Diagnosing recipient- vs. donor-derived posttransplant myelodysplastic neoplasm via targeted single-cell mutational profiling

Jana Ihlow; Livius Penter; Lam Giang Vuong; Philip Bischoff; Benedikt Obermayer; Alexandra Trinks; Olga Blau; Anke Behnke; Thomas Conrad; Markus Morkel; Catherine J Wu; Jörg Westermann; Lars Bullinger; Ann-Christin von Brünneck; Nils Blüthgen; David Horst; Samantha D Praktiknjo

doi:10.1016/j.medj.2024.11.001

Diagnosing recipient- vs. donor-derived posttransplant myelodysplastic neoplasm via targeted single-cell mutational profiling

Med. 2024 Dec 4:S2666-6340(24)00442-2. doi: 10.1016/j.medj.2024.11.001. Online ahead of print.

Authors

Jana Ihlow¹, Livius Penter², Lam Giang Vuong³, Philip Bischoff⁴, Benedikt Obermayer⁵, Alexandra Trinks⁶, Olga Blau⁷, Anke Behnke⁸, Thomas Conrad⁹, Markus Morkel¹⁰, Catherine J Wu¹¹, Jörg Westermann⁷, Lars Bullinger¹², Ann-Christin von Brünneck⁸, Nils Blüthgen¹³, David Horst¹⁴, Samantha D Praktiknjo¹⁵

Affiliations

¹ Institute of Pathology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; Department of Hematology, Oncology and Cancer Immunology, Campus Virchow Clinic, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Berlin, Germany.
² Department of Hematology, Oncology and Cancer Immunology, Campus Virchow Clinic, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Berlin, Germany; Department of Medical Oncology, Dana-Faber Cancer Institute, Boston, MA, USA; Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA, USA; Harvard Medical School, Boston, MA, USA.
³ Department of Hematology, Oncology and Cancer Immunology, Campus Virchow Clinic, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany.
⁴ Institute of Pathology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Berlin, Germany; German Cancer Consortium (DKTK) Partner Site Berlin, German Cancer Research Center (DKFZ), Heidelberg, Germany.
⁵ Core Unit Bioinformatics, Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Berlin, Germany.
⁶ BIH Bioportal Single Cells, Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Berlin, Germany.
⁷ Department of Hematology, Oncology and Cancer Immunology, Campus Virchow Clinic, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; Labor Berlin Charité Vivantes GmbH, Berlin, Germany.
⁸ Institute of Pathology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany.
⁹ Genomics Technology Platform, Berlin Institute of Health at Charité - Universitätsmedizin Berlin and Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany.
¹⁰ Institute of Pathology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; BIH Bioportal Single Cells, Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Berlin, Germany.
¹¹ Department of Medical Oncology, Dana-Faber Cancer Institute, Boston, MA, USA; Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA, USA; Harvard Medical School, Boston, MA, USA.
¹² Department of Hematology, Oncology and Cancer Immunology, Campus Virchow Clinic, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Berlin, Germany; German Cancer Consortium (DKTK) Partner Site Berlin, German Cancer Research Center (DKFZ), Heidelberg, Germany; Labor Berlin Charité Vivantes GmbH, Berlin, Germany.
¹³ Institute of Pathology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; German Cancer Consortium (DKTK) Partner Site Berlin, German Cancer Research Center (DKFZ), Heidelberg, Germany; Institute for Biology, Humboldt-Universität zu Berlin, Berlin, Germany.
¹⁴ Institute of Pathology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; German Cancer Consortium (DKTK) Partner Site Berlin, German Cancer Research Center (DKFZ), Heidelberg, Germany.
¹⁵ Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Berlin, Germany. Electronic address: [email protected].

PMID: 39644889
DOI: 10.1016/j.medj.2024.11.001

Abstract

Background: Distinguishing donor- vs. recipient-derived myelodysplastic neoplasm (MDS) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is challenging and has direct therapeutical implications.

Methods: Here, we took a translational approach that we used in addition to conventional diagnostic techniques to resolve the origin of MDS in a 38-year-old patient with acquired aplastic anemia and evolving MDS after first allo-HSCT. Specifically, we used single-cell transcriptional profiling to differentiate between donor- and recipient-derived bone marrow cells and established a strategy that additionally allows identification of cells carrying the MDS-associated U2AF1^S34Y variant.

Results: The patient exhibited mixed donor chimerism combined with severely reduced erythropoiesis and dysplastic morphology within the granulocytic and megakaryocytic lineage along with the MDS-associated U2AF1^S34Y mutation in the bone marrow. Single-cell transcriptional profiling together with targeted enrichment of the U2AF1^S34Y-specific locus further revealed that, while the immune compartment was mainly populated by donor-derived cells, myelopoiesis was predominantly driven by the recipient. Additionally, concordant with recipient-derived MDS, we found that U2AF1^S34Y-mutated cells were exclusively recipient derived with X but not Y chromosome-specific gene expression.

Conclusion: Our study highlights the clinical potential of integrating high-resolution single-cell techniques to resolve complex cases for personalized treatment decisions.

Funding: The study was funded by intramural resources of the Charité - Universitätsmedizin Berlin and the Berlin Institute of Health.

Keywords: (U2AF1)(S34Y); MDS; Translation to patients; allo-HSCT; aplastic anemia; mixed chimerism; mutational profiling; personalized medicine; precision diagnostics; single-cell; targeted sequencing.