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Functional analysis of androglobin, a novel oxygen-binding protein associated with male infertility.

English title Functional analysis of androglobin, a novel oxygen-binding protein associated with male infertility.
Applicant Hoogewijs David
Number 173000
Funding scheme Project funding
Research institution Division de Physiologie Département de Médecine Université de Fribourg
Institution of higher education University of Fribourg - FR
Main discipline Physiology : other topics
Start/End 01.09.2017 - 28.02.2022
Approved amount 402'000.00
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All Disciplines (4)

Discipline
Physiology : other topics
Biophysics
Molecular Biology
Biochemistry

Keywords (4)

oxygen; spermatogenesis; hypoxia; globin

Lay Summary (German)

Lead
David Hoogewijs
Lay summary

Hämoglobin und Myoglobin sind die beiden am besten erforschten sauerstoffbindenden Atmungsproteine. Die postgenomische Ära hat ergeben, dass zusätzliche Globintypen in Säugetieren vorhanden sind, darunter das Neuroglobin und Cytoglobin. Vor kurzem konnten wir eine neuartige, stark evolutionär konservierte Globinklasse identifizieren. Diese umfasst grosse,chimäre Proteinen mit einer N-terminalen Proteasedomäne und einer zentralen Globindomäne. Aufgrund ihrer starken Expression im Hodengewebe werden die Proteine als Androglobin (Adgb) bezeichnet. Die präliminäre Charakterisierung eines neu generierten Adgb-defizienten Mausmodells zeigt als Phänotyp eine Entwicklungsstörung während der Spermatogenese. Das Projekt wird die physiologische Rolle, die biomedizinischen Implikationen und die Genregulation von Adgb in vivo und in vitro umfassend erforschen. Auf der Grundlage des Unfruchtbarkeitsphänotyps im Maus-Knockout-Modell konzentrieren wir uns zunächst auf die Hoden-spezifische Rolle von Adgb. Dabei werden Hoden und Spermien histologisch untersucht, die Sperma-Funktionalität getestet und auf molekularer Ebene Analysen des Transkriptoms und des Interaktoms durchgeführt. Biophysikalische und biochemische Studien sollen hypothetische enzymatische Funktionen oder eine signalvermittelnde Rolle von Adgb aufklären. In Zellkulturen mit gentechnisch modulierter Adgb-Expression wird eine potenzielle anti-oxidative Rolle von Adgb untersucht. Darüber hinaus werden wir die subzelluläre Lokalisation von Adgb bestimmen und die physikalisch-chemischen Eigenschaften des rekombinanten Proteins charakterisieren. Die Genregulation von Adgb wird durch Reportergen-Assays und epigenetische Ansätze analysiert, um Veränderungen des Expressionsniveaus unter pathologischen Bedingungen zu verstehen. Zusammenfassend werden unsere Studien sehr wahrscheinlich zu einem besseren molekularen Verständnis der männlichen Unfruchtbarkeit beitragen.

Direct link to Lay Summary Last update: 04.08.2017

Responsible applicant and co-applicants

Employees

Project partner

Publications

Publication
A reliable set of reference genes to normalize oxygen-dependent cytoglobin gene expression levels in melanoma
De Backer Joey, Maric Darko, Bosman Matthias, Dewilde Sylvia, Hoogewijs David (2021), A reliable set of reference genes to normalize oxygen-dependent cytoglobin gene expression levels in melanoma, in Scientific Reports, 11(1), 10879-10879.
Androglobin gene expression patterns and FOXJ1-dependent regulation indicate its functional association with ciliogenesis
Koay Teng Wei, Osterhof Carina, Orlando Ilaria M.C., Keppner Anna, Andre Daniel, Yousefian Schayan, Alonso María Suárez, Correia Miguel, Markworth Robert, Schödel Johannes, Hankeln Thomas, Hoogewijs David (2021), Androglobin gene expression patterns and FOXJ1-dependent regulation indicate its functional association with ciliogenesis, in Journal of Biological Chemistry, 100291-100291.
Structural modeling of a novel membrane-bound globin-coupled sensor in Geobacter sulfurreducens
Hammerschmid Dietmar, Germani Francesca, Drusin Salvador I., Fagnen Charline, Schuster Claudio D., Hoogewijs David, Marti Marcelo A., Venien-Bryan Catherine, Moens Luc, Van Doorslaer Sabine, Sobott Frank, Dewilde Sylvia (2021), Structural modeling of a novel membrane-bound globin-coupled sensor in Geobacter sulfurreducens, in Computational and Structural Biotechnology Journal, 19, 1874-1888.
Lessons from the post-genomic era: Globin diversity beyond oxygen binding and transport
Keppner Anna, Maric Darko, Correia Miguel, Koay Teng Wei, Orlando Ilaria M.C., Vinogradov Serge N., Hoogewijs David (2020), Lessons from the post-genomic era: Globin diversity beyond oxygen binding and transport, in Redox Biology, 37, 101687-101687.
The Antioxidative Role of Cytoglobin in Podocytes: Implications for a Role in Chronic Kidney Disease
Randi Elisa B., Vervaet Benjamin, Tsachaki Maria, Porto Elena, Vermeylen Stijn, Lindenmeyer Maja T., Thuy Le Thi Thanh, Cohen Clemens D., Devuyst Olivier, Kistler Andreas D., Szabo Csaba, Kawada Norifumi, Hankeln Thomas, Odermatt Alex, Dewilde Sylvia, Wenger Roland H., Hoogewijs David (2020), The Antioxidative Role of Cytoglobin in Podocytes: Implications for a Role in Chronic Kidney Disease, in Antioxidants & Redox Signaling, 32(16), 1155-1171.
Deletion of the serine protease CAP2/Tmprss4 leads to dysregulated renal water handling upon dietary potassium depletion
Keppner Anna, Maric Darko, Sergi Chloé, Ansermet Camille, De Bellis Damien, Kratschmar Denise V., Canonica Jérémie, Klusonova Petra, Fenton Robert A., Odermatt Alex, Crambert Gilles, Hoogewijs David, Hummler Edith (2019), Deletion of the serine protease CAP2/Tmprss4 leads to dysregulated renal water handling upon dietary potassium depletion, in Scientific Reports, 9(1), 19540-19540.
A search for molecular mechanisms underlying male idiopathic infertility
Bracke An, Peeters Kris, Punjabi Usha, Hoogewijs David, Dewilde Sylvia (2018), A search for molecular mechanisms underlying male idiopathic infertility, in Reproductive BioMedicine Online, 36(3), 327-339.
Exploring three different expression systems for recombinant expression of globins: Escherichia coli, Pichia pastoris and Spodoptera frugiperda
Bracke An, Hoogewijs David, Dewilde Sylvia (2018), Exploring three different expression systems for recombinant expression of globins: Escherichia coli, Pichia pastoris and Spodoptera frugiperda, in Analytical Biochemistry, 543, 62-70.
CRISPR activator approaches to study endogenous androglobin gene regulation
KoayTeng Wei, SchödelJohannes, HoogewijsDavid, CRISPR activator approaches to study endogenous androglobin gene regulation, in Methods in Molecular Biology.

Collaboration

Group / person Country
Types of collaboration
University of Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
University of Erlangen Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
University of Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure
University of Dresden Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
University of Basel Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
University of Rennes France (Europe)
- in-depth/constructive exchanges on approaches, methods or results

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Fribourg 10th Research day in Medicine Poster On the function of androglobin: linking oxygen physiology and ciliogenesis. 23.03.2022 Fribourg, Switzerland Hoogewijs David; Keppner Anna;
Fribourg 10th Research day in Medicine Poster Exploring the protective effects of androglobin in cellular models 23.03.2022 Fribourg, Switzerland Hoogewijs David; Keppner Anna;
Fribourg 10th Research day in Medicine Poster Androglobin knockout leads to dysfunctional aldosterone production 23.03.2022 Fribourg, Switzerland Hoogewijs David; Keppner Anna;
100th annual meeting of the German Physiological Society Talk given at a conference On the function of androglobin: linking oxygen physiology and ciliogenesis 02.10.2021 Frankfurt, Germany Koay Teng Wei; Keppner Anna; Hoogewijs David;
100th annual meeting of the German Physiological Society Poster Calcium-mediated proteolysis of androglobin 01.10.2021 Frankfurt, Germany Koay Teng Wei; Hoogewijs David;
100th annual meeting of the German Physiological Society Talk given at a conference Study of the function of androglobin in renal ciliogenesis 30.09.2021 Frankfurt, Germany Keppner Anna; Hoogewijs David;
Swiss Physiological Society Annual meeting Poster Genetic ablation of the newly identified androglobin leads to multiple phenotypes 06.09.2021 Fribourg, Switzerland Keppner Anna; Koay Teng Wei; Hoogewijs David;
Swiss Physiological Society Annual meeting Talk given at a conference Calcium-mediated proteolysis of androglobin 06.09.2021 Fribourg, Switzerland Hoogewijs David; Koay Teng Wei;
Fribourg 9th Research day in Medicine Poster Calcium-mediated processing of androglobin. 14.04.2021 Fribourg (online), Switzerland Koay Teng Wei; Hoogewijs David;
Fribourg 9th Research day in Medicine Talk given at a conference Genetic ablation of the newly identified androglobin leads to multiple phenotypes 14.04.2021 Fribourg (online), Switzerland Keppner Anna; Koay Teng Wei; Hoogewijs David;
LS2 annual meeting Poster Unraveling the cryptic transcriptional landscape of the androglobin gene 17.02.2021 Geneva (online), Switzerland Koay Teng Wei; Hoogewijs David; Keppner Anna;
LS2 Annual meeting Poster Absence of androglobin leads to male infertility 17.02.2021 Geneva (online), Switzerland Keppner Anna; Koay Teng Wei; Hoogewijs David;
LS2 annual meeting Poster Expression of androglobin in the female reproductive tract 13.02.2020 Zürich, Switzerland Hoogewijs David;
Annual meeting of the German Physiological Society (DPG) Poster Genetic ablation of the newly identified androglobin leads to multiple phenotypes 01.10.2019 Ulm, Germany Keppner Anna; Hoogewijs David;
Annual meeting of the German Physiological Society (DPG) Poster Transcriptional regulation of androglobin and calcium-mediated androglobin cleavage 01.10.2019 Ulm, Germany Hoogewijs David; Koay Teng Wei;
Swiss Physiological Society Annual Meeting Individual talk Genetic ablation of the newly identified androglobin leads to multiple phenotypes 03.09.2019 Bern, Switzerland, Switzerland Keppner Anna; Hoogewijs David;
Swiss Physiological Society Annual Meeting Poster Transcriptional regulation of androglobin and calcium-mediated androglobin cleavage. 03.09.2019 Bern, Switzerland Koay Teng Wei; Hoogewijs David;
Europhysiology Poster Androglobin is a recently discovered oxygen-binding globin with a crucial role in spermatogenesis 14.09.2018 London, Great Britain and Northern Ireland Koay Teng Wei; Keppner Anna; Hoogewijs David;
Europhysiology Poster Genetic ablation of androglobin leads to multiple phenotypes 14.09.2018 London, Great Britain and Northern Ireland Hoogewijs David; Keppner Anna;
Europhysiology Poster Transcriptional regulation and activation of a novel globin gene – androglobin 14.09.2018 London, Great Britain and Northern Ireland Koay Teng Wei; Hoogewijs David;
XXth International Conference on Oxygen Binding and Sensing Proteins Poster Genetic ablation of androglobin leads to multiple phenotypes 04.09.2018 Barcelona, Spain Keppner Anna; Hoogewijs David;
XXth International Conference on Oxygen Binding and Sensing Proteins Poster The anti-oxidative role of cytoglobin in podocytes suggests an association with chronic kidney disease 04.09.2018 Barcelona, Spain Hoogewijs David;
Swiss Physiological Society Annual Meeting Poster Androglobin - a novel globin protein: insights into its function and expression 04.09.2018 Fribourg, Switzerland Koay Teng Wei; Hoogewijs David; Keppner Anna;
XXth International Conference on Oxygen Binding and Sensing Proteins Talk given at a conference Genetic ablation of androglobin reveals multiple phenotypes 04.09.2018 Barcelona, Spain Hoogewijs David;
XXth International Conference on Oxygen Binding and Sensing Proteins Poster Androglobin is a recently discovered oxygen-binding globin with a crucial role in spermatogenesis 04.09.2018 Barcelona, Spain Keppner Anna; Hoogewijs David;
Swiss Physiological Society Annual meeting Poster Deciphering the molecular function of androglobin 04.09.2018 Fribourg, Switzerland Hoogewijs David; Koay Teng Wei;
XXth International Conference on Oxygen Binding and Sensing Proteins Poster Expression of androglobin in the female reproductive tract 04.09.2018 Barcelona, Spain Hoogewijs David;
Swiss Society of Pharmacology and Toxicology Annual Meeting Poster Androglobin: a newly identified globin required for male fertility in mice 19.04.2018 Bern, Switzerland Keppner Anna; Hoogewijs David;
LS2 annual meeting Poster Genetic ablation of androglobin leads to multiple phenotypes 12.02.2018 Lausanne, Switzerland Hoogewijs David; Keppner Anna;
COST Action BM1407 meeting Talk given at a conference Genetic ablation of androglobin, the 5th mammalian globin, leads to multiple phenotypes 07.02.2018 Lissabon, Portugal Hoogewijs David;
49th Annual Meeting of the Swiss Society of Nephrology Poster Antioxidative role of cytoglobin in podocytes and its association with chronic kidney disease 07.12.2017 Fribourg, Switzerland Hoogewijs David;
Swiss Physiological Society Annual Meeting Poster Androglobin: a newly identified globin required for male fertility in mice 05.09.2017 Bern, Switzerland Hoogewijs David; Keppner Anna;


Awards

Title Year
Best poster price, Fribourg 10th Research day in Medicine, Fribourg (Switzerland) 2022
Best talk price, Fribourg 9th Research day in Medicine, online Fribourg (Switzerland) 2021
Best talk price, Swiss Physiology society, annual meeting, online Fribourg (Switzerland) 2020
Best poster price, German Physiological Society (DPG) annual meeting (Ulm) 2019
Best talk price, Swiss Physiological Society, annual meeting, Bern (Switzerland) 2019
Best Poster price, Fribourg 7th Research day in Medicine, Fribourg (Switzerland) 2018
Best poster price, Life Sciences Switzerland (LS2) meeting, Lausanne (Switzerland) 2018
Best poster price, XXth International Conference on Oxygen Binding and Sensing Proteins, Barcelona (Spain) 2018
Best poster price, XXth International Conference on Oxygen Binding and Sensing Proteins, Barcelona (Spain) 2018
Best Poster price, Fribourg 6th Research day in Medicine, Fribourg (Switzerland) 2017
Best talk price, Swiss society of Pharmacology and Toxicology, annual meeting, Bern (Switzerland), 2017

Associated projects

Number Title Start Funding scheme
189785 Multichannel confocal microscope with fluorescence lifetime imaging for life science samples 01.03.2020 R'EQUIP
205625 Mouse metabolic phenotyping platform for UNIFR 01.12.2021 R'EQUIP

Abstract

Background: Hemoglobin and myoglobin represent the two most-established and well-studied heme-containing oxygen binding proteins. The post-genomic era revealed the presence of additional mammalian globin types, including neuroglobin and cytoglobin. Recently the applicant and collaborating labs identified a new metazoan globin lineage, consisting of large chimeric proteins with an N-terminal cysteine protease domain and a central globin domain, named androglobins (Adgbs), because of their specific expression in testis tissue (Hoogewijs et al. 2012a). Intriguingly, this new member of the globin family is evolutionary ancient and extremely conserved. We showed that Adgb expression is associated with postmeiotic stages of spermatogenesis. Male infertility of an Adgb-deficient mouse model and higher Adgb expression levels in spermatozoa from fertile vs. infertile males underscore the functional importance of Adgb during spermatogenesis. The present project will analyse the physiological role of Adgb using a variety of in vitro and in vivo approaches with focus on spermatogenesis.Aims: The main goal of the present proposal is to explore the functional role of Adgb. To achieve this goal, in vitro experiments were designed to determine the biochemical properties of Adgb and to characterize the mechanism of Adgb expression regulation. In vivo experiments include the completion of the ongoing generation of a novel conditional Adgb knock-out mouse model and aim to elucidate the role of Adgb during spermatogenesis. Research plan: In order to gain more insights into the function of Adgb we will perform in vitro as well as in vivo experiments. We will use cell culture studies to modify expression levels of Adgb and look at potential downstream targets by applying transcriptome profiling. Functional studies will evaluate if Adgb has a potential protection function against reactive oxygen species (ROS) in these cell models. Additionally, we will explore the transcriptional regulation of Adgb using reporter gene assays and DNA methylation status-modifying agents. Physico-chemical characterization of the recombinantly expressed Adgb globin domain (wild-type and mutants) will be performed to determine the kinetic properties of ligand binding. Functionality of the protease domain will be assessed on the recombinantly expressed calpain domain (wild-type and mutants) and full-length protein. To complement the in vitro studies we will characterize a novel conditional Adgb knock-out mouse model and initially focus on the testis-specific role of Adgb, employing testes and sperm histology, sperm functionality assays, transcriptomics and interactome studies.Significance: The elucidation of the testis-specific function of Adgb is mainly a basic science research project. However, the finding of substantially decreased Adgb expression levels in spermatozoa from infertile males and the infertility of androglobin-deficient mice clearly indicates that it could have important biomedical implications with regard to the increased occurrence of male infertility in our society. Male infertility is affected by oxygenation and ROS, among many other factors, and Adgb might be involved in sensing oxygen levels or detoxification of harmful ROS. Thereby our studies will contribute to an improved understanding of male infertility.
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