Age-related macular degeneration associated polymorphism rs10490924 in ARMS2 results in deficiency of a complement activator

Sven Micklisch; Yuchen Lin; Saskia Jacob; Marcus Karlstetter; Katharina Dannhausen; Prasad Dasari; Monika von der Heide; Hans Martin Dahse; Lisa Schmölz; Felix Grassmann; Medhanie Alene; Sascha Fauser; Harald Neumann; Stefan Lorkowski; Diana Pauly; Bernhard H. Weber; Antonia M. Joussen; Thomas Langmann; Peter F. Zipfel; Christine Skerka

doi:10.1186/s12974-016-0776-3

Age-related macular degeneration associated polymorphism rs10490924 in ARMS2 results in deficiency of a complement activator

Sven Micklisch, Yuchen Lin, Saskia Jacob, Marcus Karlstetter, Katharina Dannhausen, Prasad Dasari, Monika von der Heide, Hans Martin Dahse, Lisa Schmölz, Felix Grassmann, Medhanie Alene, Sascha Fauser, Harald Neumann, Stefan Lorkowski, Diana Pauly, Bernhard H. Weber, Antonia M. Joussen, Thomas Langmann, Peter F. Zipfel, Christine Skerka^*

^*Corresponding author for this work

Medical Sciences

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Abstract

Background: Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. The polymorphism rs10490924 in the ARMS2 gene is highly associated with AMD and linked to an indel mutation (del443ins54), the latter inducing mRNA instability. At present, the function of the ARMS2 protein, the exact cellular sources in the retina and the biological consequences of the rs10490924 polymorphism are unclear. Methods: Recombinant ARMS2 was expressed in Pichia pastoris, and protein functions were studied regarding cell surface binding and complement activation in human serum using fluoresence-activated cell sorting (FACS) as well as laser scanning microscopy (LSM). Biolayer interferometry defined protein interactions. Furthermore, endogenous ARMS2 gene expression was studied in human blood derived monocytes and in human induced pluripotent stem cell-derived microglia (iPSdM) by PCR and LSM. The ARMS2 protein was localized in human genotyped retinal sections and in purified monocytes derived from AMD patients without the ARMS2 risk variant by LSM. ARMS2 expression in monocytes under oxidative stress was determined by Western blot analysis. Results: Here, we demonstrate for the first time that ARMS2 functions as surface complement regulator. Recombinant ARMS2 binds to human apoptotic and necrotic cells and initiates complement activation by recruiting the complement activator properdin. ARMS2-properdin complexes augment C3b surface opsonization for phagocytosis. We also demonstrate for the first time expression of ARMS2 in human monocytes especially under oxidative stress and in microglia cells of the human retina. The ARMS2 protein is absent in monocytes and also in microglia cells, derived from patients homozygous for the ARMS2 AMD risk variant (rs10490924). Conclusions: ARMS2 is likely involved in complement-mediated clearance of cellular debris. As AMD patients present with accumulated proteins and lipids on Bruch's membrane, ARMS2 protein deficiency due to the genetic risk variant might be involved in drusen formation.

Original language	English
Article number	4
Number of pages	15
Journal	Journal of Neuroinflammation
Volume	14
Early online date	5 Jan 2017
DOIs	https://doi.org/10.1186/s12974-016-0776-3
Publication status	Published - 5 Jan 2017

Bibliographical note

Acknowledgements: The authors thank all German AMD patients for their participation. We also thank Maria Pötsch (Leibniz Institute for Natural Product Reseach and Infection Biology, Jena) for MS analyses.

Funding: This research was supported by the German Council “Deutsche Forschungs-Gemeinschaft” SK46, Zi432, LA1206, the “Pro Retina” foundation and the Thuringian Ministry of Science and Education, Germany. HN is a member of the DFG-funded excellence cluster ImmunoSensation (EXC 1023). YL is a doctoral researcher at the International Leibniz Research School (ILRS), part of the Jena school of Microbial Communication (JSMC).

Availability of data and materials: Materials are available at christine.skerka@hki-jena.de.

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Micklisch, S., Lin, Y., Jacob, S., Karlstetter, M., Dannhausen, K., Dasari, P., von der Heide, M., Dahse, H. M., Schmölz, L., Grassmann, F., Alene, M., Fauser, S., Neumann, H., Lorkowski, S., Pauly, D., Weber, B. H., Joussen, A. M., Langmann, T., Zipfel, P. F., & Skerka, C. (2017). Age-related macular degeneration associated polymorphism rs10490924 in ARMS2 results in deficiency of a complement activator. Journal of Neuroinflammation, 14, Article 4. https://doi.org/10.1186/s12974-016-0776-3

Micklisch, S, Lin, Y, Jacob, S, Karlstetter, M, Dannhausen, K, Dasari, P, von der Heide, M, Dahse, HM, Schmölz, L, Grassmann, F, Alene, M, Fauser, S, Neumann, H, Lorkowski, S, Pauly, D, Weber, BH, Joussen, AM, Langmann, T, Zipfel, PF & Skerka, C 2017, 'Age-related macular degeneration associated polymorphism rs10490924 in ARMS2 results in deficiency of a complement activator', Journal of Neuroinflammation, vol. 14, 4. https://doi.org/10.1186/s12974-016-0776-3

@article{00f05afb703d44f2b4f6058792c355ca,

title = "Age-related macular degeneration associated polymorphism rs10490924 in ARMS2 results in deficiency of a complement activator",

abstract = "Background: Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. The polymorphism rs10490924 in the ARMS2 gene is highly associated with AMD and linked to an indel mutation (del443ins54), the latter inducing mRNA instability. At present, the function of the ARMS2 protein, the exact cellular sources in the retina and the biological consequences of the rs10490924 polymorphism are unclear. Methods: Recombinant ARMS2 was expressed in Pichia pastoris, and protein functions were studied regarding cell surface binding and complement activation in human serum using fluoresence-activated cell sorting (FACS) as well as laser scanning microscopy (LSM). Biolayer interferometry defined protein interactions. Furthermore, endogenous ARMS2 gene expression was studied in human blood derived monocytes and in human induced pluripotent stem cell-derived microglia (iPSdM) by PCR and LSM. The ARMS2 protein was localized in human genotyped retinal sections and in purified monocytes derived from AMD patients without the ARMS2 risk variant by LSM. ARMS2 expression in monocytes under oxidative stress was determined by Western blot analysis. Results: Here, we demonstrate for the first time that ARMS2 functions as surface complement regulator. Recombinant ARMS2 binds to human apoptotic and necrotic cells and initiates complement activation by recruiting the complement activator properdin. ARMS2-properdin complexes augment C3b surface opsonization for phagocytosis. We also demonstrate for the first time expression of ARMS2 in human monocytes especially under oxidative stress and in microglia cells of the human retina. The ARMS2 protein is absent in monocytes and also in microglia cells, derived from patients homozygous for the ARMS2 AMD risk variant (rs10490924). Conclusions: ARMS2 is likely involved in complement-mediated clearance of cellular debris. As AMD patients present with accumulated proteins and lipids on Bruch's membrane, ARMS2 protein deficiency due to the genetic risk variant might be involved in drusen formation.",

author = "Sven Micklisch and Yuchen Lin and Saskia Jacob and Marcus Karlstetter and Katharina Dannhausen and Prasad Dasari and {von der Heide}, Monika and Dahse, {Hans Martin} and Lisa Schm{\"o}lz and Felix Grassmann and Medhanie Alene and Sascha Fauser and Harald Neumann and Stefan Lorkowski and Diana Pauly and Weber, {Bernhard H.} and Joussen, {Antonia M.} and Thomas Langmann and Zipfel, {Peter F.} and Christine Skerka",

note = "Acknowledgements: The authors thank all German AMD patients for their participation. We also thank Maria P{\"o}tsch (Leibniz Institute for Natural Product Reseach and Infection Biology, Jena) for MS analyses. Funding: This research was supported by the German Council “Deutsche Forschungs-Gemeinschaft” SK46, Zi432, LA1206, the “Pro Retina” foundation and the Thuringian Ministry of Science and Education, Germany. HN is a member of the DFG-funded excellence cluster ImmunoSensation (EXC 1023). YL is a doctoral researcher at the International Leibniz Research School (ILRS), part of the Jena school of Microbial Communication (JSMC). Availability of data and materials: Materials are available at christine.skerka@hki-jena.de.",

year = "2017",

month = jan,

day = "5",

doi = "10.1186/s12974-016-0776-3",

language = "English",

volume = "14",

journal = "Journal of Neuroinflammation",

issn = "1742-2094",

publisher = "BioMed Central",

}

TY - JOUR

T1 - Age-related macular degeneration associated polymorphism rs10490924 in ARMS2 results in deficiency of a complement activator

AU - Micklisch, Sven

AU - Lin, Yuchen

AU - Jacob, Saskia

AU - Karlstetter, Marcus

AU - Dannhausen, Katharina

AU - Dasari, Prasad

AU - von der Heide, Monika

AU - Dahse, Hans Martin

AU - Schmölz, Lisa

AU - Grassmann, Felix

AU - Alene, Medhanie

AU - Fauser, Sascha

AU - Neumann, Harald

AU - Lorkowski, Stefan

AU - Pauly, Diana

AU - Weber, Bernhard H.

AU - Joussen, Antonia M.

AU - Langmann, Thomas

AU - Zipfel, Peter F.

AU - Skerka, Christine

N1 - Acknowledgements: The authors thank all German AMD patients for their participation. We also thank Maria Pötsch (Leibniz Institute for Natural Product Reseach and Infection Biology, Jena) for MS analyses. Funding: This research was supported by the German Council “Deutsche Forschungs-Gemeinschaft” SK46, Zi432, LA1206, the “Pro Retina” foundation and the Thuringian Ministry of Science and Education, Germany. HN is a member of the DFG-funded excellence cluster ImmunoSensation (EXC 1023). YL is a doctoral researcher at the International Leibniz Research School (ILRS), part of the Jena school of Microbial Communication (JSMC). Availability of data and materials: Materials are available at christine.skerka@hki-jena.de.

PY - 2017/1/5

Y1 - 2017/1/5

N2 - Background: Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. The polymorphism rs10490924 in the ARMS2 gene is highly associated with AMD and linked to an indel mutation (del443ins54), the latter inducing mRNA instability. At present, the function of the ARMS2 protein, the exact cellular sources in the retina and the biological consequences of the rs10490924 polymorphism are unclear. Methods: Recombinant ARMS2 was expressed in Pichia pastoris, and protein functions were studied regarding cell surface binding and complement activation in human serum using fluoresence-activated cell sorting (FACS) as well as laser scanning microscopy (LSM). Biolayer interferometry defined protein interactions. Furthermore, endogenous ARMS2 gene expression was studied in human blood derived monocytes and in human induced pluripotent stem cell-derived microglia (iPSdM) by PCR and LSM. The ARMS2 protein was localized in human genotyped retinal sections and in purified monocytes derived from AMD patients without the ARMS2 risk variant by LSM. ARMS2 expression in monocytes under oxidative stress was determined by Western blot analysis. Results: Here, we demonstrate for the first time that ARMS2 functions as surface complement regulator. Recombinant ARMS2 binds to human apoptotic and necrotic cells and initiates complement activation by recruiting the complement activator properdin. ARMS2-properdin complexes augment C3b surface opsonization for phagocytosis. We also demonstrate for the first time expression of ARMS2 in human monocytes especially under oxidative stress and in microglia cells of the human retina. The ARMS2 protein is absent in monocytes and also in microglia cells, derived from patients homozygous for the ARMS2 AMD risk variant (rs10490924). Conclusions: ARMS2 is likely involved in complement-mediated clearance of cellular debris. As AMD patients present with accumulated proteins and lipids on Bruch's membrane, ARMS2 protein deficiency due to the genetic risk variant might be involved in drusen formation.

AB - Background: Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. The polymorphism rs10490924 in the ARMS2 gene is highly associated with AMD and linked to an indel mutation (del443ins54), the latter inducing mRNA instability. At present, the function of the ARMS2 protein, the exact cellular sources in the retina and the biological consequences of the rs10490924 polymorphism are unclear. Methods: Recombinant ARMS2 was expressed in Pichia pastoris, and protein functions were studied regarding cell surface binding and complement activation in human serum using fluoresence-activated cell sorting (FACS) as well as laser scanning microscopy (LSM). Biolayer interferometry defined protein interactions. Furthermore, endogenous ARMS2 gene expression was studied in human blood derived monocytes and in human induced pluripotent stem cell-derived microglia (iPSdM) by PCR and LSM. The ARMS2 protein was localized in human genotyped retinal sections and in purified monocytes derived from AMD patients without the ARMS2 risk variant by LSM. ARMS2 expression in monocytes under oxidative stress was determined by Western blot analysis. Results: Here, we demonstrate for the first time that ARMS2 functions as surface complement regulator. Recombinant ARMS2 binds to human apoptotic and necrotic cells and initiates complement activation by recruiting the complement activator properdin. ARMS2-properdin complexes augment C3b surface opsonization for phagocytosis. We also demonstrate for the first time expression of ARMS2 in human monocytes especially under oxidative stress and in microglia cells of the human retina. The ARMS2 protein is absent in monocytes and also in microglia cells, derived from patients homozygous for the ARMS2 AMD risk variant (rs10490924). Conclusions: ARMS2 is likely involved in complement-mediated clearance of cellular debris. As AMD patients present with accumulated proteins and lipids on Bruch's membrane, ARMS2 protein deficiency due to the genetic risk variant might be involved in drusen formation.

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JO - Journal of Neuroinflammation

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Age-related macular degeneration associated polymorphism rs10490924 in ARMS2 results in deficiency of a complement activator

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