Abstract
Acute lymphoblastic leukaemia (ALL) is the most common cancer in children. Although aetiology is unclear, various genetic aberrations, such as chromosomal translocations, are suggested to be initiating events in ALL development and have been retrospectively detected at birth, suggesting these events occur in utero. Further factors, such as epigenetic modifications, are also likely required for disease progression. Epidemiological evidence suggests maternal folate status is likely to influence risk of childhood leukaemia. Folate is known to influence DNA damage, repair and methylation status. Current guidelines recommend pregnant women consume synthetic folic acid (FA) supplements during the first trimester, however, biologically active folate, 5-methytetrahydrofolate (5mTHF), is becoming more popular in pregnancy targeted supplements. Our aim is to investigate the influence of FA and 5mTHF on leukaemia-initiators using organoid models to replicate the bone marrow niche.
Mesenchymal stem cells (MSCs) were used to create 2D and 3D organoids supporting growth of GM12878 B cells in media containing physiologically relevant concentrations of FA or 5mTHF. Cell growth was evaluated through trypan blue exclusion. Intracellular and media folate was measured using LC-MS. Reverse-transcription PCR is used to monitor cell markers and identify leukaemia-initiating events.
When grown in co-culture with MSCs, B cell growth is increased compared to growing alone. A further increase in B cell growth was observed when grown in the presence of 3D spheroids compared to 2D monolayers of MSCs. When grown in folate deficient conditions a reduction in B cell growth compared to standard media is observed for both FA and 5mTHF supplementation over 9 days. A reduction in intracellular folate was observed as early as 2 days following folate depletion.
We have optimised a 3D model of folate in B cells, which will allow us to investigate the induction of leukaemia-initiators. Such knowledge may be useful to influence public health policy for folate guidance during pregnancy.
Mesenchymal stem cells (MSCs) were used to create 2D and 3D organoids supporting growth of GM12878 B cells in media containing physiologically relevant concentrations of FA or 5mTHF. Cell growth was evaluated through trypan blue exclusion. Intracellular and media folate was measured using LC-MS. Reverse-transcription PCR is used to monitor cell markers and identify leukaemia-initiating events.
When grown in co-culture with MSCs, B cell growth is increased compared to growing alone. A further increase in B cell growth was observed when grown in the presence of 3D spheroids compared to 2D monolayers of MSCs. When grown in folate deficient conditions a reduction in B cell growth compared to standard media is observed for both FA and 5mTHF supplementation over 9 days. A reduction in intracellular folate was observed as early as 2 days following folate depletion.
We have optimised a 3D model of folate in B cells, which will allow us to investigate the induction of leukaemia-initiators. Such knowledge may be useful to influence public health policy for folate guidance during pregnancy.
| Original language | English |
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| Publication status | Published - 3 Jul 2025 |
| Event | Northumbria University Applied Sciences Departmental Research Day - Newcastle Duration: 3 Jul 2025 → 3 Jul 2025 |
Conference
| Conference | Northumbria University Applied Sciences Departmental Research Day |
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| Period | 3/07/25 → 3/07/25 |
