René Geyeregger Group

René Geyeregger Email: rene.geyeregger(at)ccri.at

Clinical Cell Biology and FACS Core Unit

Research focus: Flow cytometry and GMP-compliance work as a supportive interface between research and therapy.

Team

Diagnostics

Most of our work consists of doing diagnostics and providing cytometric cell analyses and cell sorting services, primarily to the St. Anna Children’s Hospital, but also to other labs at the Children’s Cancer Research Institute (CCRI) as well as to external research institutes (mainly through the FACS Core Unit). Over the past years, the number of diagnostic applications has increased considerably and now also includes analyses that address spherocytosis and evaluate human sperm fertility. Every year, over 40,000 cytometric cell analyses and more than 2,000 cell sorts are carried out, the main purpose of these being to control the donor/recipient type of the different blood cells using FISH or PCR methods in post allo-transplantation patients. Extensive internal quality control mechanisms are in place, which conform to internal standard operating procedures (SOPs). In addition, we participate in the regular Austrian and German quality control trials, organized by ÖQUASTA (Austrian Society for Quality Assurance and Standardization of Medical-Diagnostics) and INSTAND (Society for Promotion of Quality Assurance in Medical Laboratories). Our task here is to measure subtypes of blood leukocytes, stem cells, and residual leukocytes in blood products.

Clinical routine

The second largest area contains clinical routine and concerns the manipulation of all cells or blood products that are administered to patients as part of their treatment regimens. These procedures take place under sterile conditions in our GMP laboratory. After successful validation of the system in 2010, we were able to introduce a new freezing medium for the storage of patient cells. The preparation of MNC for clinical use in extracorporeal photopheresis ("mini ECP") proved extraordinarily successful for treatment of Graft-versus-host disease (GVHD) in very young patients. All these procedures do of course require the certification of the laboratory by the AGES according to the tissue safety act (Gewebesicherheitsgesetz) and medicinal product act (Arzneimittelgesetz); they were first approved in 2010 and recently renewed in July 2017.

Clinical phase I/II study of novel T-cell-based immunotherapy

Viral infections are among the most frequent and life-threatening complications following hematopoietic stem cell transplantation (HSCT), especially in paediatric patients. Therefore, our research focus is mainly on the development of novel T-cell-based immunotherapies for the treatment of virus-associated malignancies in patients after HSCT. Based on our novel and fast method for the manufacture of virus-specific T-cells, we initiated a clinical phase I/II trial VISIT (Virus-Specific ImmunoTherapy), which was approved by the AGES in July 2016 and funded by the ZIT Life Science Call (Wirtschaftsagentur).

Research

Our group aims at developing new tests and methods for routine laboratory or clinical applications. Presently, our research interests are focused on the extension of virus-specific T-cell products against Epstein-Barr-Virus (EBV) and Polyomavirus (BKV). More detailed information on the process of engraftment and immune reconstitution (IR) of patients after HSCT will also help to better identify patients at risk for transplant related side effects. For this reason, we are continuously expanding our 10 color flow cytometric assays to >14 colors in order to additionally measure different subtypes of CD34+ hematopoietic stem- and progenitor cells, T-cells, B-cells, NK-cells, monocytes and dendritic cells. FACS Core Unit For more detailed information about this core unit and its services please visit the section FACS Core Unit.

Selected Articles

Hovestadt V, Smith KS, Bihannic L, Filbin MG, Shaw ML, Baumgartner A, DeWitt JC, Groves A, Mayr L, Weisman HR, Richman AR, Shore ME, Goumnerova L, Rosencrance C, Carter RA, Phoenix TN, Hadley JL, Tong Y, Houston J, Ashmun RA, DeCuypere M, Sharma T, Flasch D, Silkov A, Ligon KL, Pomeroy SL, Rivera MN, Rozenblatt-Rosen O, Rusert JM, Wechsler-Reya RJ, Li XN, Peyrl A, Gojo J, Kirchhofer D, Lötsch D, Czech T, Dorfer C, Haberler C, Geyeregger R, Halfmann A, Gawad C, Easton J, Pfister SM, Regev A, Gajjar A, Orr BA, Slavc I, Robinson GW, Bernstein BE, Suvà ML, Northcott PA. Resolving medulloblastoma cellular architecture by single cell genomics. Nature. 2019 Aug;572(7767):74-79. doi: 10.1038/s41586-019-1434-6. Epub 2019 Jul 24.
Neftel C, Laffy J, Filbin MG, Hara T, Shore ME, Rahme GJ, Richman AR, Silverbush D, Shaw ML, Hebert CM, Dewitt J, Gritsch S, Perez EM, Gonzalez Castro LN, Lan X, Druck N, Rodman C, Dionne D, Kaplan A, Bertalan MS, Small J, Pelton K, Becker S, Bonal D, Nguyen QD, Servis RL, Fung JM, Mylvaganam R, Mayr L, Gojo J, Haberler C, Geyeregger R, Czech T, Slavc I, Nahed BV, Curry WT, Carter BS, Wakimoto H, Brastianos PK, Batchelor TT, Stemmer-Rachamimov A, Martinez-Lage M, Frosch MP, Stamenkovic I, Riggi N, Rheinbay E, Monje M, Rozenblatt-Rosen O, Cahill DP, Patel AP, Hunter T, Verma IM, Ligon KL, Louis DN, Regev A, Bernstein BE, Tirosh I, Suvà ML. An integrative model of cellular states, plasticity, and genetics for glioblastoma. Cell. 2019 Aug 8;178(4):835-849.e21. doi: 10.1016/j.cell.2019.06.024. Epub 2019 Jul 18.
Tischer S, Geyeregger R, Kwoczek J, Heim A, Figueiredo C, Blasczyk R, Maecker-Kolhoff B, Eiz-Vesper B. Discovery of immunodominant T-cell epitopes reveals penton protein as a second immunodominant target in human adenovirus infection. J Transl Med. 2016;14(1):286.
Dmytrus J, Matthes-Martin S, Pichler H, Worel N, Geyeregger R, Frank N, Frech C, Fritsch G. Multi-color immune-phenotyping of CD34 subsets reveals unexpected differences between various stem cell sources. Bone Marrow Transplant. 2016;51(8):1093-1100.
Freimuller C, Stemberger J, Artwohl M, Germeroth L, Witt V, Fischer G, Tischer S, Eiz-Vesper B, Knippertz I, Dorrie J, Schaft N, Lion T, Fritsch G, Geyeregger R. Selection of adenovirus-specific and Epstein-Barr virus-specific T cells with major histocompatibility class I streptamers under Good Manufacturing Practice (GMP)-compliant conditions. Cytotherapy. 2015;17(7):989-1007.
Geyeregger R, Freimuller C, Stemberger J, Fischer G, Witt V, Fritsch G. Human AdV-specific T cells: persisting in vitro functionality despite lethal irradiation. Bone Marrow Transplant. 2014;49(7):934-941.
Geyeregger R, Freimuller C, Stemberger J, Artwohl M, Witt V, Lion T, Fischer G, Lawitschka A, Ritter J, Hummel M, Holter W, Fritsch G, Matthes-Martin S. First-in-man clinical results with good manufacturing practice (GMP)-compliant polypeptide-expanded adenovirus-specific T cells after haploidentical hematopoietic stem cell transplantation. J Immunother. 2014;37(4):245-249.