Leukemia-initiating HSCs in chronic lymphocytic leukemia reveal clonal leukemogenesis and differential drug sensitivity

Chi-Ling Chiang; Eileen Y Hu; Lingqian Chang; Jadwiga Labanowska; Kevan Zapolnik; Xiaokui Mo; Junfeng Shi; Tzyy-Jye Doong; Arletta Lozanski; Pearlly S Yan; Ralf Bundschuh; Logan A Walker; Daniel Gallego-Perez; Wu Lu; Meixiao Long; Sanggu Kim; Nyla A Heerema; Gerard Lozanski; Jennifer A Woyach; John C Byrd; Ly James Lee; Natarajan Muthusamy

doi:10.1016/j.celrep.2022.111115

Leukemia-initiating HSCs in chronic lymphocytic leukemia reveal clonal leukemogenesis and differential drug sensitivity

Cell Rep. 2022 Jul 19;40(3):111115. doi: 10.1016/j.celrep.2022.111115.

Authors

Chi-Ling Chiang¹, Eileen Y Hu², Lingqian Chang³, Jadwiga Labanowska⁴, Kevan Zapolnik², Xiaokui Mo⁵, Junfeng Shi⁶, Tzyy-Jye Doong⁷, Arletta Lozanski⁷, Pearlly S Yan², Ralf Bundschuh⁸, Logan A Walker⁹, Daniel Gallego-Perez¹⁰, Wu Lu¹¹, Meixiao Long², Sanggu Kim¹², Nyla A Heerema⁴, Gerard Lozanski¹³, Jennifer A Woyach², John C Byrd², Ly James Lee¹⁴, Natarajan Muthusamy¹⁵

Affiliations

¹ Division of Hematology, Department of Internal Medicine, The Ohio State University, Columbus, OH 43210, USA; OSU Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA; Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA.
² Division of Hematology, Department of Internal Medicine, The Ohio State University, Columbus, OH 43210, USA; OSU Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA.
³ Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA; Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA.
⁴ Department of Pathology, The Ohio State University, Columbus, OH 43210, USA.
⁵ Center for Biostatistics, The Ohio State University, Columbus, OH 43210, USA.
⁶ Department of Mechanical and Aerospace Engineering, The Ohio State University, Columbus, OH 43210, USA.
⁷ OSU Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA.
⁸ Division of Hematology, Department of Internal Medicine, The Ohio State University, Columbus, OH 43210, USA; Department of Physics, The Ohio State University, Columbus, OH 43210, USA; Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
⁹ OSU Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA; Department of Physics, The Ohio State University, Columbus, OH 43210, USA.
¹⁰ Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA.
¹¹ Department of Electrical and Computer Engineering, The Ohio State University, Columbus, OH 43210, USA.
¹² Department of Veterinary Biosciences, The Ohio State University, Columbus, OH 43210, USA.
¹³ OSU Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA; Department of Pathology, The Ohio State University, Columbus, OH 43210, USA.
¹⁴ OSU Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA; Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA.
¹⁵ Division of Hematology, Department of Internal Medicine, The Ohio State University, Columbus, OH 43210, USA; OSU Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA; Department of Veterinary Biosciences, The Ohio State University, Columbus, OH 43210, USA. Electronic address: raj.muthusamy@osumc.edu.

PMID: 35858552
DOI: 10.1016/j.celrep.2022.111115

Abstract

The existence of "leukemia-initiating cells" (LICs) in chronic lymphocytic leukemia (CLL) remains controversial due to the difficulty in isolating and identifying the tumor-initiating cells. Here, we demonstrate a microchannel electroporation (MEP) microarray that injects RNA-detecting probes into single live cells, allowing the imaging and characterization of heterogeneous LICs by intracellular RNA expression. Using limited-cell FACS sequencing (LC-FACSeq), we can detect and monitor rare live LICs during leukemogenesis and characterize their differential drug sensitivity. Disease-associated mutation accumulation in developing B lymphoid but not myeloid lineage in CLL patient hematopoietic stem cells (CLL-HSCs), and development of independent clonal CLL-like cells in murine patient-derived xenograft models, suggests the existence of CLL LICs. Furthermore, we identify differential protein ubiquitination and unfolding response signatures in GATA2^high CLL-HSCs that exhibit increased sensitivity to lenalidomide and resistance to fludarabine compared to GATA2^lowCLL-HSCs. These results highlight the existence of therapeutically targetable disease precursors in CLL.

Keywords: CP: Cancer; GATA2; IKZF1; LC-FACSeq; chronic lymphocytic leukemia; fludarabine; lenalidomide; leukemia initiating cells; single-cell analysis; stem cells.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cells, Cultured
Hematopoietic Stem Cells / metabolism
Humans
Leukemia, Lymphocytic, Chronic, B-Cell* / genetics
Leukemia, Lymphocytic, Chronic, B-Cell* / metabolism
Mice
Neoplastic Stem Cells / metabolism
RNA / metabolism

Substances

RNA

Abstract

Publication types

MeSH terms

Substances

Grants and funding