Modeling NOTCH1 driven T-cell Acute Lymphoblastic Leukemia in Mice

Agnieszka A Wendorff; Adolfo A Ferrando

doi:10.21769/BioProtoc.3620

Modeling NOTCH1 driven T-cell Acute Lymphoblastic Leukemia in Mice

Bio Protoc. 2020 May 20;10(10):e3620. doi: 10.21769/BioProtoc.3620.

Authors

Agnieszka A Wendorff¹, Adolfo A Ferrando^{1

2

3

4}

Affiliations

¹ Institute for Cancer Genetics, Columbia University, New York, USA.
² Department of Pediatrics, Columbia University Medical Center, New York, USA.
³ Department of Systems Biology, Columbia University, New York, USA.
⁴ Department of Pathology and Cell Biology, Columbia University Medical Center, New York, USA.

Abstract

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological malignancy that arises from transformation of T-cell primed hematopoietic progenitors. Although T-ALL is a heterogenous and molecularly complex disease, more than 65% of T-ALL patients carry activating mutations in the NOTCH1 gene. The majority of T-ALL-associated NOTCH1 mutations either disrupt the negative regulatory region, allowing signal activation in the absence of ligand binding, or result in truncation of the C-terminal PEST domain involved in the termination of NOTCH1 signaling by proteasomal degradation. To date, retroviral transduction models have relied heavily on the overexpression of aggressively truncated variants of NOTCH1 (such as ICN1 or ΔE-NOTCH1), which result in supraphysiological levels of signaling activity and are rarely found in human T-ALL. The current protocol describes the method for mouse bone marrow isolation, hematopoietic stem and progenitor cell (HSC) enrichment, followed by retroviral transduction with an oncogenic mutant form of the NOTCH1 receptor (NOTCH1-L1601P-ΔP) that closely resembles the gain-of-function mutations most commonly found in patient samples. A hallmark of this forced expression of constitutively active NOTCH1 is a transient wave of extrathymic immature T-cell development, which precedes oncogenic transformation to T-ALL. Furthermore, this approach models leukemic transformation and progression in vivo by allowing for crosstalk between leukemia cells and the microenvironment, an aspect unaccounted for in cell-line based in vitro studies. Thus, the HSC transduction and transplantation model more faithfully recapitulates development of the human disease, providing a highly comprehensive and versatile tool for further in vivo and ex vivo functional studies.

Keywords: Bone marrow; Hematopoietic stem cell; In vivo; Leukemia development; Mouse model; NOTCH1 mutations; NOTCH1 signaling; Retrovirus; T-ALL; Transformation; Transplantation.

Abstract

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