Identification and Separation of Normal Hematopoietic Stem Cells and Leukemia Stem Cells from Patients with Acute Myeloid Leukemia

Hoang, Van T.; Hoffmann, Isabel; Borowski, Karina; Zepeda‐Moreno, Abraham; Ran, Dan; Buss, Eike C.; Wuchter, Patrick; Eckstein, Volker; Ho, Anthony D.

doi:10.1007/978-1-62703-508-8_19

Cited by 11 publications

(7 citation statements)

References 27 publications

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“…Flow cytometry. Cells were stained with ALDEFLUOR™ reagent (Stemcell Technologies, Inc.) and propidium iodide (PI; BD Biosciences) as previously described (16). Activated ALDEFLUOR™ reagent was aliquoted and stored at -20˚C according to the manufacturer's instructions (Stemcell Technologies, Inc.).…”

Section: Methodsmentioning

confidence: 99%

N,N‑diethylaminobenzaldehyde targets aldehyde dehydrogenase to eradicate human pancreatic cancer cells

Wang

Zheng

et al. 2020

Exp Ther Med

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Pancreatic cancer is a common cause of worldwide cancer-related mortality with a poor 5-year survival rate. Aldehyde dehydrogenase (ALDH) activity is a possible marker for malignant stem cells in solid organ systems, including the pancreas, and N,N-diethylaminobenzaldehyde (DEAB) is able to inhibit ALDH activity. In the present study, the role of DEAB in the treatment of pancreatic cancer cells and the potential underlying mechanisms were investigated. The ALDH activities of pancreatic cancer cell lines treated with or without DEAB were analyzed by an ALDEFLUOR™ assay. The Cell Counting Kit-8 and colony formation assays, and cell cycle analysis were used to evaluate the viability, colony-forming ability and cell quiescence of cell lines under DEAB treatment, respectively. DEAB and/or gemcitabine-induced cell apoptosis was assessed by flow cytometry. DEAB reduced ALDH activity and inhibited the proliferation, colony-forming ability and cell quiescence of pancreatic cancer cell lines. Compared with respective controls, DEAB alone and the combination of gemcitabine and DEAB significantly decreased cell viability and increased cell apoptosis. Moreover, reverse transcription-PCR and western blotting were used to measure the expressions of B cell lymphoma 2 (Bcl2) associated X protein (Bax) and Bcl2 mRNA and protein. The anti-cancer effect of DEAB was associated with upregulation of Bax expression. Therefore, targeting ALDH with DEAB may be a potential therapeutic choice for pancreatic cancer, demonstrating a synergic effect with gemcitabine.

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Section: Methodsmentioning

confidence: 99%

N,N‑diethylaminobenzaldehyde targets aldehyde dehydrogenase to eradicate human pancreatic cancer cells

Wang

Zheng

et al. 2020

Exp Ther Med

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“…MNC were labeled with Aldefluor reagent (Stem Cell Technologies, Vancouver, BC, Canada), CD2‐PE, CD7‐PE, CD11b‐PE, CD15‐PE, CD19‐PE, CD38‐PE, CD56‐PE, CD34‐APC, CD45‐APC‐H7 and propidium iodide (PI; BD Bioscience, Heidelberg, Germany) as described previously . Cells were analyzed using a FACScan flow cytometry system (BD Bioscience, Heidelberg, Germany) equipped with a Rainbow laser (Cytek Flow Cytometry Products, CA), and sorted with a FACSAria II sorter (BD Bioscience, Heidelberg, Germany).…”

Section: Methodsmentioning

confidence: 99%

The rarity of ALDH ⁺ cells is the key to separation of normal versus leukemia stem cells by ALDH activity in AML patients

et al. 2015

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To understand the precise disease driving mechanisms in acute myeloid leukemia (AML), comparison of patient matched hematopoietic stem cells (HSC) and leukemia stem cells (LSC) is essential. In this analysis, we have examined the value of aldehyde dehydrogenase (ALDH) activity in combination with CD34 expression for the separation of HSC from LSC in 104 patients with de novo AML. The majority of AML patients (80 out of 104) had low percentages of cells with high ALDH activity (ALDH+ cells; <1.9%; ALDH‐rare AML), whereas 24 patients had relatively numerous ALDH+ cells (≥1.9%; ALDH‐numerous AML). In patients with ALDH‐rare AML, normal HSC could be separated by their CD34+ALDH+ phenotype, whereas LSC were exclusively detected among CD34+ALDH− cells. For patients with ALDH‐numerous AML, the CD34+ALDH+ subset consisted mainly of LSC and separation from HSC was not feasible. Functional analyses further showed that ALDH+ cells from ALDH‐numerous AML were quiescent, refractory to ARA‐C treatment and capable of leukemic engraftment in a xenogenic mouse transplantation model. Clinically, resistance to chemotherapy and poor long‐term outcome were also characteristic for patients with ALDH‐numerous AML providing an additional risk‐stratification tool. The difference in spectrum and relevance of ALDH activity in the putative LSC populations demonstrates, in addition to phenotypic and genetic, also functional heterogeneity of leukemic cells and suggests divergent roles for ALDH activity in normal HSC versus LSC. By acknowledging these differences our study provides a new and useful tool for prospective identification of AML cases in which separation of HSC from LSC is possible.

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“…High activity of the cytosolic enzyme aldehyde dehydrogenase (ALDH) has been associated with normal haematopoietic stem and progenitor cells (HSPCs) (Storms et al, 1999;Hess et al, 2004;Pearce et al, 2005;Gentry et al, 2007) as well as cancer stem cells (CSCs) in both solid tumours (Ajani et al, 2015) and haematological malignancies (Gerber et al, 2011;Gerber et al, 2012;Hoang et al, 2013;Wu et al, 2013). Moreover, a high level of ALDH activity has been proposed as a tool for separating LSCs from normal HSPCs (Schuurhuis et al, 2013;Hoang et al, 2015).…”

Section: Research Papermentioning

confidence: 99%

Revisiting CLEC12A as leukaemic stem cell marker in AML: highlighting the necessity of precision diagnostics in patients eligible for targeted therapy

et al. 2018

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Summary Targeted therapy directed against rare disease‐propagating leukaemic stem cells (LSCs) is a promising prospect for improving the outcome of acute myeloid leukaemia (AML) patients. Thus, distinguishing LSCs from normal haematopoietic stem and progenitor cells (HSPCs) is essential. The CLEC12A receptor has been proposed as a specific marker of LSCs, and consequently as an appealing treatment target. To explore the role of CLEC12A in further detail, we investigated whether a sorting strategy based on the activity of aldehyde dehydrogenase and CLEC12A expression could separate residual normal HSPCs from LSCs in bone marrow from 5 AML patients. We demonstrate that this distinction was possible in 2/5 cases, however with evidence of pre‐leukaemic mutations in the CLEC12A‐ stem cells in one case. In contrast, cytogenetic and/or molecular aberrations were detected in both the CLEC12A+/− cell subsets in 3/5 AML cases studied. Furthermore, targeted next generation sequencing (NGS) of the sorted cell subsets revealed a pronounced clonal heterogeneity in the CLEC12A‐ cells suggestive of the leukaemia often originating in this immature cell subset. In conclusion, we provide proof‐of‐concept that precision diagnostics employing targeted cytogenetic/NGS‐based analyses on highly purified cell subsets could be a powerful tool for selecting patients eligible for LSC‐directed therapy.

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Identification and Separation of Normal Hematopoietic Stem Cells and Leukemia Stem Cells from Patients with Acute Myeloid Leukemia

Cited by 11 publications

References 27 publications

N,N‑diethylaminobenzaldehyde targets aldehyde dehydrogenase to eradicate human pancreatic cancer cells

N,N‑diethylaminobenzaldehyde targets aldehyde dehydrogenase to eradicate human pancreatic cancer cells

The rarity of ALDH ⁺ cells is the key to separation of normal versus leukemia stem cells by ALDH activity in AML patients

Revisiting CLEC12A as leukaemic stem cell marker in AML: highlighting the necessity of precision diagnostics in patients eligible for targeted therapy

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Identification and Separation of Normal Hematopoietic Stem Cells and Leukemia Stem Cells from Patients with Acute Myeloid Leukemia

Cited by 11 publications

References 27 publications

N,N‑diethylaminobenzaldehyde targets aldehyde dehydrogenase to eradicate human pancreatic cancer cells

N,N‑diethylaminobenzaldehyde targets aldehyde dehydrogenase to eradicate human pancreatic cancer cells

The rarity of ALDH + cells is the key to separation of normal versus leukemia stem cells by ALDH activity in AML patients

Revisiting CLEC12A as leukaemic stem cell marker in AML: highlighting the necessity of precision diagnostics in patients eligible for targeted therapy

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The rarity of ALDH ⁺ cells is the key to separation of normal versus leukemia stem cells by ALDH activity in AML patients