Claus Cursiefen / Felix Bock / Deniz Hos - B 3

Novel therapeutic role of myeloid cell-derived VEGF-C and lymphangiogenesis in resolution of corneal inflammation and reestablishment of corneal transparency

Corneal lymphangiogenesis and activated myeloid cells were so far mainly studied in the context of the induction of diseases such as dry eye or graft rejection (1-5). In this project we want to test the novel therapeutic concept of using “immunomodulatory” myeloid cells and isolated lymphangiogenesis to resolve inflammation and regain corneal transparency. In this context we further want to better understand the role of IL10 as well as VEGF-C/D in the induction of immunomodulatory and lymphangiogenic corneal myeloid cell subsets under normal and inflammatory conditions in relation to lymphatics. 

Introduction

In the past decade we and others could show that the presence of myeloid cells, especially macrophages and activated DCs, and the presence of pathologic lymphatic vessels in the cornea significantly increase the risk of corneal graft rejection and developed strategies to reduce the lymphatic load pre- and post-transplantation aiming to achieve a better graft outcome. These strategies have already entered translational phase II and III clinical trials.

Currently, it is unclear whether corneal lymphatic vessels also have physiological functions, e.g. facilitating wound healing after tissue damage. Furthermore, it is not even known whether selective ingrowths of lymphatic vessels occurs  after corneal injury and whether lymphangiogenesis plays a role in the regulation of corneal edema or the drainage of cell debris. Also are the mechanisms whereby this putative anti-inflammatory lymphangiogenesis would be mediated and the immune cells involved unknown (1-5).

IL-10 indirectly regulates corneal lymphangiogenesis and resolution of inflammation via macrophages

Whereas the majority of corneal immunology research was directed towards blocking pathologic lymphangiogenesis (LA) in the context of transplantation, we very recently could show an unexpected beneficial effect of IL-10 polarized macrophages upregulating VEGF-C on the resolution of corneal inflammation (6). Thereby we could for the first time demonstrate a “beneficial” effect of corneal LA. This strongly suggests the hypothesis of a novel therapeutically applicable role of isolated corneal LA driven by immune modulatory myeloid cells in the resolution of blinding corneal transparency loss caused by inflammation and injury.

AIMS: Novel therapeutic role of myeloid cell-derived VEGF-C and lymphangiogenesis in resolution of corneal inflammation and reestablishment of corneal transparency

In the current project we first want to better understand the role of anti-inflammatory cytokines such as IL-10 as well as lymphangiogenic growth factors such as VEGF-C/-D on the immunomodulatory potential of corneal myeloid cell subsets under normal and inflammatory conditions in relation to lymphatics. 

Secondly we want to test the novel therapeutic concept of using these “immunomodulatory” myeloid cells and isolated lymphangiogenesis (LA) to resolve inflammation and regain corneal transparency in vivo in murine models of chemical burn, inflammatory hem- and lymphangiogenesis as well as perforating injury. 

AIMS: Corneal lymphangiogenesis is involved in the regulation of corneal edema and transparency

To date, a physiological role for lymphangiogenesis (LA) in the cornea has not been described. Using a mouse model of perforating corneal injury that causes acute and severe fluid accumulation in the cornea, we have recently shown that lymphatics transiently and selectively invade the cornea and regulate the resolution of corneal edema (7). Furthermore, pharmacological blockade of LA via VEGFR-3 inhibition resulted in increased corneal thickness due to delayed drainage of corneal edema and a trend towards prolonged corneal opacification. Notably, lymphatics were also detectable in the cornea of a patient with acute edema due to spontaneous Descemet´s (basement) membrane rupture in keratoconus, mimicking this animal model and highlighting the clinical relevance of LAs in corneal fluid homeostasis. Together, this work provides first evidence that LA plays an unexpectedly beneficial role in the regulation of corneal edema and transparency. This might open new treatment options in blinding diseases associated with corneal edema and transparency loss. 

Perspectives 

Translationally, we hope to find new approaches to solve persisting corneal inflammation and support wound healing as well as to establish new strategies to regain corneal transparency. These new therapies have not only relevance in Ophthalmology but might also be adaptable to diseases outside the eye like impaired diabetic skin wound healing, psoriasis, or cutaneous chemical and physical burns.

Selected publications

Cursiefen C, Maruyama K, Bock F, Saban D, Sadrai Z, Lawler J, Dana R, Masli S. Thrombospondin 1 inhibits inflammatory lymphangiogenesis by CD36 ligation on monocytes. J Exp Med. (2011);208:1083-92. (IF=13)

Bock F, Maruyama K, Regenfuss B, Hos D, Steven P, Heindl LM, Cursiefen C. Novel anti(lymph)angiogenic treatment strategies for corneal and ocular surface diseases. Prog Retin Eye Res. (2013);34:89-124. (IF=12) 

Platonova N, Miquel G, Regenfuss B, Taouji S, Cursiefen C, Chevet E, Bikfalvi A. Evidence for the interaction of fibroblast growth factor-2 with the lymphatic endothelial cell marker LYVE-1. Blood. (2013);121:1229-37. (IF=13)

Cursiefen C, Viaud E, Bock F, et al.  Aganirsen antisense oligonucleotide eye drops inhibit keratitis-induced corneal neovascularization and reduce need for transplantation: The I-CAN study. Ophthalmology. (2014);121:1683-92. (IF=8)

Bock F, Matthaei M, Reinhard T, Böhringer D, Christoph J, Ganslandt T, Cursiefen C. High-dose subconjunctival cyclosporine a implants do not affect corneal neovascularization after high-risk keratoplasty. Ophthalmology. (2014);121:1677-82. (IF=8)

Hos D, Bucher F, Regenfuss B, Dreisow ML, Bock F, Heindl LM, Eming SA, Cursiefen C. IL-10 indirectly regulates corneal lymphangiogenesis and resolution of inflammation via macrophages. Am J Pathol. (2016);186:159-71. (IF=5)

Hos D, Bukowiecki A, Horstmann J, Bock F, Bucher F, Heindl LM, Siebelmann S, Steven P, Dana R, Eming SA, Cursiefen C. Transient Ingrowth of Lymphatic Vessels into the Physiologically Avascular Cornea Regulates Corneal Edema and Transparency. Sci Rep. 2017 Aug 3;7(1):7227. (IF=5)

Hos D, Tuac O, Schaub F, Stanzel TP, Schrittenlocher S, Hellmich M, Bachmann BO, Cursiefen C. Incidence and Clinical Course of Immune Reactions after Descemet Membrane Endothelial Keratoplasty: Retrospective Analysis of 1000 Consecutive Eyes. Ophthalmology. 2017 Apr;124(4):512-518 (IF=8)


Prof. Dr. Claus Cursiefen

Dept. of Ophthalmology / RG location - LFI Building

Prof. Dr. Claus Cursiefen

Principal Investigator B 3
Executive Board Member

claus.cursiefen@uk-koeln.de

Work +49 221 478 4300

Eye Center Cologne
Kerpener Str. 62
50937 Cologne

https://augenklinik.uk-koeln.de/zentrum/direktoren-und-teams/

Publications - Claus Cursiefen

Link to PubMed


Dr. Felix Bock

Dept. of Ophthalmology / RG location - LFI Building

Dr. Felix Bock

Co-Principal Investigator B 3

felix.bock@uk-koeln.de

Work +49 221 478 98896

Building 13
Kerpener Str. 62
50924 Cologne

Publications - Felix Bock

Link to PubMed


Dr. Dr. Deniz Hos

Dept. of Ophthalmology / RG location - LFI Building

Dr. Dr. Deniz Hos

Co-Principal Investigator B 3 / Principal Investigator CAP 11

deniz.hos@uk-koeln.de

Work +49 221 478 98896

Building 13
Kerpener Str. 62
50924 Cologne

Publications - Deniz Hos

Link to PubMed

Group Members

Maria Nortara (Postdoc)
Thomas Clahsen (Postdoc)
Ann-Charlott Schneider (doctoral student)
Anne Bukowieki (doctoral student)
Yanhong Hou (doctoral student)
Hung Le (doctoral student)
Gabriele Braun (technician)
Sara Beboudifard (technician)

Figure 1

CMMC Cursiefen
Corneal cross sections after injury show the occurrence of Il-10 positive (prolymphangiogenic) macrophages

Figure 2

Perforating corneal injury induces transient edema and isolated lymph- without hemangiogenesis. (A) Model of perforating corneal injury. (B–E) In vivo optical coherence tomography scans demonstrate a transient increase of corneal thickness, green values: central corneal thickness. (F–I) Corneal whole mounts stained with LYVE-1 (red) and CD31 (green); LYVE-1high/CD31low lymphatic vessels (arrows) growing towards the central cornea are detectable 1 week after injury and persist until 2 weeks after injury. Afterwards, lymphatic vessels regress and are comparable to uninjured corneas after 4 weeks. In contrast to lymphatic vessels, no significant ingrowth of LYVE-1neg/CD31high blood vessels is detectable; p.i.: post injury (8).