Microbial Biochemistry and Physiology (until 2015)

We investigate the pathogenesis and adaptation of fungal pathogens.

  • Nutrient acquisition and metabolism of pathogenic fungi during infection
  • In vivo real-time imaging of fungal infections in murine model systems
  • Secondary metabolites of Aspergillus terreus

In recent years, life-threatening fungal infections have become more important. This is mainly due to increasing numbers in patients under immunosuppressive regimens. Unfortunately, the scientific knowledge on fungal infections, as well as therapeutic strategies are limited.

For this reason, we are investigating the potential of pathogenic fungi to acquire and metabolise host-derived nutrients. In this research, we are looking for metabolic processes common in different fungal species, but also on pathways specific for selected pathogens to resemble the great variety of possible host-pathogen interactions. Since nutrient acquisition is essential for successful host colonisation, the aim of these studies is the definition of new antimycotic targets.

Studies of the infection process frequently use conventional infection models that can only provide snapshots of the infection from which then the broader picture of the disease progression has to be assembled. Therefore, we are developing in vivo imaging systems that allow the visualisation of disease progression in individual living animals in temporal and spatial resolution. This enables the monitoring of antimycotic therapy efficacy under in vivo conditions and permits the detection of cryptic niches of infection that may be overlooked by histological analyses.

Last, but not least, using Aspergillus terreus as a model organism, we investigate the impact of secondary metabolites on fungal pathogenesis and environmental adaptation. Selected metabolites are tested for their biologic activities to draw conclusions on the connection between natural product synthesis and environmental factors.

Head

Dr. habil. Matthias Brock

Now: The University of Nottingham

Topics

Carbon and nitrogen metabolism

Recognition, uptake and utilisation of nutrient sources are an essential prerequisite for all living organisms. Thus, pathogenic microorganisms must not only be able to escape the host immune response, but also to acquire nutrients from the host environment.

In our studies, we investigate the metabolic physiology of pathogenic fungi to elucidate the specific impact of nutritional pathways for the infection process. Thereby, we focus on both catabolic processes and anabolism. Examples are the utilisation of propionyl-CoA generating nutrient sources, the impact of the glyoxylate cycle in virulence and the de novo synthesis of the amino acid lysine. Since essential metabolic pathways provide suitable targets for new antimycotics, we do not limit these investigations to a single fungal species. Comparisons of different species frequently show that a variety of different solutions has evolved to solve a specific metabolic problem.

Key intermediates of metabolic pathways
Key intermediates of metabolic pathways
In vivo imaging of fungal infections

To study virulence, the impact of specific immune components and the investigation of the efficacy of antifungal drug treatments and murine infection models are frequently applied. In conventional studies selected animals are removed from the experiments and analysed at distinct time points, which only provides a snapshot of the infection process. In contrast, in vivo imaging makes it possible to follow the establishment of infection and disease progression in individual animals in temporal and spatial resolution. This allows to identify cryptic sites of infection and enables the visualisation of drug efficacy in spatial resolution and in real-time.

Especially luminescence based imaging systems possess an excellent signal to noise ratio for in vivo investigations. Therefore, we are establishing bioluminescent reporter strains of different fungal species. To generate these strains, we construct synthetic reporter genes that are adapted to the specific target organism and are cloned under the control of strong promoters. Currently, we are focussing on the generation of bioluminescent Aspergilli, Candida species and different Cryptococcus neoformans serotypes.

Visualisation of Candida albicans persistence in the gall bladder. A.) In vivo Imaging. B.) Ex vivo Imaging. C.) Spatial 3D-reconstruction of the origin of the signal from A.
Visualisation of Candida albicans persistence in the gall bladder. A.) In vivo Imaging. B.) Ex vivo Imaging. C.) Spatial 3D-reconstruction of the origin of the signal from A.
Interactions with immune cells

Cells of the innate immune system are of special importance for the recognition, control and elimination of fungal pathogens. Thus, patients with attenuated immune response are of special risk to acquire life-threatening fungal infections. Unfortunately, the impact of the different components of the immune system has only been partially understood. Additionally, the impact of different immune cells may differ from pathogen to pathogen and even closely related fungal species may have developed independent mechanisms to escape the immune response.

We focus our work especially on the differences in immune cell interaction by Aspergillus fumigatus and Aspergillus terreus. We showed that both species interact differently with alveolar macrophages, which form a first line of defence against fungal spores. A. fumigatus inhibits acidification of phagolysosomes and rapidly escapes macrophages by elongating hyphae. In contrast, A. terreus does not inhibit this acidification, but persists within the phagolysosome for days or weeks. This difference is mainly based on the pigments present in the spores that are of different origin in both species. Since A. terreus infections are much more frequently associated with disseminations, we are currently investigating the possibility of using immune cells as a vehicle for dissemination within the host.

Aspergillus fumigatus escape from macrophages
Aspergillus fumigatus escape from macrophages
Secondary metabolites from A. terreus

Aspergillus terreus is known for its ability to produce the primary metabolite itaconic acid and the secondary metabolite lovastatin. While itaconic acid is an interesting intermediate for several chemical processes, lovastatin is used for the reduction of cholesterol levels in patients by inhibiting the HMG-CoA reductase. Interestingly, genome analyses have shown that A. terreus possesses a much higher potential of producing natural products than has been identified so far.

In our studies, we try to activate selected gene clusters from A. terreus. The aim is the identification of novel metabolites and the characterisation of their biologic activities. A special attention is given to the backwards correlation of the biologic activities and the environmental conditions leading to its production. Investigation of gene expression under natural conditions additionally involves the generation of reporter strains that indicate the expression of the cluster under selected conditions. Due to these studies we have recently been able to provide a correlation between the production of the natural product terrein and plant interactions. Plant derived media are strong inducers for terrein production, whereby terrein can harm the surfaces of fruits and suppresses growth of plant seedlings. Thus, terrein production seems to have evolved as an adaptation to life within the rhizospheres. More detailed characterisations and other metabolites are currently under investigation.

Terrein-induced lesions on banana skin surface
Terrein-induced lesions on banana skin surface

Publications

2015

Brock M (2015) Bringing light into the dark site of infection. Cytometry Part A [Epub ahead of print] (Review) Details PubMed

Gressler M, Hortschansky P, Geib E, Brock M (2015) A new high-performance heterologous fungal expression system based on regulatory elements from the Aspergillus terreus terrein gene cluster. Front Microbiol 6, 184. Details PubMed

Gressler M, Meyer F, Heine D, Hortschansky P, Hertweck C, Brock M (2015) Phytotoxin production in Aspergillus terreus is regulated by independent environmental signals. eLife 4, e07861. Details PubMed Open Access PDF

2014

Brunke S, Seider K, Richter ME, Bremer-Streck S, Ramachandra S, Kiehntopf M, Brock M, Hube B (2014) Histidine degradation via an aminotransferase increases the nutritional flexibility of Candida glabrata. Eukaryot Cell 13(6), 758-765. Details PubMed

Jacobsen ID, Lüttich A, Kurzai O, Hube B, Brock M (2014) In vivo imaging of disseminated murine Candida albicans infection reveals unexpected host sites of fungal persistence during antifungal therapy. J Antimicrob Chemother 69(10), 2785-2796. Details PubMed

Otzen C, Bardl B, Jacobsen ID, Nett M, Brock M (2014) Candida albicans utilizes a modified β-oxidation pathway for the degradation of toxic propionyl-CoA. J Biol Chem 289(12), 8151-8169. Details PubMed

Papon N, Courdavault V, Lanoue A, Clastre M, Brock M (2014) Illuminating Fungal Infections with Bioluminescence PLOS Pathog. 10(7), e1004179. (Review) Details PubMed

Ramachandra S, Linde J, Brock M, Guthke R, Hube B, Brunke S (2014) Regulatory networks controlling nitrogen sensing and uptake in Candida albicans. PLOS One 9(3), e92734. Details PubMed Open Access

Zaehle C, Gressler M, Shelest E, Geib E, Hertweck C, Brock M (2014) Terrein Biosynthesis in Aspergillus terreus and Its Impact on Phytotoxicity. Chem Biol 21(6), 719-731. Details PubMed

2013

Fazius F, Zaehle C, Brock M (2013) Lysine biosynthesis in microbes: relevance as drug target and prospects for β-lactam antibiotics production. Appl Microbiol Biotechnol 97(9), 3763-3772. Details PubMed

Galiger C, Brock M, Jouvion G, Savers A, Parlato M, Ibrahim-Granet O (2013) Assessment of efficacy of antifungals against Aspergillus fumigatus: value of real-time bioluminescence imaging. Antimicrob Agents Chemother 57(7), 3046-3059. Details PubMed

Otzen C, Müller S, Jacobsen ID, Brock M (2013) Phylogenetic and phenotypic characterisation of the 3-ketoacyl-CoA thiolase gene family from the opportunistic human pathogenic fungus Candida albicans. FEMS Yeast Res 13(6), 553-564. Details PubMed

previously

Brock M (2012) Application of bioluminescence imaging for in vivo monitoring of fungal infections. Int J Microbiol 2012, 956794-956794. Details PubMed

Citiulo F, Jacobsen ID, Miramón P, Schild L, Brunke S, Zipfel PF, Brock M, Hube B, Wilson D (2012) Candida albicans scavenges host zinc via Pra1 during endothelial invasion. PLOS Pathog 8(6), e1002777. Details PubMed Open Access

Fazius F, Shelest E, Gebhardt P, Brock M (2012) The fungal α-aminoadipate pathway for lysine biosynthesis requires two enzymes of the aconitase family for the isomerization of homocitrate to homoisocitrate. Mol Microbiol 86(6), 1508-1530. Details PubMed

Fekkar A, Pionneau C, Brossas JY, Marinach-Patrice C, Snounou G, Brock M, Ibrahim-Granet O, Mazier D (2012) DIGE enables the detection of a putative serum biomarker of fungal origin in a mouse model of invasive aspergillosis. J Proteomics 75(9), 2536-2549. Details PubMed

Jouvion G, Brock M, Droin-Bergère S, Ibrahim-Granet O (2012) Duality of liver and kidney lesions after systemic infection of immunosuppressed and immunocompetent mice with Aspergillus fumigatus. Virulence 3(1), 43-50. Details PubMed

Linde J, Hortschansky P, Fazius E, Brakhage AA, Guthke R, Haas H (2012) Regulatory interactions for iron homeostasis in Aspergillus fumigatus inferred by a Systems Biology approach. BMC Syst Biol 6, 6. Details PubMed

Slesiona S, Gressler M, Mihlan M, Zaehle C, Schaller M, Barz D, Hube B, Jacobsen ID, Brock M (2012) Persistence versus escape: Aspergillus terreus and Aspergillus fumigatus employ different strategies during interactions with macrophages. PLOS One 7(2), e31223-e31223. Details PubMed

Slesiona S, Ibrahim-Granet O, Olias P, Brock M, Jacobsen ID (2012) Murine infection models for Aspergillus terreus pulmonary aspergillosis reveal long-term persistence of conidia and liver degeneration. J Infect Dis 205(8), 1268-1277. Details PubMed

Brock M (2011) Weiterentwicklung des Hühnerei-Infektionsmodells und neue Biolumineszenz-Detektionssysteme liefern genauere Aussagen bei Pilzinfektionen und antifungalen Therapien SIEMENS Forum Aktuell (4), 11-11. Details

Cruz AH, Brock M, Zambuzzi-Carvalho PF, Santos-Silva LK, Troian RF, Góes AM, Soares CM, Pereira M (2011) Phosphorylation is the major mechanism regulating isocitrate lyase activity in Paracoccidioides brasiliensis yeast cells. FEBS J 278(13), 2318-2332. Details PubMed

Fleck CB, Schöbel F, Brock M (2011) Nutrient acquisition by pathogenic fungi: nutrient availability, pathway regulation, and differences in substrate utilization. Int J Med Microbiol 301(5), 400-407. Details PubMed

Gressler M, Zaehle C, Scherlach K, Hertweck C, Brock M (2011) Multifactorial induction of an orphan PKS-NRPS gene cluster in Aspergillus terreus. Chem Biol 18(2), 198-209. Details PubMed

Müller S, Fleck CB, Wilson D, Hummert C, Hube B, Brock M (2011) Gene acquisition, duplication and metabolic specification: the evolution of fungal methylisocitrate lyases. Environ Microbiol 13(6), 1534-1548. Details PubMed

Olias P, Gruber AD, Hafez HM, Lierz M, Slesiona S, Brock M, Jacobsen ID (2011) Molecular epidemiology and virulence assessment of Aspergillus fumigatus isolates from white stork chicks and their environment. Vet Microbiol 148(2-4), 348-355. Details PubMed

Rezola A, de Figueiredo LF, Brock M, Pey J, Podhorski A, Wittmann C, Schuster S, Bockmayr A, Planes FJ (2011) Exploring metabolic pathways in genome-scale networks via generating flux modes. Bioinformatics 27(4), 534-540. Details PubMed

Brunke S, Seider K, Almeida RS, Heyken A, Fleck CB, Brock M, Barz D, Rupp S, Hube B (2010) Candida glabrata tryptophan-based pigment production via the Ehrlich pathway. Mol Microbiol 76(1), 25-47. Details PubMed

Jacobsen ID, Grosse K, Slesiona S, Hube B, Berndt A, Brock M (2010) Embryonated eggs as an alternative infection model to investigate Aspergillus fumigatus virulence. Infect Immun 78(7), 2995-3006. Details PubMed

Schöbel F, Jacobsen ID, Brock M (2010) Evaluation of lysine biosynthesis as an antifungal drug target: biochemical characterization of Aspergillus fumigatus homocitrate synthase and virulence studies. Eukaryot Cell 9(6), 878-893. Details PubMed

Domin N, Wilson D, Brock M (2009) Methylcitrate cycle activation during adaptation of Fusarium solani and Fusarium verticillioides to propionyl-CoA-generating carbon sources. Microbiology 155(Pt 12), 3903-3912. Details PubMed

Ibrahim-Granet O, Dubourdeau M, Latgé JP, Ave P, Huerre M, Brakhage AA, Brock M (2008) Methylcitrate synthase from Aspergillus fumigatus is essential for manifestation of invasive aspergillosis. Cell Microbiol 10(1), 134-148. Details PubMed

Behnsen J, Narang P, Hasenberg M, Gunzer F, Bilitewski U, Klippel N, Rohde M, Brock M, Brakhage AA, Gunzer M (2007) Environmental dimensionality controls the interaction of phagocytes with the pathogenic fungi Aspergillus fumigatus and Candida albicans. PLOS Pathog 3(2), e13. Details PubMed

Gebhardt P, Dornberger K, Gollmick FA, Gräfe U, Härtl A, Görls H, Schlegel B, Hertweck C (2007) Quercinol, an anti-inflammatory chromene from the wood-rotting fungus Daedalea quercina (Oak Mazegill). Bioorg Med Chem Lett 17(9), 2558-2560. Details PubMed

Schöbel F, Ibrahim-Granet O, Avé P, Latgé JP, Brakhage AA, Brock M (2007) Aspergillus fumigatus does not require fatty acid metabolism via isocitrate lyase for development of invasive aspergillosis. Infect Immun 75(3), 1237-1244. Details PubMed

Ebel F, Schwienbacher M, Beyer J, Heesemann J, Brakhage AA, Brock M (2006) Analysis of the regulation, expression, and localisation of the isocitrate lyase from Aspergillus fumigatus, a potential target for antifungal drug development. Fungal Genet Biol 43(7), 476-489. Details PubMed

Sugareva V, Härtl A, Brock M, Hübner K, Rohde M, Heinekamp T, Brakhage AA (2006) Characterisation of the laccase-encoding gene abr2 of the dihydroxynaphthalene-like melanin gene cluster of Aspergillus fumigatus. Arch Microbiol 186(5), 345-355. Details PubMed