefen C (2022). Novel eccentric corneoscleral donor preparation technique providing corneoscleral tectonic and central split corneal grafts for multiple recipients. Graefes Arch Clin Exp Ophthalmol 260, 2069-2071. doi:10.1007/s00417-021-05482-9.
  • Schaub F, Bachmann BO, and Cursiefen C (2022). Silicone oil endotamponade in eyes with Boston Keratoprosthesis Type 1. Acta Ophthalmol 100, e1041-e1042. doi:10.1111/aos.15026.
  • Schaub F, Mestanoglu M, Cursiefen C, and Hos D (2022). Impact of early intensified postoperative corticosteroids on immune reaction rates after Descemet membrane endothelial keratoplasty (DMEK). Graefes Arch Clin Exp Ophthalmol 260, 693-695. doi:10.1007/s00417-021-05393-9.
  • Hou Y, Bock F, Hos D, and Cursiefen C (2021). Lymphatic Trafficking in the Eye: Modulation of Lymphatic Trafficking to Promote Corneal Transplant Survival. Cells10. doi:10.3390/cells10071661.
  • Norrick A, Esterlechner J, Niebergall-Roth E, Dehio U, Sadeghi S, Schroder HM, Ballikaya S, Stemler N, Ganss C, Dieter K, Dachtler AK, Merz P, Sel S, Chodosh J, Cursiefen C, Frank NY, Auffarth GU, Ksander B, Frank MH, and Kluth MA (2021). Process development and safety evaluation of ABCB5(+) limbal stem cells as advanced-therapy medicinal product to treat limbal stem cell deficiency. Stem Cell Res Ther12, 194. doi:10.1186/s13287-021-02272-2.
  • Schlereth SL, Hos D, Matthaei M, Hamrah P, Schmetterer L, O'Leary O, Ullmer C, Horstmann J, Bock F, Wacker K, Schroder H, Notara M, Haagdorens M, Nuijts R, Dunker SL, Dickman MM, Fauser S, Scholl HPN, Wheeler-Schilling T, and Cursiefen C (2021). New Technologies in Clinical Trials in Corneal Diseases and Limbal Stem Cell Deficiency: Review from the European Vision Institute Special Interest Focus Group Meeting. Ophthalmic Res64, 145-167. doi:10.1159/000509954.
  • Handel A, Siebelmann S, Hos D, Ogrunc F, Matthaei M, Cursiefen C, and Bachmann B (2021). Comparison of Mini-DMEK versus predescemetal sutures as treatment of acute hydrops in keratoconus. Acta Ophthalmol99, e1326-e1333. doi:10.1111/aos.14835.
  • Hayashi T, Zhang W, Hos D, Schrittenlocher S, Nhat Hung Le V, Siebelmann S, Matthaei M, Bock F, Bachmann B, and Cursiefen C (2021). Descemet Membrane Endothelial Keratoplasty in Vascularized Eyes: Outcome and Effect on Corneal Neovascularization. Cornea40, 685-689. doi:10.1097/ICO.0000000000002502.
  • Hribek A, Mestanoglu M, Clahsen T, Reinking N, Frentzen F, Howaldt A, Siebelmann S, Bachmann BO, Cursiefen C, and Matthaei M (2021). Scheimpflug Backscatter Imaging of the Fibrillar Layer in Fuchs Endothelial Corneal Dystrophy. Am J Ophthalmol235, 63-70. doi:10.1016/j.ajo.2021.08.019.
  • Schrittenlocher S, Grass C, Dietlein T, Lappas A, Matthaei M, Cursiefen C, and Bachmann B (2021). Graft survival of Descemet membrane endothelial keratoplasty (DMEK) in corneal endothelial decompensation after glaucoma surgery. Graefes Arch Clin Exp Ophthalmol. doi:10.1007/s00417-021-05506-4.
  • Zhang W, Schonberg A, Hamdorf M, Georgiev T, Cursiefen C, and Bock F (2021). Preincubation of donor tissue with a VEGF cytokine trap promotes subsequent high-risk corneal transplant survival. Br J Ophthalmol. doi:10.1136/bjophthalmol-2021-319745.
  • Cursiefen C, and Hos D (2021). Cutting Edge: Novel Treatment Options Targeting Corneal Neovascularization to Improve High-Risk Corneal Graft Survival. Cornea40, 1512-1518. doi:10.1097/ICO.0000000000002736.
  • Hadrian K, Willenborg S, Bock F, Cursiefen C, Eming SA, and Hos D (2021). Macrophage-Mediated Tissue Vascularization: Similarities and Differences Between Cornea and Skin. Front Immunol12, 667830. doi:10.3389/fimmu.2021.667830.
  • Hos, D., Matthaei, M., Bock, F., Maruyama, K., Notara, M., Clahsen, T., Hou, Y., Le, V.N.H., Salabarria, A.C., Horstmann, J., Bachmann, B.O., and Cursiefen, C. (2019). Immune reactions after modern lamellar (DALK, DSAEK, DMEK) versus conventional penetrating corneal transplantation. Prog Retin Eye Res10.1016/j.preteyeres.2019.07.001.
  • Notara M, Lentzsch A, Clahsen T, Behboudifard S, Braun G, Cursiefen C: Bevacizumab Induces Upregulation of Keratin 3 and VEGFA in Human Limbal Epithelial Cells in Vitro. J Clin Med. 2019 Nov 9;8(11):1925. doi: 10.3390/jcm8111925. PMID: 31717500
  • Clahsen T, Buttner C, Hatami N, Reis A, and Cursiefen C (2020). Role of Endogenous Regulators of Hem- And Lymphangiogenesis in Corneal Transplantation. J Clin Med 9.
  • Hayashi T, Schrittenlocher S, Siebelmann S, Le VNH, Matthaei M, Franklin J, Bachmann B, and Cursiefen C (2020a). Risk factors for endothelial cell loss after Descemet membrane endothelial keratoplasty (DMEK). Sci Rep-Uk 10.
  • Le VNH, Hos D, Hou Y, Witt M, Barkovskiy M, Bock F, and Cursiefen C (2020a). VEGF TrapR1R2 Suspended in the Semifluorinated Alkane F6H8 Inhibits Inflammatory Corneal Hem- and Lymphangiogenesis. Translational vision science & technology 9, 15.
  • Le VNH, Hou Y, Bock F, and Cursiefen C (2020b). Supplemental Anti Vegf A-Therapy Prevents Rebound Neovascularisation After Fine Needle Diathermy Treatment to Regress Pathological Corneal (LYMPH)Angiogenesis. Sci Rep 10, 3908.
  • Salabarria AC, Koch M, Schonberg A, Zinser E, Hos D, Hamdorf M, Imhof T, Braun G, Cursiefen C, and Bock F (2020). Topical VEGF-C/D Inhibition Prevents Lymphatic Vessel Ingrowth into Cornea but Does Not Improve Corneal Graft Survival. J Clin Med 9.
  • Schaub F, Collmer M, Schrittenlocher S, Bachmann BO, Cursiefen C, and Hos D (2020). Outcome of Descemet Membrane Endothelial Keratoplasty Using Corneas from Donors >/=80 Years of Age. Am J Ophthalmol 211, 200-6.
  • Schrittenlocher S, Schlereth SL, Siebelmann S, Hayashi T, Matthaei M, Bachmann B, and Cursiefen C (2020). Long-term outcome of descemet membrane endothelial keratoplasty (DMEK) following failed penetrating keratoplasty (PK). Acta Ophthalmol 10.1111/aos.14417.
  • Siggel R, Schroedl F, Dietlein T, Koch KR, Platzl C, Kaser-Eichberger A, Cursiefen C, and Heindl LM (2020). Absence of lymphatic vessels in non-functioning bleb capsules of glaucoma drainage devices. Histology and histopathology 10.14670/HH-18-300, 18300.

 

Dr. Maria Notara CMMC Cologne
Dr. Maria Notara

Clinic of General Ophthalmology

CMMC - PI - A 09

CMMC - PI - CAP 15

+49 221 478 30621

+49 221 478 32785

Clinic of General Ophthalmology

Kerpener Str. 62

50931 Köln

https://augenklinik.uk-koeln.de/forschung/zentrum-fuer-molekulare-medizin-koeln-zmmk/

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Prof. Dr. Claus Cursiefen CMMC Cologne
Prof. Dr. Claus Cursiefen

Clinic of General Ophthalmology

CMMC - Co-PI - A 09

Executive Board Member

+49 221 478 4300

+49 221 478 97295

Clinic of General Ophthalmology

Kerpener Str. 62

50937 Cologne

https://augenklinik.uk-koeln.de/forschung/arbeitsgruppen-labore/cornea-lab/

CMMC Profile Page

Curriculum Vitae (CV)

Publications on PubMed

Publications - Claus Cursiefen

Link to PubMed