Platform 5

Advanced Diagnostics

The 'Advanced Diagnostics' platform aims to establish innovative diagnostic concepts at the CTH. To this end, we are developing new diagnostic tools that bridge the gap between basic science, clinical research, routine diagnostics and patient care. The focus is on applications enabling the translation of research into clinical practice (bench-to-bedside) as well as the feedback of clinical observations into basic research (bedside-to-bench). Thus, this platform serves as a central link between basic research and patient-oriented research and is firmly embedded in the CTH as an integrated centre for clinical and research.

Platform project

Further projects using the platform's expertise

• Prof. Danckwardt: Genomics in Thrombosis and Hemostasis (GinTH) ISTH SSC
• Prof Pierre Morange, Prof. David Alexandre Trégouët, Prof. Danckwardt: non-coding determinants in T&H
• A. Adenäuer, Prof. Schmaier (Case Western Reserve University and UH Cleveland Medical Center), Prof. Lämmle, Prof. Rossmann: Molecular characterisation of historical samples with defects in the kallikrein kininogen system
• Dr. Häuser, Dr. Falter, Dr. Mittler, Dr. Radsak, Dr. Sasca, Prof. Rossmann: Development of ddPCR-based methods for the sensitive quantification of chimerism after bone marrow transplantation and solid organ transplantation from gDNA or cfDNA
• Dr. Häuser, A. Adenäuer, Prof. Rossmann: Characterisation of splice variants to complement modern molecular genetic diagnostics
• Dr. Häuser, A. Adenäuer, PD Dr. O. Beck, Prof. Rossmann, Prof. Lämmle: Importance of molecular genetic tests in the diagnosis of spherocytosis
• S. Popp, Dr. Nitsche, Prof. Rossmann: Integration of functional and molecular genetic diagnostics in primary ciliary dyskinesia (PCD)
• Prof. Vanhoorelbeke (Universität Leuven, Belgien), Prof. Lämmle, Dr. Falter, Prof. Rossmann: Significance of the ADAMTS13 conformation for the pathogenesis of TTP
• Prof. Jurk, A. Adenäuer, Prof. Rossmann: Genetic causes of platelet dysfunction and thrombocythaemia
• Prof. Jurk, Prof. Faber, Prof. Russo, Prof. Rossmann: Patient registry for congenital platelet disorders
• Prof. Lämmle, Dr. v. Auer, Dr. Falter, Prof. Rossmann: Genetic causes and modifiers of TTP (thrombotic thrombocytopenic purpura)
• Prof. Danckwardt, A. Adenäuer, Dr. Häuser, Prof. Rossmann: VWD (von Willebrand disease): Genotype/phenotype ratio
• Dr. Sibbing, Prof. Gori, Prof. Danckwardt, Dr. Wenzel: TROPICAL-ACS (Testing RespOnsiveness To Platelet Inhibition On Chronic AntiplateLet treatment For Acute Coronary Syndromes); contribution of polymorphisms to vascular and platelet function
• Prof. Gori, Prof. Danckwardt: Prevention of in-stent thrombosis
• Dr. Schmidgen, Prof. Gori, Prof. Ruf, Prof. Steinbrink, Prof. Danckwardt: Significance of signalling pathways in cutaneous inflammation
• Dr. Kleis-Fischer, Prof. Grabbe, Prof. Danckwardt: Influence of genetics on wound healing
• This platform is part of the 'Shared Expertise' of the DZHK ('Plattformlobor CTH' - shared expertise SE076)

Particular attention is paid to the molecular analysis of genes that are important for thrombosis and haemostasis. The core of the platform is the sequencing of selected genes using next-generation sequencing (NGS). This gene panel comprises 92 genes with known diagnostic relevance for thrombosis and haemostasis as well as around 300 other genes that are associated with these processes and are of scientific interest to the members of the CTH. The NGS panel is flexible and modular, so that it can be expanded with additional modules if required. This enables customisation to the specific requirements of future cooperation partners, which can be supplemented by additional methods such as whole exome sequencing, Sanger sequencing, pyrosequencing, MLPA (detection of large deletions and insertions), fragment analysis using capillary electrophoresis (for repeat or homopolymer regions), ddPCR (for sensitive allele quantification or quantitative expression analysis) and splice investigations (with and without NMD inhibition; readout: ddPCR, nanopore sequencing).

A further focus of this platform is the analysis of disorders in non-coding sequence sections that predispose to the occurrence of thrombosis or bleeding. This is of particular interest because a proportion of thrombosis and hemostasis disorders do not show significant changes in coding regions. In particular, we investigate the dysregulation of hemostatic components by non-coding RNAs (such as miRNAs, lncRNAs) or RNA-binding proteins. For analysis, we develop and use high-throughput interactome analyzes (miTRAP, (crosslinking) immunoprecipitation (CLIP)) and high-resolution transcriptome analyzes (TRENDseq). With these activities, the platform is internationally anchored in the ISTH SSC Subcommittee on Genomics in Thrombosis and Hemostasis.

To initiate a cooperative project please contact Prof. Heidi Rossmann (Tel. 17-7297), A. Adenäuer (Tel. 17-6904) or Prof. Danckwardt (Tel. 17-6041 or 17-2632)

Scientific Platform Coordiantors

References

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• De Waele L, Sakai K, Mancini I, Sinkovits G, Falter T, Inoue T, Agosti P, Rossmann H, Von Auer C, Tersteeg C, De Meyer SF, Joly BS, Veyradier A, Coppo P, Fijnheer R, Peyvandi F, Prohászka Z, Lämmle B, Vanhoorelbeke K. Open ADAMTS-13 conformation index predicts earlier relapse in immune-mediated thrombotic thrombocytopenic purpura. J Thromb Haemost. 2024 Feb;22(2):493-502.
• Falter T, Rossmann H, de Waele L, Dekimpe C, von Auer C, Müller-Calleja N, Häuser F, Degreif A, Marandiuc D, Messmer X, Sprinzl M, Lackner KJ, Jurk K, Vanhoorelbeke K, Lämmle B. A novel von Willebrand factor multimer ratio as marker of disease activity in thrombotic thrombocytopenic purpura. Blood Adv. 2023 Sep 12;7(17):5091-5102.
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• Ma J, Klemm J, Gerardo-Ramírez M, Frappart L, Castven D, Becker D, Zoch A, Parent R, Bartosch B, Minnich K, Giovannini M, Danckwardt S, Hartmann N, Morrison H, Herrlich P, Marquardt JU, Hartmann M. Cluster of differentiation 44 promotes osteosarcoma progression in mice lacking the tumor suppressor Merlin. Int J Cancer. 2020 Nov 1;147(9):2564-2577.
• Adenaeuer A, Ezigbo ED, Fawzy Nazir H, Barco S, Trinchero A, Laubert-Reh D, Strauch K, Wild PS, Lackner KJ, Lämmle B, Rossmann H. c.451dupT in KLKB1 is common in Nigerians, confirming a higher prevalence of severe prekallikrein deficiency in Africans compared to Europeans. J Thromb Haemost. 2021 Jan;19(1):147-152.
• Roose E, Schelpe AS, Tellier E, Sinkovits G, Joly BS, Dekimpe C, Kaplanski G, Le Besnerais M, Mancini I, Falter T, Von Auer C, Feys HB, Reti M, Rossmann H, Vandenbulcke A, Pareyn I, Voorberg J, Greinacher A, Benhamou Y, Deckmyn H, Fijnheer R, Prohászka Z, Peyvandi F, Lämmle B, Coppo P, De Meyer SF, Veyradier A, Vanhoorelbeke K. Open ADAMTS13, induced by antibodies, is a biomarker for subclinical immune-mediated thrombotic thrombocytopenic purpura. Blood. 2020 Jul 16;136(3):353-361.
• Barco S, Sollfrank S, Trinchero A, Adenaeuer A, Abolghasemi H, Conti L, Häuser F, Kremer Hovinga JA, Lackner KJ, Loewecke F, Miloni E, Vazifeh Shiran N, Tomao L, Wuillemin WA, Zieger B, Lämmle B, Rossmann H. Severe plasma prekallikrein deficiency: Clinical characteristics, novel KLKB1 mutations, and estimated prevalence. J Thromb Haemost. 2020 Jul;18(7):1598-1617.
• Schaupp L, Muth S, Rogell L, Kofoed-Branzk M, Melchior F, Lienenklaus S, Ganal-Vonarburg SC, Klein M, Guendel F, Hain T, Schütze K, Grundmann U, Schmitt V, Dorsch M, Spanier J, Larsen PK, Schwanz T, Jäckel S, Reinhardt C, Bopp T, Danckwardt S, Mahnke K, Heinz GA, Mashreghi MF, Durek P, Kalinke U, Kretz O, Huber TB, Weiss S, Wilhelm C, Macpherson AJ, Schild H, Diefenbach A, Probst HC. Microbiota-Induced Type I Interferons Instruct a Poised Basal State of Dendritic Cells. Cell. 2020 May 28;181(5):1080-1096.e19.
• Ogorodnikov A, Levin M, Tattikota S, Tokalov S, Hoque M, Scherzinger D, Marini F, Poetsch A, Binder H, Macher-Göppinger S, Probst HC, Tian B, Schaefer M, Lackner KJ, Westermann F, Danckwardt S. Transcriptome 3'end organization by PCF11 links alternative polyadenylation to formation and neuronal differentiation of neuroblastoma. Nat Commun. 2018 Dec 14;9(1):5331.
• Nourse J, Braun J, Lackner K, Hüttelmaier S, Danckwardt S. Large-scale identification of functional microRNA targeting reveals cooperative regulation of the hemostatic system. J Thromb Haemost. 2018 Nov;16(11):2233-2245.
• Kargapolova Y, Levin M, Lackner K, Danckwardt S. sCLIP-an integrated platform to study RNA-protein interactomes in biomedical research: identification of CSTF2tau in alternative processing of small nuclear RNAs. Nucleic Acids Res. 2017 Jun 2;45(10):6074-6086.