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Angiopoietin receptor TEK interacts with CYP1B1 in primary congenital glaucoma

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Abstract

Primary congenital glaucoma (PCG) is a severe autosomal recessive ocular disorder associated with considerable clinical and genetic heterogeneity. Recently, rare heterozygous alleles in the angiopoietin receptor-encoding gene TEK were implicated in PCG. We undertook this study to ascertain the second mutant allele in a large cohort (n = 337) of autosomal recessive PCG cases that carried heterozygous TEK mutations. Our investigations revealed 12 rare heterozygous missense mutations in TEK by targeted sequencing. Interestingly, four of these TEK mutations (p.E103D, p.I148T, p.Q214P, and p.G743A) co-occurred with three heterozygous mutations in another major PCG gene CYP1B1 (p.A115P, p.E229K, and p.R368H) in five families. The parents of these probands harbored either of the heterozygous TEK or CYP1B1 alleles and were asymptomatic, indicating a potential digenic mode of inheritance. Furthermore, we ascertained the interactions of TEK and CYP1B1 by co-transfection and pull-down assays in HEK293 cells. Ligand responsiveness of the wild-type and mutant TEK proteins was assessed in HUVECs using immunofluorescence analysis. We observed that recombinant TEK and CYP1B1 proteins interact with each other, while the disease-associated allelic combinations of TEK (p.E103D)::CYP1B1 (p.A115P), TEK (p.Q214P)::CYP1B1 (p.E229K), and TEK (p.I148T)::CYP1B1 (p.R368H) exhibit perturbed interaction. The mutations also diminished the ability of TEK to respond to ligand stimulation, indicating perturbed TEK signaling. Overall, our data suggest that interaction of TEK and CYP1B1 contributes to PCG pathogenesis and argue that TEK-CYP1B1 may perform overlapping as well as distinct functions in manifesting the disease etiology.

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Acknowledgements

We thank the PCG, POAG, and anterior segment dysgenesis patients and their families, and the normal volunteers for their participation in this study. The authors acknowledge the help of Drs. Ravi Thomas, Rajul Parikh, Harsha L Rao, and Garudadri C Sekhar for the diagnosis of some patients and controls; Drs. Aramati BM Reddy, Kiranpreet Kaur, Ratnakar Tripathi, and Mr. Chandan S Appikonda for sample collection; and Dr. Prathap Naidu and Mr. Hersh Parikh for the technical help. The authors would like to thank the Exome Aggregation Consortium and the groups that provided exome variant data for comparison. A full list of contributing groups can be found at http://exac.broadinstitute.org/about.

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Author notes

  1. Meha Kabra and Wei Zhang were equally contributing authors.

Authors and Affiliations

  1. Kallam Anji Reddy Molecular Genetics Laboratory, Brien Holden Eye Research Centre, L.V. Prasad Eye Institute, Hyderabad, India

    Meha Kabra, Sonika Rathi, Goutham Pyatla, Inderjeet Kaur & Subhabrata Chakrabarti

  2. Jasti V Ramanamma Children’s Eye Care Centre, L.V. Prasad Eye Institute, Hyderabad, India

    Anil K. Mandal & Sirisha Senthil

  3. VST Centre for Glaucoma Care, L.V. Prasad Eye Institute, Hyderabad, India

    Anil K. Mandal & Sirisha Senthil

  4. Tej Kohli Cornea Institute, L.V. Prasad Eye Institute, Hyderabad, India

    Muralidhar Ramappa

  5. Gullapalli Pratibha Rao International Centre for Advancement of Rural Eye Care, L.V. Prasad Eye Institute, Hyderabad, India

    Seema Banerjee, Konegari Shekhar, Srinivas Marmamula, Asha L. Mettla & Rohit C. Khanna

  6. Department of Ophthalmology, UMASS Medical School, Worcester, MA, USA

    Wei Zhang & Hemant Khanna

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  1. Meha Kabra
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Correspondence to Subhabrata Chakrabarti.

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Funding

This work was supported by grants from the Department of Biotechnology, Government of India (BT/01/COE/06/02/10) to SC, and the National Eye Institute, NIH (EY022372) to HK. GP was supported by a fellowship from the Council of Scientific and Industrial Research, Government of India.

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The authors declare that they have no conflict of interest.

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All the molecular genetic, biochemical, and cellular data generated during this study are included in this manuscript and its supplementary files. However, the patients’ clinical data are confidential and will be available from the corresponding author on request.

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Kabra, M., Zhang, W., Rathi, S. et al. Angiopoietin receptor TEK interacts with CYP1B1 in primary congenital glaucoma. Hum Genet 136, 941–949 (2017). https://doi.org/10.1007/s00439-017-1823-6

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