Abstract
Background/aims:
Acute lymphoblastic leukaemia (ALL) is the most common cancer in children. Although aetiology is unclear, various genetic aberrations, such as chromosomal translocations, are considered initiating events in ALL development and have been retrospectively detected at birth, suggesting they occur in utero. Further aberrations, such as epigenetic modifications, are also likely required for disease progression. Epidemiological evidence suggests maternal folate status influences 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. We aim to investigate the influence of FA and 5mTHF on leukaemia-initiators using organoid models to replicate the bone marrow niche.
Methods:
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 levels were measured using LC-MS. Reverse-transcription PCR is used to monitor cell markers and identify leukaemia-initiating events.
Results:
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.
Conclusions:
We have optimised a 3D model of folate status in B cells, allowing us to investigate the induction of leukaemia-initiators. Such knowledge may be useful to influence public health policy for folate guidance during pregnancy.
Acute lymphoblastic leukaemia (ALL) is the most common cancer in children. Although aetiology is unclear, various genetic aberrations, such as chromosomal translocations, are considered initiating events in ALL development and have been retrospectively detected at birth, suggesting they occur in utero. Further aberrations, such as epigenetic modifications, are also likely required for disease progression. Epidemiological evidence suggests maternal folate status influences 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. We aim to investigate the influence of FA and 5mTHF on leukaemia-initiators using organoid models to replicate the bone marrow niche.
Methods:
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 levels were measured using LC-MS. Reverse-transcription PCR is used to monitor cell markers and identify leukaemia-initiating events.
Results:
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.
Conclusions:
We have optimised a 3D model of folate status in B cells, allowing 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 |
|---|---|
| Publication status | Published - 8 Sept 2025 |
| Event | 13th World Congress of the International Society for Developmental Origins of Health and Disease (DOHaD) - Buenos Aires, Argentina Duration: 7 Sept 2025 → 10 Sept 2025 Conference number: 13 https://www.dohad2025.com.ar/ |
Conference
| Conference | 13th World Congress of the International Society for Developmental Origins of Health and Disease (DOHaD) |
|---|---|
| Abbreviated title | 13th World Congress DOHaD |
| Country/Territory | Argentina |
| City | Buenos Aires |
| Period | 7/09/25 → 10/09/25 |
| Internet address |
