Kinase signaling fuels growth of B-cell precursor acute lymphoblastic leukemia (BCP-ALL). Yet its role in leukemia initiation is unclear and has not been shown in primary human hematopoietic cells. We previously described activating mutations in interleukin-7 receptor alpha (IL7RA) in poor-prognosis “ph-like” BCP-ALL. Here we show that expression of activated mutant IL7RA in human CD34+ hematopoietic stem and progenitor cells induces a preleukemic state in transplanted immunodeficient NOD/LtSz-scid IL2Rγnull mice, characterized by persistence of self-renewing Pro-B cells with non-productive V(D)J gene rearrangements. Preleukemic CD34+CD10highCD19+ cells evolve into BCP-ALL with spontaneously acquired Cyclin Dependent Kinase Inhibitor 2 A (CDKN2A) deletions, as commonly observed in primary human BCP-ALL. CRISPR mediated gene silencing of CDKN2A in primary human CD34+ cells transduced with activated IL7RA results in robust development of BCP-ALLs in-vivo. Thus, we demonstrate that constitutive activation of IL7RA can initiate preleukemia in primary human hematopoietic progenitors and cooperates with CDKN2A silencing in progression into BCP-ALL.
Bibliographical noteFunding Information:
We thank Michael Gershovits and Avital Sarusi-Portuguez from the Mantoux bioinformatics institute of the Nancy and Stephen Grand Israel National Center for Personalized Medicine, Weizmann institute of science, for RNAseq analysis service, Dvir Dahary for help with WGS data analysis and Idit Shiff from the Genomic Applications Laboratory, the Core Research Facility, Faculty of Medicine—Ein Kerem, the Hebrew University of Jerusalem, Israel, for 10X RNA sequencing services. We also thank Zhaohui Gu from COH for his assistance regarding PAX5 BCP-ALL subtypes. We are indebted to Nava Gershman, Alla Zozovsky, and Itzhak Ben Moshe for help with xenograft experiments. We thank the Rawlings lab for sharing the Eu-B29 lentiviral construct. Special thanks to Liron Tuval Kochen, Dafna Gaash, Hava Rosenfeld, and Keren Shichrur from the genetic laboratory of Schneider hospital for help in leukemic sample analysis. We thank all past and present members of S.I. research group for fruitful discussions and advice. This work was supported by the Israel Science Foundation (# 1178/12 to S.I.), Children with Cancer (UK) (S.I. and T.E.), Swiss Bridge Foundation (S.I.), WLBH Foundation (S.I.), Waxman Cancer Research Foundation (S.I.), US–Israel Binational Science Foundation, Israeli health ministry ERA-NET program (#CANCER11-FP-127 to S.I.), Hans Neufeld Stiftung, the International Collaboration Grant from the Jacki and Bruce Barron Cancer Research Scholars’ Program, a partnership of the Israel Cancer Research Fund and City of Hope (S.I. grants # 00161), the Nevzlin Genomic Center for Precision Medicine in Schneider Children’s Medical Center of Israel, The European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 813091 (S.I.) and the Israel Childhood Cancer Foundation (S.I.). I.G. was partially supported by Israeli ministry of Immigrant Absorption. This work was performed in partial fulfillment of the requirements for PhD degrees of Ifat Geron, Avigail Rein and Hillary Manriho, Sackler Faculty of Medicine, Tel Aviv University, Israel and of Ifat Geron in the Division of Biological Sciences University of California San Diego, USA.
© 2022, The Author(s).
- Antigens, CD34/genetics
- Base Sequence
- Cell Differentiation/genetics
- Cyclin-Dependent Kinase Inhibitor p16/genetics
- Gene Expression/immunology
- Interleukin-7 Receptor alpha Subunit/genetics
- Mice, Inbred NOD
- Mice, Knockout
- Mice, SCID
- Precursor B-Cell Lymphoblastic Leukemia-Lymphoma/genetics
- Precursor Cells, B-Lymphoid/immunology
- Receptors, Cytokine/genetics
- Signal Transduction/genetics
- Single-Cell Analysis/methods
- Transplantation, Heterologous