Modeling Blast Crisis Using Mutagenized Chronic Myeloid Leukemia-Derived Induced Pluripotent Stem Cells (iPSCs)

Abstract

Purpose: To model CML progression in vitro and generate a blast crisis (BC-CML) model in vitro in order to identify new targets. Methods: Three different CML-derived iPSC lines were mutagenized with the alkylating agent ENU on a daily basis for 60 days. Cells were analyzed at D12 of hematopoietic differentiation for their phenotype, clonogenicity, and transcriptomic profile. Single-cell RNA-Seq analysis has been performed at three different time points during hematopoietic differentiation in ENU-treated and untreated cells. Results: One of the CML-iPSCs, compared to its non-mutagenized counterpart, generated myeloid blasts after hematopoietic differentiation, exhibiting monoblastic patterns and expression of cMPO, CD45, CD34, CD33, and CD13. Single-cell transcriptomics revealed a delay of differentiation in the mutated condition as compared to the control with increased levels of MSX1 (mesodermal marker) and a decrease in CD45 and CD41. Bulk transcriptomics analyzed along with the GSE4170 GEO dataset reveal a significant overlap between ENU-treated cells and primary BC cells. Among overexpressed genes, CD25 was identified, and its relevance was confirmed in a cohort of CML patients. Conclusions: iPSCs are a valuable tool to model CML progression and to identify new targets. Here, we show the relevance of CD25 identified in the iPSC model as a marker of CML progression.

Chimeric antigen-receptor (CAR) engineered natural killer cells in a chronic myeloid leukemia (CML) blast crisis model

“…Developing new therapeutic tools against AP/BP-CML is therefore an unmet medical need in this life-threatening step of CML. We and others have shown previously the involvement of CD25 as a biomarker detected in primary BP-CML ( 14 , 20 ) as well as its expression in in vitro BP-CML models ( 21 ). These findings prompted us to design CAR-NK to target CD25 as an off—the—shelf immunotherapy for CML blast crisis.…”

“…We have previously shown that CD25 is overexpressed in BP-CML model generated from patient-specific induced pluripotent stem cells (iPSC) and confirmed the increase of its expression during CML progression ( 21 ). These findings prompted us to evaluate the possibility of targeting this receptor using chimeric antigen receptor (CAR)-based therapy in the setting of CML.…”

“…Here we show the experimental development of a CAR-NK therapy strategy against the IL2RA/CD25 which as described above has been previously reported as being overexpressed, in advanced CML ( 21 ). This strategy is based on the use of a single chain variable fragment (scFv) of the clinically approved monoclonal humanized antibody, Basiliximab recognizing specifically CD25 and targeting K562 cells that we have genetically engineered to overexpress CD25.…”

Design, Synthesis, and Cytotoxic Activities of Indole and Indazole‐Based Stilbenes

“…[1] Chronic myeloid leukemia (CML) is a myeloproliferative disease that starts in hematopoietic systems and migrates to other organs of the body. [2] Breast cancer is the most prevalent tumor affecting women and has become the most commonly diagnosed cancer accounting for 11.7 % of total new cases in 2020. [3] Within different subtypes of breast cancer, triple negative breast cancer (TNBC) is the most aggressive and deadly subtype due to its aggressive biology and poor prognosis.…”

“…Cancer remains a great social burden, causes serious threat to human health throughout the world [1] . Chronic myeloid leukemia (CML) is a myeloproliferative disease that starts in hematopoietic systems and migrates to other organs of the body [2] . Breast cancer is the most prevalent tumor affecting women and has become the most commonly diagnosed cancer accounting for 11.7 % of total new cases in 2020 [3] .…”