Physiological folate levels constrain nucleotide synthesis and increase dependence on nucleotide salvage
- 1Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA
Abstract
Proliferating cells must acquire nucleotides to support DNA replication, yet how cells meet these nucleotide demands for proliferation under physiological conditions remains understudied. Here, we investigated how physiological nutrient availability shapes nucleotide acquisition strategies in a mouse model of B-cell acute lymphoblastic leukemia (B-ALL). To assess how environmental nutrients impact nucleotide metabolism, we formulated a mouse plasma-like medium (MPM) that reproduces the circulating metabolite composition of plasma from mice with B-ALL and assessed how this influenced nucleotide metabolism relative to standard culture conditions, where nucleotide acquisition has historically been studied. We find that leukemia cells cultured in MPM acquire nucleotides through salvage pathways, and that select nucleotide salvage pathways are required for proliferation under physiological conditions. Of note, this dependency on nucleotide salvage in plasma-like conditions was not caused by precursor metabolite limitation for de novo synthesis. Instead, we found that physiological folate levels are insufficient to support deoxynucleotide triphosphate (dNTP) synthesis for genome replication, leading to DNA replication stress and impaired proliferation when nucleotide salvage is disrupted. Consistently, dietary folate restriction exacerbates the impaired leukemia progression phenotype of nucleotide salvage-deficient B-ALL cells. Together, these findings demonstrate that access to folates is an endogenous limitation for nucleotide synthesis in plasma-like nutrient conditions, increasing the relevance of nucleotide salvage pathways for leukemia progression. More broadly, this work highlights how micronutrient abundance can influence metabolic dependencies and reveals that folate levels shape nucleotide metabolism under physiological conditions.
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BibTeX
@misc{elbashir2026physiological,
title = {{Physiological folate levels constrain nucleotide synthesis and increase dependence on nucleotide salvage}},
author = {R. Elbashir and K. L. Abbott and D. L. Ramesh and A. Ali and A. Shevzov-Zebrun and A. M. Barbeau and M. Wu and A. P. Ward and Y. Gultekin and B. T. Do and S. L. Sivanand and A. Ramos and T. Kunchok and M. Waite and E. H. Rashan and M. Bin Munim and M. Hemann and M. G. Vander Heiden},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.13.750663},
url = {https://www.biorxiv.org/content/10.64898/2026.09.13.750663v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Physiological folate levels constrain nucleotide synthesis and increase dependence on nucleotide salvage
AU - R. Elbashir
AU - K. L. Abbott
AU - D. L. Ramesh
AU - A. Ali
AU - A. Shevzov-Zebrun
AU - A. M. Barbeau
AU - M. Wu
AU - A. P. Ward
AU - Y. Gultekin
AU - B. T. Do
AU - S. L. Sivanand
AU - A. Ramos
AU - T. Kunchok
AU - M. Waite
AU - E. H. Rashan
AU - M. Bin Munim
AU - M. Hemann
AU - M. G. Vander Heiden
PY - 2026
DA - 2026/09/13
PB - bioRxiv
DO - 10.64898/2026.09.13.750663
UR - https://www.biorxiv.org/content/10.64898/2026.09.13.750663v1
AB - Proliferating cells must acquire nucleotides to support DNA replication, yet how cells meet these nucleotide demands for proliferation under physiological conditions remains understudied. Here, we investigated how physiological nutrient availability shapes nucleotide acquisition strategies in a mouse model of B-cell acute lymphoblastic leukemia (B-ALL). To assess how environmental nutrients impact nucleotide metabolism, we formulated a mouse plasma-like medium (MPM) that reproduces the circulating metabolite composition of plasma from mice with B-ALL and assessed how this influenced nucleotide metabolism relative to standard culture conditions, where nucleotide acquisition has historically been studied. We find that leukemia cells cultured in MPM acquire nucleotides through salvage pathways, and that select nucleotide salvage pathways are required for proliferation under physiological conditions. Of note, this dependency on nucleotide salvage in plasma-like conditions was not caused by precursor metabolite limitation for de novo synthesis. Instead, we found that physiological folate levels are insufficient to support deoxynucleotide triphosphate (dNTP) synthesis for genome replication, leading to DNA replication stress and impaired proliferation when nucleotide salvage is disrupted. Consistently, dietary folate restriction exacerbates the impaired leukemia progression phenotype of nucleotide salvage-deficient B-ALL cells. Together, these findings demonstrate that access to folates is an endogenous limitation for nucleotide synthesis in plasma-like nutrient conditions, increasing the relevance of nucleotide salvage pathways for leukemia progression. More broadly, this work highlights how micronutrient abundance can influence metabolic dependencies and reveals that folate levels shape nucleotide metabolism under physiological conditions.
ER -