Repurposing of glycine transport inhibitors for the treatment of erythropoietic protoporphyria

François Halloy; Pavithra S. Iyer; Alice Ghidini; Veronika Lysenko; Jasmin Barman-Aksözen; Chia Pei Grubenmann; Jessica Jucker; Nicole Wildner-Verhey van Wijk; Marc David Ruepp; Elisabeth I. Minder; Anna Elisabeth Minder; Xiaoye Schneider-Yin; Alexandre P.A. Theocharides; Daniel Schümperli; Jonathan Hall

doi:10.1016/j.chembiol.2021.02.021

Repurposing of glycine transport inhibitors for the treatment of erythropoietic protoporphyria

François Halloy, Pavithra S. Iyer, Alice Ghidini, Veronika Lysenko, Jasmin Barman-Aksözen, Chia Pei Grubenmann, Jessica Jucker, Nicole Wildner-Verhey van Wijk, Marc David Ruepp, Elisabeth I. Minder, Anna Elisabeth Minder, Xiaoye Schneider-Yin, Alexandre P.A. Theocharides, Daniel Schümperli^*, Jonathan Hall

^*Corresponding author for this work

Basic and Clinical Neuroscience

Research output: Contribution to journal › Article › peer-review

16 Citations (Scopus)

Abstract

Erythropoietic protoporphyria (EPP) is a rare disease in which patients experience severe light sensitivity. It is caused by a deficiency of ferrochelatase (FECH), the last enzyme in heme biosynthesis (HBS). The lack of FECH causes accumulation of its photoreactive substrate protoporphyrin IX (PPIX) in patients' erythrocytes. Here, we explored an approach for the treatment of EPP by decreasing PPIX synthesis using small-molecule inhibitors directed to factors in the HBS pathway. We generated a FECH-knockout clone from K562 erythroleukemia cells, which accumulates PPIX and undergoes oxidative stress upon light exposure. We used these matched cell lines to screen a set of publicly available inhibitors of factors in the HBS pathway. Inhibitors of the glycine transporters GlyT1 and GlyT2 lowered levels of PPIX and markers of oxidative stress selectively in K562^11B4 cells, and in primary erythroid cultures from an EPP patient. Our findings open the door to investigation of glycine transport inhibitors for HBS disorders.

Original language	English
Pages (from-to)	1221-1234.e6
Journal	Cell Chemical Biology
Volume	28
Issue number	8
DOIs	https://doi.org/10.1016/j.chembiol.2021.02.021
Publication status	Published - 19 Aug 2021

Keywords

CRISPR-Cas9
drug repurposing
erythropoietic protoporphyria
glycine transporter 2
oxidative stress
photodamage
protoporphyrin IX

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10.1016/j.chembiol.2021.02.021

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Halloy, F., Iyer, P. S., Ghidini, A., Lysenko, V., Barman-Aksözen, J., Grubenmann, C. P., Jucker, J., Wildner-Verhey van Wijk, N., Ruepp, M. D., Minder, E. I., Minder, A. E., Schneider-Yin, X., Theocharides, A. P. A., Schümperli, D., & Hall, J. (2021). Repurposing of glycine transport inhibitors for the treatment of erythropoietic protoporphyria. Cell Chemical Biology, 28(8), 1221-1234.e6. https://doi.org/10.1016/j.chembiol.2021.02.021

@article{85c2d8f0541c46d9aecf199eb5641519,

title = "Repurposing of glycine transport inhibitors for the treatment of erythropoietic protoporphyria",

abstract = "Erythropoietic protoporphyria (EPP) is a rare disease in which patients experience severe light sensitivity. It is caused by a deficiency of ferrochelatase (FECH), the last enzyme in heme biosynthesis (HBS). The lack of FECH causes accumulation of its photoreactive substrate protoporphyrin IX (PPIX) in patients' erythrocytes. Here, we explored an approach for the treatment of EPP by decreasing PPIX synthesis using small-molecule inhibitors directed to factors in the HBS pathway. We generated a FECH-knockout clone from K562 erythroleukemia cells, which accumulates PPIX and undergoes oxidative stress upon light exposure. We used these matched cell lines to screen a set of publicly available inhibitors of factors in the HBS pathway. Inhibitors of the glycine transporters GlyT1 and GlyT2 lowered levels of PPIX and markers of oxidative stress selectively in K56211B4 cells, and in primary erythroid cultures from an EPP patient. Our findings open the door to investigation of glycine transport inhibitors for HBS disorders.",

keywords = "CRISPR-Cas9, drug repurposing, erythropoietic protoporphyria, glycine transporter 2, oxidative stress, photodamage, protoporphyrin IX",

author = "Fran{\c c}ois Halloy and Iyer, {Pavithra S.} and Alice Ghidini and Veronika Lysenko and Jasmin Barman-Aks{\"o}zen and Grubenmann, {Chia Pei} and Jessica Jucker and {Wildner-Verhey van Wijk}, Nicole and Ruepp, {Marc David} and Minder, {Elisabeth I.} and Minder, {Anna Elisabeth} and Xiaoye Schneider-Yin and Theocharides, {Alexandre P.A.} and Daniel Sch{\"u}mperli and Jonathan Hall",

note = "Funding Information: We thank Dr. Malgorzata Kisielow (ETH Zurich, Flow Cytometry Core Facility) for useful input and discussions on PPIX quantification by cytofluorometry. We thank Andr{\'e} Imboden (ETH Zurich, Department of Biology) for technical lamp information regarding the light photosensitivity assay. We thank Asuka Fry and Patrizia Belleda (University Hospital Zurich) for CD34 + cell isolation. We thank Sylvain Egloff for performing preliminary CRISPR experiments and J. Philipp Becker for assistance with cell culture. This research was supported in part by NCCR RNA & Disease , funded by the Swiss National Science Foundation , and by ETH Z{\"u}rich . A.P.A.T. is supported by the Professor Dr. Max Clo{\"e}tta Foundation . Funding Information: We thank Dr. Malgorzata Kisielow (ETH Zurich, Flow Cytometry Core Facility) for useful input and discussions on PPIX quantification by cytofluorometry. We thank Andr? Imboden (ETH Zurich, Department of Biology) for technical lamp information regarding the light photosensitivity assay. We thank Asuka Fry and Patrizia Belleda (University Hospital Zurich) for CD34+ cell isolation. We thank Sylvain Egloff for performing preliminary CRISPR experiments and J. Philipp Becker for assistance with cell culture. This research was supported in part by NCCR RNA & Disease, funded by the Swiss National Science Foundation, and by ETH Z?rich. A.P.A.T. is supported by the Professor Dr. Max Clo?tta Foundation. F.H. and J.H. conceived the project; M.D.R. and D.S. designed the CRISPR-Cas9 FECH construct; P.I. characterized the cells and performed compound screening; F.H. conducted photodamage assays, qRT-PCR experiments, and experiments with primary cultures, supported by V.L. and N.V.v.W.; J.B.A. E.M. A.M. X.S.Y. and A.T. designed primary cell experiments; J.J. conducted enzymatic assays; A.G. conducted microscopy and western blotting experiments; J.B.A. and C.P.G. performed PPIX quantification assays. F.H. D.S. and J.H. wrote the manuscript. J.B.A. is a founding member and vice president of the International Porphyria Patient Network, a not-for-profit patient organization. A.M. received funding from Clinuvel Pharmaceuticals for research on EPP. Funding Information: J.B.A. is a founding member and vice president of the International Porphyria Patient Network, a not-for-profit patient organization. A.M. received funding from Clinuvel Pharmaceuticals for research on EPP. Publisher Copyright: {\textcopyright} 2021 Elsevier Ltd Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",

year = "2021",

month = aug,

day = "19",

doi = "10.1016/j.chembiol.2021.02.021",

language = "English",

volume = "28",

pages = "1221--1234.e6",

journal = "Cell Chemical Biology",

issn = "2451-9456",

publisher = "Elsevier Inc.",

number = "8",

}

Halloy, F, Iyer, PS, Ghidini, A, Lysenko, V, Barman-Aksözen, J, Grubenmann, CP, Jucker, J, Wildner-Verhey van Wijk, N, Ruepp, MD, Minder, EI, Minder, AE, Schneider-Yin, X, Theocharides, APA, Schümperli, D & Hall, J 2021, 'Repurposing of glycine transport inhibitors for the treatment of erythropoietic protoporphyria', Cell Chemical Biology, vol. 28, no. 8, pp. 1221-1234.e6. https://doi.org/10.1016/j.chembiol.2021.02.021

TY - JOUR

T1 - Repurposing of glycine transport inhibitors for the treatment of erythropoietic protoporphyria

AU - Halloy, François

AU - Iyer, Pavithra S.

AU - Ghidini, Alice

AU - Lysenko, Veronika

AU - Barman-Aksözen, Jasmin

AU - Grubenmann, Chia Pei

AU - Jucker, Jessica

AU - Wildner-Verhey van Wijk, Nicole

AU - Ruepp, Marc David

AU - Minder, Elisabeth I.

AU - Minder, Anna Elisabeth

AU - Schneider-Yin, Xiaoye

AU - Theocharides, Alexandre P.A.

AU - Schümperli, Daniel

AU - Hall, Jonathan

N1 - Funding Information: We thank Dr. Malgorzata Kisielow (ETH Zurich, Flow Cytometry Core Facility) for useful input and discussions on PPIX quantification by cytofluorometry. We thank André Imboden (ETH Zurich, Department of Biology) for technical lamp information regarding the light photosensitivity assay. We thank Asuka Fry and Patrizia Belleda (University Hospital Zurich) for CD34 + cell isolation. We thank Sylvain Egloff for performing preliminary CRISPR experiments and J. Philipp Becker for assistance with cell culture. This research was supported in part by NCCR RNA & Disease , funded by the Swiss National Science Foundation , and by ETH Zürich . A.P.A.T. is supported by the Professor Dr. Max Cloëtta Foundation . Funding Information: We thank Dr. Malgorzata Kisielow (ETH Zurich, Flow Cytometry Core Facility) for useful input and discussions on PPIX quantification by cytofluorometry. We thank Andr? Imboden (ETH Zurich, Department of Biology) for technical lamp information regarding the light photosensitivity assay. We thank Asuka Fry and Patrizia Belleda (University Hospital Zurich) for CD34+ cell isolation. We thank Sylvain Egloff for performing preliminary CRISPR experiments and J. Philipp Becker for assistance with cell culture. This research was supported in part by NCCR RNA & Disease, funded by the Swiss National Science Foundation, and by ETH Z?rich. A.P.A.T. is supported by the Professor Dr. Max Clo?tta Foundation. F.H. and J.H. conceived the project; M.D.R. and D.S. designed the CRISPR-Cas9 FECH construct; P.I. characterized the cells and performed compound screening; F.H. conducted photodamage assays, qRT-PCR experiments, and experiments with primary cultures, supported by V.L. and N.V.v.W.; J.B.A. E.M. A.M. X.S.Y. and A.T. designed primary cell experiments; J.J. conducted enzymatic assays; A.G. conducted microscopy and western blotting experiments; J.B.A. and C.P.G. performed PPIX quantification assays. F.H. D.S. and J.H. wrote the manuscript. J.B.A. is a founding member and vice president of the International Porphyria Patient Network, a not-for-profit patient organization. A.M. received funding from Clinuvel Pharmaceuticals for research on EPP. Funding Information: J.B.A. is a founding member and vice president of the International Porphyria Patient Network, a not-for-profit patient organization. A.M. received funding from Clinuvel Pharmaceuticals for research on EPP. Publisher Copyright: © 2021 Elsevier Ltd Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/8/19

Y1 - 2021/8/19

N2 - Erythropoietic protoporphyria (EPP) is a rare disease in which patients experience severe light sensitivity. It is caused by a deficiency of ferrochelatase (FECH), the last enzyme in heme biosynthesis (HBS). The lack of FECH causes accumulation of its photoreactive substrate protoporphyrin IX (PPIX) in patients' erythrocytes. Here, we explored an approach for the treatment of EPP by decreasing PPIX synthesis using small-molecule inhibitors directed to factors in the HBS pathway. We generated a FECH-knockout clone from K562 erythroleukemia cells, which accumulates PPIX and undergoes oxidative stress upon light exposure. We used these matched cell lines to screen a set of publicly available inhibitors of factors in the HBS pathway. Inhibitors of the glycine transporters GlyT1 and GlyT2 lowered levels of PPIX and markers of oxidative stress selectively in K56211B4 cells, and in primary erythroid cultures from an EPP patient. Our findings open the door to investigation of glycine transport inhibitors for HBS disorders.

AB - Erythropoietic protoporphyria (EPP) is a rare disease in which patients experience severe light sensitivity. It is caused by a deficiency of ferrochelatase (FECH), the last enzyme in heme biosynthesis (HBS). The lack of FECH causes accumulation of its photoreactive substrate protoporphyrin IX (PPIX) in patients' erythrocytes. Here, we explored an approach for the treatment of EPP by decreasing PPIX synthesis using small-molecule inhibitors directed to factors in the HBS pathway. We generated a FECH-knockout clone from K562 erythroleukemia cells, which accumulates PPIX and undergoes oxidative stress upon light exposure. We used these matched cell lines to screen a set of publicly available inhibitors of factors in the HBS pathway. Inhibitors of the glycine transporters GlyT1 and GlyT2 lowered levels of PPIX and markers of oxidative stress selectively in K56211B4 cells, and in primary erythroid cultures from an EPP patient. Our findings open the door to investigation of glycine transport inhibitors for HBS disorders.

KW - CRISPR-Cas9

KW - drug repurposing

KW - erythropoietic protoporphyria

KW - glycine transporter 2

KW - oxidative stress

KW - photodamage

KW - protoporphyrin IX

UR - http://www.scopus.com/inward/record.url?scp=85112706575&partnerID=8YFLogxK

U2 - 10.1016/j.chembiol.2021.02.021

DO - 10.1016/j.chembiol.2021.02.021

M3 - Article

C2 - 33756123

AN - SCOPUS:85112706575

SN - 2451-9456

VL - 28

SP - 1221-1234.e6

JO - Cell Chemical Biology

JF - Cell Chemical Biology

IS - 8

ER -

Repurposing of glycine transport inhibitors for the treatment of erythropoietic protoporphyria

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Keywords

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