... in times of the Corona crisis
Therapeutic Gene Regulation
The current Covid-19 pandemic is an impressive illustration of how differently individual people respond to a pathogen. Some people do not develop any symptoms at all – others die from it. Against some pathogens we develop lifelong immunity, against others we do not. The reasons for these differences are largely unknown. In order to study the underlying reasons, we need analytical methods for the precise and systematic determination of human immune reactions. The DRFZ has developed unique methods to identify the cells of the human immune system that participate in an immune reaction and to isolate them from tissue and blood.
Mashreghi’s research group is specialized in studying these cells individually in order to determine (1) which genes they express, suggesting their role in the immune response, how they were activated and whether they represent immune memory cells. In addition, the analysis helps to identify (2) the antigen receptors expressed on the adaptive immune cells. This reveals the people who have these receptors, which antigens these receptors recognize, how the immune cells are related to each other and how they contribute to immunity. This expertise is technologically based on the isolation of reactive lymphocytes, the use of oligonucleotide-coupled antibodies for labelling of surface proteins (CITE-Seq), the generation of 10X Genomics based single cell cDNA libraries (10X Genomics 5`Next GEM) followed by NGS sequencing (Illumina), the analysis of the antigen receptor repertoire (single-cell V(D)J sequencing) of B and T lymphocytes at the single-cell level in the context of their global gene and surface protein signature, including the primary and secondary biocomputational analysis of the data (bioinformatics platform).
The expertise of this research group offers a unique opportunity to study immune responses, in particular currently, in collaboration with virological and clinical research groups, to investigate the heterogeneity of human immune responses to SARS-CoV2 (Covid-2).
About the lab
We aim to identify and characterize cells that trigger and drive the pathogenesis of rheumatic diseases. We apply state-of-the-art methods (particularly Single-Cell-Sequencing approaches) to determine which genes and regulatory ribonucleic acids (RNAs) are selectively switched on in “disease causing” pathogenic cells versus healthy cells. The identified genes and regulatory RNAs are further functionally characterized in order to elucidate whether they influence the disease directly or indirectly. Genes and regulatory RNAs which keep the pathogenic cells alive would be ideal targets for new therapies. Their suitability as novel therapeutic targets is tested in preclinical models for rheumatic diseases.
All the identified regulatory RNAs and many of the identified genes that are selectively expressed by cells which are involved in the pathogenesis of rheumatic diseases, encode for proteins that act inside these cells, making them inaccessible to modern biologics. We are therefore developing short nucleic acids (oligonucleotides) that inhibit the RNAs that translate information from these genes into proteins. Such oligonucleotides are small inhibitory RNAs (siRNAs) that directly inhibit gene expression, and antagomirs that inhibit regulatory microRNAs and thus indirectly influence gene expression. When these oligonucleotides are coupled to cholesterol, they easily enter all cells, but only affect gene expression in the cells in which a particular gene or regulatory RNA is switched on in, making them therefore, very selective. We want to utilize them for therapeutic manipulation of chondrocytes in patients with osteoarthritis (OA). In OA, chondrocytes fail to build cartilage tissue or even destroy the existing extracellular matrix by secreting matrix-degrading enzymes. In cooperation with the Löhning group, we are determining target genes in order to reactivate the cartilage production of degenerated chondrocytes or to turn precursor cells into active chondrocytes.
Therapeutic oligonucleotides also have great potential for the treatment of chronic inflammatory rheumatic diseases. In an animal model, we were able to show that antagomirs against the microRNA-148a specifically deplete the disease-causing T-helper (Th) lymphocytes and significantly attenuate chronic inflammation due to the dependence on miR-148a for their survival. Protective memory Th lymphocytes generated by a vaccine-like immunization were not affected because they do not express miR-148a. We have thus provided the fundamental proof that therapeutic oligonucleotides can act selectively and efficiently in the organism without significant undesirable side effects.
Dr. rer. nat. Mir-Farzin Mashreghi
Dr. rer. nat. Gitta Anne Heinz
Dr. rer nat. Patrick Maschmeyer
Dr. rer. nat. Frederik Heinrich
- PD Dr. med. Tilmann Kallinich, Department of Pediatrics, Charité-Universitätsmedizin Berlin
- Prof. Dr. Marcus Mall, Charité, Department of Pediatrics, Charité-Universitätsmedizin Berlin
- Prof. Dr. Hans-Dieter Volk, BCRT
- Prof. Dr. Petra Reinke, BCRT
- Dr. Antigoni Triantafyllopoulou, DRFZ
- Dr. Szandor Simmons, Charité
- Dr. Tobias Alexander, Department of Rheumatology and Clinical Immunology, Charité-Universitätsmedizin Berlin
- Dr. Klemens Budde, Department of Nephrology, Charité-Universitätsmedizin Berlin
- Prof. Dr. Falk Hiepe, Department of Rheumatology and Clinical Immunology, Charité-Universitätsmedizin Berlin
- Jürgen Wittmann, Molecular Immunology, Universitätsklinikum Erlangen
- PD Dr. Anja Kühl, Institute for Pathology, Charité-Universitätsmedizin Berlin
- Dr. Michael Lohoff, Institute for Medical Mikrobiology Philipps University Marburg
- Dr. Stefan Kaufmann, Max-Planck-Institute for Infektion Biology
- Dr. Fritz Melchers, DRFZ
- Dr. Andreas Hutloff, DRFZ
- Dr. Andrey Kruglov, DRFZ
- Dr. Chiara Romagnani, DRFZ
- Prof. Dr. Andreas Radbruch, DRFZ
- Prof. Dr. Andreas Diefenbach, DRFZ
- Prof. Dr. Max Löhning, DRFZ
- Dr. Bastian Opitz, Department of Infectious Diseases and Pulmonary Medicine, Charité-Universitätsmedizin Berlin
- Prof. Dr. Simon Fillatreau, Institut Necker-enfants-malades (INEM), Paris, France.
- George Goulielmos, Molecular Medicine and Human Genetics Laboratory, University of Crete, Heraklion, Greece.
- Dr. Andreas Krüger, institute for Molecular Medicine, Goethe Universität Frankfurt
- Dr. Dirk Ostareck, Department of Intensive Care, University Hospital Aachen
- Dr. Antje Ostareck-Lederer, Department of Intensive Care University Hospital Aachen
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- Bardua M, Haftmann C, Durek P, Westendorf K, Buttgereit A, Tran CL, McGrath M, Weber M, Lehmann K, Addo RK, Heinz GA, Stittrich AB, Maschmeyer P, Radbruch H, Lohoff M, Chang HD, Radbruch A, Mashreghi MF. MicroRNA-31 Reduces the Motility of Proinflammatory T Helper 1 Lymphocytes. Front Immunol. 2018 Dec 6;9:2813. doi: 10.3389/fimmu.2018.02813. eCollection 2018. PubMed PMID: 30574141; PubMed Central PMCID: PMC6291424.
- Matz M, Heinrich F, Zhang Q, Lorkowski C, Seelow E, Wu K, Lachmann N, Addo RK, Durek P, Mashreghi MF, Budde K. The regulation of interferon type I pathway-related genes RSAD2 and ETV7 specifically indicates antibody-mediated rejection after kidney transplantation. Clin Transplant. 2018 Dec;32(12):e13429 doi: 10.1111/ctr.13429. Epub 2018 Nov 18. PubMed PMID: 30341925.
- Matz M, Heinrich F, Lorkowski C, Wu K, Klotsche J, Zhang Q, Lachmann N, Durek P, Budde K, Mashreghi MF. MicroRNA regulation in blood cells of renal transplanted patients with interstitial fibrosis/tubular atrophy and antibody-mediated rejection. PLoS One. 2018 Aug 13;13(8):e0201925. doi: 10.1371/journal.pone.0201925. eCollection 2018. PubMed PMID: 30102719; PubMed Central PMCID: PMC6089438.
- Hammer Q, Rückert T, Borst EM, Dunst J, Haubner A, Durek P, Heinrich F, Gasparoni G, Babic M, Tomic A, Pietra G, Nienen M, Blau IW, Hofmann J, Na IK, Prinz I, Koenecke C, Hemmati P, Babel N, Arnold R, Walter J, Thurley K, Mashreghi MF, Messerle M, Romagnani C. Peptide-specific recognition of human cytomegalovirus strains controls adaptive natural killer cells. Nat Immunol. 2018 May;19(5):453-463. doi: 10.1038/s41590-018-0082-6. Epub 2018 Apr 9. PubMed PMID:
- Lahmann A, Kuhrau J, Fuhrmann F, Heinrich F, Bauer L, Durek P, Mashreghi MF, Hutloff A. Bach2 Controls T Follicular Helper Cells by Direct Repression of Bcl-6. J Immunol. 2019 Apr 15;202(8):2229-2239. doi: 10.4049/jimmunol.1801400. Epub 2019 Mar 4. PubMed PMID: 30833348.
- Neumann C, Blume J, Roy U, Teh PP, Vasanthakumar A, Beller A, Liao Y, Heinrich F, Arenzana TL, Hackney JA, Eidenschenk C, Gálvez EJC, Stehle C, Heinz GA, Maschmeyer P, Sidwell T, Hu Y, Amsen D, Romagnani C, Chang HD, Kruglov A Mashreghi MF, Shi W, Strowig T, Rutz S, Kallies A, Scheffold A. c-Maf-dependent T(reg) cell control of intestinal T(H)17 cells and IgA establishe host-microbiota homeostasis. Nat Immunol. 2019 Apr;20(4):471-481. doi: 10.1038/s41590-019-0316-2. Epub 2019 Feb 18. PubMed PMID: 30778241.
- Maschmeyer P, Petkau G, Siracusa F, Zimmermann J, Zügel F, Kühl AA, Lehmann K, Schimmelpfennig S, Weber M, Haftmann C, Riedel R, Bardua M, Heinz GA, Tran CL, Hoyer BF, Hiepe F, Herzog S, Wittmann J, Rajewsky N, Melchers FG, Chang HD, Radbruch A, Mashreghi MF. Selective targeting of pro-inflammatory Th1 cells by microRNA-148a-specific antagomirs in vivo. J Autoimmun. 2018 May;89:41-52. Doi 10.1016/j.jaut.2017.11.005. Epub 2017 Dec 1. PubMed PMID: 29183643; PubMed Central PMCID: PMC5916452.
- Bothur E, Raifer H, Haftmann C, Stittrich AB, Brüstle A, Brenner D, Bollig N, Bieringer M, Kang CH, Reinhard K, Camara B, Huber M, Visekruna A, Steinhoff U, Repenning A, Bauer UM, Sexl V, Radbruch A, Sparwasser T, Mashreghi MF, Wah Mak T, Lohoff M. Antigen receptor-mediated depletion of FOXP3 in induced regulatory T-lymphocytes via PTPN2 and FOXO1. Nat Commun. 2015 Oct 13;6:8576. doi: 10.1038/ncomms9576. PubMed PMID: 26815406; PubMed Central PMCID: PMC4633965.
- Matz M, Fabritius K, Lorkowski C, Dürr M, Gaedeke J, Durek P, Grün JR, Goestemeyer A, Bachmann F, Wu K, Rudolph B, Schmidt D, Weber U, Haftmann C Unterwalder N, Lachmann N, Radbruch A, Neumayer HH, Mashreghi MF, Budde K Identification of T Cell-Mediated Vascular Rejection After Kidney Transplantatio by the Combined Measurement of 5 Specific MicroRNAs in Blood. Transplantation. 2016 Apr;100(4):898-907. doi: 10.1097/TP.0000000000000873. PubMed PMID: 26444957.
- Haftmann C, Riedel R, Porstner M, Wittmann J, Chang HD, Radbruch A, Mashreghi MF. Direct uptake of Antagomirs and efficient knockdown of miRNA in primary B and T lymphocytes. J Immunol Methods. 2015 Nov;426:128-33. doi: 1016/j.jim.2015.07.006. Epub 2015 Jul 15. PubMed PMID: 26187895; PubMe Central PMCID: PMC4655414.
- Weber JP, Fuhrmann F, Feist RK, Lahmann A, Al Baz MS, Gentz LJ, Vu Van D, Mages HW, Haftmann C, Riedel R, Grün JR, Schuh W, Kroczek RA, Radbruch A Mashreghi MF, Hutloff A. ICOS maintains the T follicular helper cell phenotype b down-regulating Krüppel-like factor 2. J Exp Med. 2015 Feb 9;212(2):217-33. doi: 10.1084/jem.20141432. Epub 2015 Feb 2. PubMed PMID: 25646266; PubMed Central PMCID: PMC4322049.
- Haftmann C, Stittrich AB, Zimmermann J, Fang Z, Hradilkova K, Bardua M, Westendorf K, Heinz GA, Riedel R, Siede J, Lehmann K, Weinberger EE, Zimmel D, Lauer U, Häupl T, Sieper J, Backhaus M, Neumann C, Hoffmann U, Porstner M, Chen W, Grün JR, Baumgrass R, Matz M, Löhning M, Scheffold A, Wittmann J, Chang HD, Rajewsky N, Jäck HM, Radbruch A, Mashreghi MF. miR-148a is upregulated by Twist1 and T-bet and promotes Th1-cell survival by regulating the proapoptotic gene Bim. Eur J Immunol. 2015 Apr;45(4):1192-205. doi: 10.1002/eji.201444633. Epub 2015 Jan PubMed PMID: 25486906; PubMed Central PMCID: PMC4406154
- Stittrich AB, Haftmann C, Sgouroudis E, Kühl AA, Hegazy AN, Panse I, Riedel R, Flossdorf M, Dong J, Fuhrmann F, Heinz GA, Fang Z, Li N, Bissels U, Hatam F, Jahn A, Hammoud B, Matz M, Schulze FM, Baumgrass R, Bosio A, Mollenkopf HJ, Grün J, Thiel A, Chen W, Höfer T, Loddenkemper C, Löhning M, Chang HD, Rajewsky N, Radbruch A, Mashreghi MF. The microRNA miR-182 is induced by IL-2 and promotes clonal expansion of activated helper T lymphocytes. Nat Immunol. 2010 Nov;11(11):1057-62. doi: 10.1038/ni.1945. Epub 2010 Oct 10. PubMed PMID: 20935646.