July 2019
Volume 60, Issue 9
Free
ARVO Annual Meeting Abstract  |   July 2019
Indian hedgehog (Ihh) secreted by adult choroid endothelial cells` regulates choroidal homeostasis and immune response
Author Affiliations & Notes
  • Christin Hanke-Gogokhia
    Ophthalmology, Weill Cornell Medical College, New York, United States
  • Guillermo Lehmann
    Ophthalmology, Weill Cornell Medical College, New York, United States
    Regeneron Pharmaceuticals, New York, United States
  • Yang Hu
    Physiology and Biophysics, Weill Cornell Medical College, New York, United States
  • Rohan Bareja
    Physiology and Biophysics, Weill Cornell Medical College, New York, United States
  • Micheal Ginsberg
    Angiocrine Bioscience, San Diego, California, United States
  • Daniel Nolan
    Angiocrine Bioscience, San Diego, California, United States
  • Alexandre Wojcinski
    Memorial Sloan Kettering Cancer Center, New York, United States
  • Robert F Mullins
    Institute for Vision Research, University of Iowa, Iowa, United States
  • Gerard A Lutty
    Wilmer Ophthalmological Institute, Johns Hopkins Hospital, Maryland, United States
  • Olivier Elemento
    Physiology and Biophysics, Weill Cornell Medical College, New York, United States
  • Alexandra Joyner
    Memorial Sloan Kettering Cancer Center, New York, United States
  • Shahin Rafii
    Regenerative Medicine, Weill Cornell Medical College, New York, United States
  • Ignacio Benedicto
    Ophthalmology, Weill Cornell Medical College, New York, United States
    Centro Nacional de Investigaciones Cardiovasculares, Madrid, Spain
  • Enrique J Rodriguez-Boulan
    Ophthalmology, Weill Cornell Medical College, New York, United States
  • Footnotes
    Commercial Relationships   Christin Hanke-Gogokhia, None; Guillermo Lehmann, None; Yang Hu, None; Rohan Bareja, None; Micheal Ginsberg, None; Daniel Nolan, None; Alexandre Wojcinski, None; Robert Mullins, None; Gerard Lutty, None; Olivier Elemento, None; Alexandra Joyner, None; Shahin Rafii, None; Ignacio Benedicto, None; Enrique Rodriguez-Boulan, None
  • Footnotes
    Support  NIH (R01 EY08531), Starr Foundation, New York State Stem Cell Program, Research to Prevent Blindness, Dyson Foundation to the Ophthalmology Department
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 5238. doi:
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      Christin Hanke-Gogokhia, Guillermo Lehmann, Yang Hu, Rohan Bareja, Micheal Ginsberg, Daniel Nolan, Alexandre Wojcinski, Robert F Mullins, Gerard A Lutty, Olivier Elemento, Alexandra Joyner, Shahin Rafii, Ignacio Benedicto, Enrique J Rodriguez-Boulan; Indian hedgehog (Ihh) secreted by adult choroid endothelial cells` regulates choroidal homeostasis and immune response. Invest. Ophthalmol. Vis. Sci. 2019;60(9):5238.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Patients with atrophic AMD display photoreceptor loss secondary to RPE dysfunction and choroidal damage. However, the exact etiology of AMD remains unknown due to lack of information on RPE/choroid cell diversity and intercellular crosstalk mechanisms. Here we report the selective high expression of Indian Hedgehog (Ihh) in adult choroid endothelial cells (ECs) and its function as an angio- and immunomodulatory choroid signaling molecule likely relevant to AMD.

Methods : Cells from tissue of wildtype RPE/choroid were single-cell sorted and scRNAseq was performed. Bulk RNAseq was used to compare the transcriptomes of purified RPE/choroid from wildtype and mutant mice after laser-induced CNV. To study crosstalk between EC-derived Ihh and stromal, Gli1+ perivascular cells, adult tamoxifen-induced knockout mice were generated by crossing Ihhflox/flox with Cdh5-Ert2Cre+ transgenic mice, and cDNA of eGFP was inserted into exon 1 of mouse Gli1 gene to generate Gli1+/eGFP knock-in animals. Photoreceptor function was measured by ERG and OKT, and morphology of RPE/choroid and photoreceptors was evaluated by immunohistochemistry. In-vitro assays such as flow-cytometry and RT-qPCR were used to examine crosstalk between RPE and choroid.

Results : We report the first single cell RNAseq analysis of adult mouse RPE/choroid tissue identifying 13 main cell types including 3 subtypes of ECs. By combining these results with a transcriptomic analysis of tissue-specific ECs from other organs, we found a marked enrichment of Ihh expression in choroid ECs, particularly in the choriocapillaries located underneath the RPE. Using reporter mice, we identified the target of choroidal Ihh signaling as a large population of stromal, Gli1+ perivascular mesenchymal stem cell (MSC)-like cells. EC-specific Ihh KO and Gli1eGFP/eGFP mice displayed loss of mast cells, altered inflammatory and vasculatory response, visual impairment and retinal degeneration indicating a novel Ihh/Gli1-dependent signaling circuit in adult choroid regulating choroidal homeostasis and retinal function.

Conclusions : Our study uncovers the cellular and molecular landscape of the adult mouse RPE/choroid and stromal Gli1+ MSC-like cells as key player in response to EC-derived Ihh implicated in choroidal neovascularization and immune response, thus providing possible new therapeutic targets for atrophic AMD and choroidal disorders.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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