TCRsafe® analysis of a Runx1 knockout mouse model provides evidence for a role of RUNX1 as a recombinase cofactor for TCRbeta rearrangements and pathological deletions

Seitz et al. Sci Rep. 2020;10(1):10024. <Free article>

In this work, a Runx1 knockout mouse model was analyzed with quantitative TCRsafe® technology, which was optimized employing a reference template. This ensured precise measurement of changes in the murine T-cell receptor (TCR) repertoire as a consequence of Runx1 functionality. It could be demonstrated that Runx1 binds to the initiation site of TCRbeta rearrangement and its homozygous inactivation induces severe structural changes of the rearranged TCRbeta gene, whereas heterozygous inactivation had almost no impact. Furthermore, published sequence data was reanalyzed from recurrent deletion borders of ALL patients carrying an ETV6-RUNX1 translocation, demonstrating that RUNX1 motifs were significantly overrepresented at these deletion ends. Additionally, it was shown that a RUNX1 motif overlapping with a cryptic recombination signal sequence at a recurrent breakpoint in the CDKN2A/B locus was functionally relevant. As TCRbeta rearrangement can be seen as a physiological deletion process, our data collectively imply a role of RUNX1 as recombinase cofactor for both physiological and aberrant deletions.




ZIM-Project TCR-specific Antigen Identification - TAgID

TAgID is an HSD project funded by the Central Innovation Programme for small and medium-sized enterprises (SMEs; Zentrales Innovationsprogramm Mittelstand, ZIM) in partnership with the Technical University of Berlin. The collaborative activities aim at developing an innovative approach for the identification of peptide/MHC-complexes recognized by tumor-specific T cell receptors (TCRs) (https://www.zim.de/ZIM/Navigation/DE/Meta/Englisch/englisch.html, Grant number: 16KN073238.)




Patent granted for the identification of tumor-specific T-cell receptors

The granted patent (EP3180433B1) “Method for providing tumor-specific T cells” encompasses the quantitative TCRsafe® high throughput T-cell receptor (TCR) repertoire analysis of tumor infiltrating T-cells compared to T-cells infiltrating adjacent corresponding normal tissue (and/or present in peripheral blood) and an in-house developed software package, which enables the identification of the tumor-specific TCR repertoire for each individual patient. Complete decoding of paired alpha and beta chains of tumor-specific TCRs facilitates synthesis and cloning of TCRs that can be applied to genetic engineering of patients’ autologous T cells. These engineered T cells are used as tumor-specific probes for the discovery of tumor antigens. They can also be used for adoptive T cell therapy (see https://www.therycell.de).




Dr. Volker Lennerz joins the HSDiagnomics advisory board

Volker Lennerz is a senior scientist in the group of Prof. Wölfel at the University Clinics Mainz (Medizinische Klinik III) and has published pioneering studies about techniques for unraveling tumor associated neo-antigens in melanoma (https://www.unimedizin-mainz.de/3-med/arbeitsgruppen/ag-woelfel/ag-woelfel/publikationen.html). He contributed to the identification and molecular characterization of numerous tumor antigens in a variety of patient models using different technology platforms, e.g. cDNA expression cloning. V.L. will give important impact to HSDs novel approach to identify and use tumor-specific T-cell receptors to discover novel and public tumor neo-antigens in lung cancer and other cancer types.




Patent granted for TCRsafe® analysis

The granted patent “Methods and primer sets for high throughput PCR sequencing” (EP2746405B1) describes a new method to prevent carry-over contaminations in two-step PCR next generation sequencing (NGS) library preparations and for an accurate analysis of T-cell receptor repertoires (TCRsafe®). The technology featuring an intrinsic contamination protection system effectively prevents cross-sample-contaminations, which is of great advantage considering the distinct sensitivity of current NGS technologies. This technology is a prerequisite for the identification of dominant, tumor-specific T cells and their TCRs and applied to lung cancer, breast cancer, ovarian cancer, prostate cancer, melanoma specimens, etc. The TCRsafe® method was described in Seitz et al. Nucleic Acids Res. 2015. <Free article>




Prof. Dr. Armin Gerbitz joins the HSDiagnomics advisory board

Armin Gerbitz is professor for “Hematology and Cellular Therapy” at the Dep. of Medicine 5, Hematology/Oncology, Hematopoietic Stem Cell Transplantation, University Hospital of Erlangen. For more than 15 years his main research focus lies in allogeneic hematopoietic stem cell transplantation, diagnostics and therapies of malignant lymphomas and leukemia, and therapy related problems like Graft-versus-Host-Disease (GvHD) and reactivation of latent herpes viruses. He is an expert in human and murine immune cell biology and has long-standing expertise in murine model systems. Within HSDiagnomis AG is a senior advisor for all aspects of T-cell profiling, related technologies, and upcoming applications in T-cell based treatments and diagnostics, e.g. novel tumor therapies.




HSDiagnomics launches TCRsafe® T-cell profiling service

As of 10/31/2014 HSDiagnomics starts offering its TCRsafe® T-cell profiling service to the research community. TCRsafe® is the only T-cell profiling service world-wide, which provides rigorous deep-sequencing of the complete TCR-beta repertoire in combination with an inbuilt highly effective contamination protection system to eliminate even spurious alien DNA from input samples. The mission of HSDiagnomics will be to provide customers from clinical or research labs with a complete and very fast T-cell profiling service down to the single-cell level.




HSDiagnomics wins ProFIT grant provided by Investitionsbank Berlin (IBB)

We are very grateful to the IBB, the city of Berlin, and the European Union, that the concept of a novel method for T-cell profiling for research and diagnostic applications was elected for a 2 years period funding, which enabled HSDiagnomics to establish a molecular biology lab in Berlin-Steglitz and to develop and refine methods for deep TCRbeta-profiling. The ProFIT program is financed by the city of Berlin as well as by the European Union (EFRE, Europäischer Fonds für regionale Entwicklung).




HSDiagnomics enters partnership with Charite/Berlin

In the context of the ProFIT program (see above) HSDiagnomics and Charite Dept. of Molecular Pathology Prof. Michael Hummel, agreed on a close partnership for development of deep T-cell profiling technologies and pilot applications in a series of research projects, covering areas like lymphomas, GvHD, coeliac disease. A special focus will be on practical tests of HSDiagnomics double-contamination-protection system. (TCRsafe®).