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Annexin Animal Models—From Fundamental Principles to Translational Research

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Annexin Animal Models—From Fundamental Principles to Translational Research International Journal of Molecular Sciences Review Annexin Animal Models—From Fundamental Principles to Translational Research Thomas Grewal 1,* , Carles Rentero 2,3 , Carlos Enrich 2,3 , Mohamed Wahba 1, Carsten A. Raabe 4 and Ursula Rescher 4,* 1 School of Pharmacy, Faculty of Medicine and Health, University of Sydney, Sydney, NSW 2006, Australia; [email protected] 2 Departament de Biomedicina, Unitat de Biologia Cel·lular, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, 08036 Barcelona, Spain; [email protected] (C.R.); [email protected] (C.E.) 3 Centre de Recerca Biomèdica CELLEX, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain 4 Research Group Regulatory Mechanisms of Inflammation, Center for Molecular Biology of Inflammation (ZMBE) and Cells in Motion Interfaculty Center (CiM), Institute of Medical Biochemistry, University of Muenster, 48149 Muenster, Germany; [email protected] * Correspondence: [email protected] (T.G.); [email protected] (U.R.); Tel.: +61-(0)2-9351-8496 (T.G.); +49-(0)251-83-52121 (U.R.) Abstract: Routine manipulation of the mouse genome has become a landmark in biomedical research. Traits that are only associated with advanced developmental stages can now be investigated within a living organism, and the in vivo analysis of corresponding phenotypes and functions advances the translation into the clinical setting. The annexins, a family of closely related calcium (Ca2+)- and lipid- binding proteins, are found at various intra- and extracellular locations, and interact with a broad range of membrane lipids and proteins. Their impacts on cellular functions has been extensively Citation: Grewal, T.; Rentero, C.; Enrich, C.; Wahba, M.; Raabe, C.A.; assessed in vitro, yet annexin-deficient mouse models generally develop normally and do not display Rescher, U. Annexin Animal obvious phenotypes. Only in recent years, studies examining genetically modified annexin mouse Models—From Fundamental models which were exposed to stress conditions mimicking human disease often revealed striking Principles to Translational Research. phenotypes. This review is the first comprehensive overview of annexin-related research using Int. J. Mol. Sci. 2021, 22, 3439. animal models and their exciting future use for relevant issues in biology and experimental medicine. https://doi.org/10.3390/ ijms22073439 Keywords: annexins; calcium; human disease models; KO mice; membrane trafficking; membrane organization; extracellular annexin functions Academic Editor: Georg R. Zoidl Received: 22 February 2021 Accepted: 24 March 2021 1. Introduction Published: 26 March 2021 The assembly of the human genome, followed by DNA deep sequencing, changed Publisher’s Note: MDPI stays neutral our approach to analyse gene functions. Genetic tools, which helped to develop a major with regard to jurisdictional claims in part of our understanding of modern biology, started off with phenotypes and aimed to published maps and institutional affil- identify the underlying gene or its allelic variation to infer function. The completion of iations. the human genome assembly marked the onset of methods that build on reverse genetics. Genes and their genomic sequence became available and the generation of mouse models with gene deletions or targeted mutations allowed the in vivo analysis of corresponding phenotypes and functions. Mouse models lacking individual members of the annexin gene family have been Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. established and unravelled some of their in vivo functions. There are many excellent re- This article is an open access article views on the annexin family covering the common and specific characteristics of individual distributed under the terms and annexin family members [1–13]. Hence, in the following, we provide only a brief overview conditions of the Creative Commons necessary to appreciate the generation and functional implications from the in vivo analysis Attribution (CC BY) license (https:// of the annexin mouse models. creativecommons.org/licenses/by/ The annexin family consists of twelve evolutionary conserved and structurally related 2+ 4.0/). Ca - and phospholipid-binding proteins [1,14]. All annexins are structurally related, Int. J. Mol. Sci. 2021, 22, 3439. https://doi.org/10.3390/ijms22073439 https://www.mdpi.com/journal/ijms Int. J. Mol. Sci. 2021, 22, 3439 Int. J. Mol. Sci. 2021, 22, x FOR PEER REVIEW 2 of 67 2 of 60 they encode a variableThe N-terminal annexin family domain, consists which of twelve is responsible evolutionary for conserved different, and annexin structurally re- lated Ca2+- and phospholipid-binding proteins [1,14]. All annexins are structurally related, member-specific functions,they encode and a variable a highly N-terminal conserved domain, C-terminal which is core. responsible The C-terminal for different, do- annexin 2+ main is composed ofmember-specific four highly related functions, repeats and a (eighthighly inconserved case of C-terminal AnxA6) containing core. The C-terminal Ca do- binding sites that enablemain is the composed interaction of four with highly negatively related repeats charged (eight phospholipids in case of AnxA6) (Figure containing1). Ca2+ Extensive in vitro analysisbinding sites solved that theenable three-dimensional the interaction with structures negatively ofcharged annexins phospholipids ([15–18], (Figure reviewed in [2,19,201).]), Extensive their Ca 2+in-dependent vitro analysis affinities solved the to three-dimensional various phospholipids structures and of annexins other ([15– interaction partners18], [1, 2reviewed]. A large in [2,19,20]), number oftheir regulatory Ca2+-dependent roles affinities associated to various withindividual phospholipids and annexins can be explainedother interaction by their partners Ca2+-inducible [1,2]. A large and number rapid translocation of regulatory roles from associated the cytosol with indi- vidual annexins can be explained by their Ca2+-inducible and rapid translocation from the to the plasma membranecytosol or to intracellularthe plasma membrane membranes or intracellular [1–4]. At membranes these locations, [1–4]. At their these unique locations, their N-termini interact withunique dissimilar N-termini binding interact with partners, dissimilar which binding in part partners, explains which the in functional part explains the diversity of annexins.functional In addition, diversity theof annexins. composition In addition and, distributionthe composition of and phospholipids distribution of phos- and other lipids, suchpholipids as phosphatidylinositol-4,5-bisphosphate, and other lipids, such as phosphatidylinositol-4,5-bisphosphate, cholesterol and ceramide cholesterol and within specific membraneceramide microdomains,within specific membrane but also microdomains, posttranslational but also modifications posttranslational and modifica- 2+ pH, contribute to differenttions and spatiotemporal pH, contribute to and different Ca2+ spatiotemporal-sensitive membrane and Ca -sensitive binding membrane kinetics bind- ing kinetics leading to further functional diversification of individual family members [1– leading to further functional diversification of individual family members [1–9,11]. 9,11]. FigureFigure 1. Annexin 1. Annexin structural structural organization. organization. (A) Schematic (A) Schematic overview overviewof annexins. of Mage annexins.nta, N-terminal Magenta, tails; N-terminal light to dark green,tails; C-terminal light to darkcore domains green, with C-terminal Annexin corerepeats domains 1–4 and 5–8 with for AnnexinAnxA6; grey, repeats short spac 1–4er and regions 5–8 between for AnxA6; tail and first Annexin repeat or between Annexin repeats; blue, AnxA6 linker region. (B) 3D-structures of human AnxA1 (PDB: 1AINgrey, [17]) short and spacer(C) bovine regions AnxA6 between (PDB: 1AVC tail [18]) and cores first (light Annexin to dark repeat green, spacer or between regions Annexin in grey, AnxA6 repeats; linker blue, region inAnxA6 blue), with linker coordinated region. ( Bcalcium) 3D-structures ions (red). of 3D-structures human AnxA1 are (PDB:visualized 1AIN with [17 the]) and iCn3D (C) software bovine AnxA6vs. 2.24.6; (https://www.ncbi.nlm.nih.gov(PDB: 1AVC [18]) cores (light/Structure/icn3d/icn3d.htm to dark green, spacerl; accessed regions 12 March in grey, 2021). AnxA6 linker region in blue), with coordinated calcium ions (red). 3D-structures are visualized with the iCn3D software vs. 2.24.6; These processes are thought to control basic cellular functions that are pivotal to the (https://www.ncbi.nlm.nih.gov/Structure/icn3d/icn3d.htmlregulation of growth, development, progra;mmed accessed cell 12 death, March but 2021). also cell motility, mem- brane repair and inflammatory response [1–13]. Ectopic overexpression or siRNA-medi- These processesated are knockdown thought delivered to control great basic insights cellular in the functionsannexin biochemistry that are pivotaland broadened to our the regulation of growth,knowledge development, substantially. More programmed advanced technologies cell death, of but reverse also genetics cell motility, made knockout membrane repair and inflammatory response [1–13]. Ectopic overexpression or siRNA- mediated knockdown delivered great insights in the annexin biochemistry and broadened our knowledge substantially. More advanced technologies of reverse genetics made knock- out (KO) mouse models the gold standard to analyse annexin functions.
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