Acute myeloid leukemia (AML), a blood malignancy resulting in abnormal
hematopoiesis, is associated with alterations in the bone marrow environment
(BME). Current treatments for this heterogeneous disease, mainly targeting the
leukemic cells, are largely unsuccessful for the majority of AML subtypes. By better
understanding the mechanisms by which the BME contributes to leukemogenesis, it
may be possible to introduce more effective treatments for AML. Mesenchymal
stromal/stem cells (MSCs) are essential cellular components of the
BME/hematopoietic niche and have been shown to support normal hematopoiesis.
As a critical component, they may have several roles in altering the BME, thus
providing an excellent model for studying the BME in-vitro. Several studies have
characterized AML-derived MSCs (AML-MSCs). However, their exact role in altering
BME remains unclear. Here, we investigated the MSCs' potential role in BME
alteration by investigating the genetic profiles of previously characterized AMLMSCs (n=29) and healthy donor MSCs (HD-MSCs) (n=8). We identified that among
7565 common genes, 21 genes were significantly differentially expressed in AMLMSCs. The CD248 gene was identified among these significantly upregulated genes
in AML/HD-MSCs (n=29). Focusing on AML-MSCs derived from high-risk patients
(HR), CD248 protein was investigated and validated using HR AML-MSCs (n=11)
and HD-MSCs (n=4). Interestingly, it was highly abundant in HR samples at the
intracellular and cell-surface levels. CD248 is an MSC marker and has a biological
significance potentially on their function. To better understand its potential role in
MSC, CD248 was knocked down (KD) in HD-MSCs using siRNA (siCD248-MSCs).
Functional capacity, the ability of HD-MSCs and siCD248-MSCs to differentiate into
cell types that form the BME (adipocytes and osteocytes), and their ability to
promote the growth of HL60 human leukemia cell line were assessed. Posttransfection functional assessments showed that siCD248-MSCs had a reduced
adipogenic but not osteogenic potential via differentiation assays. Quantitative
validation of the adipogenesis pathway by qRT-PCR confirmed the reduction. KD
CD248 increased SIRT2 expression and potentially led to adipogenesis inhibition.
However, co-culture experiments showed no effect of HD-MSCs or siCD248-MSCs on HL60 proliferation. Together these data showed that CD248 is a potential player
in adipogenesis, essential to MSC’s functionality. Thus, it could serve as a
prognostic marker and target for AML therapy.
| Identifer | oai:union.ndltd.org:uottawa.ca/oai:ruor.uottawa.ca:10393/43835 |
| Date | 22 July 2022 |
| Creators | Aldreiwish, Allolo |
| Contributors | Allan, David, Rosu-Myles, Michael |
| Publisher | Université d'Ottawa / University of Ottawa |
| Source Sets | Université d’Ottawa |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Format | application/pdf |
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