Untargeted Lipidomics Reveals Dynamic Changes to the HEK 293T Lipidome During Lentiviral Vector Production Over 72 Hours
Abstract
Lentiviral vectors (LV) are biotechnology tools descending from human immunodeficiency virus (HIV) that may be used in cell and gene therapies. HIV transduction efficiency has been shown to be negatively affected when the lipid composition of its envelope is altered, suggesting that targeted lipid envelope composition engineering may serve as a tool to modulate transduction. To investigate this possibility, an untargeted lipidomics experiment was conducted on HEK 293T packaging cells producing LV at three time points: 24, 48 and 72 h. 401 lipids from 23 classes were identified and significant changes to the HEK 293T lipidome from LV production were recorded at each time point. Triacylglycerol (TG) and phosphatidylcholine (PC) species were the most affected lipid classes throughout the time course. Large increases in the abundance of TG species were observed with significant depletion of many PC species, suggesting lipid droplet formation occurs during LV production. Analysis of the fatty acyl chain profiles revealed that LV production significantly increased the average chain length after 24 and 48 h while the double bond index did not change in a statistically significant manner across all time points compared to control cells, possibly serving as a means to maintain homeostasis while large amounts of shorter acyl chains were used for LV envelope formation. 9 of 12 acyl chains had significant trends with increased LV production time, serving as targets for media supplementation or for metabolic pathway manipulation to increase LV titer. Future directions will investigate the effects of media supplementation with phospholipid mixtures on the dynamics of TG species over time with LV production.
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BibTeX
@misc{roberts2026untargeted,
title = {{Untargeted Lipidomics Reveals Dynamic Changes to the HEK 293T Lipidome During Lentiviral Vector Production Over 72 Hours}},
author = {Joshua. A. Roberts and Elena Godbout and Jocelyn. A. Menard and Christopher. N. Boddy and Jean-Simon Diallo and Jeffrey. C. Smith},
howpublished = {ChemRxiv},
year = {2026},
month = oct,
doi = {10.26434/chemrxiv.15011015/v1},
url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15011015/v1},
note = {CC BY-NC},
}
RIS
TY - UNPB
TI - Untargeted Lipidomics Reveals Dynamic Changes to the HEK 293T Lipidome During Lentiviral Vector Production Over 72 Hours
AU - Joshua. A. Roberts
AU - Elena Godbout
AU - Jocelyn. A. Menard
AU - Christopher. N. Boddy
AU - Jean-Simon Diallo
AU - Jeffrey. C. Smith
PY - 2026
DA - 2026/10/25
PB - ChemRxiv
DO - 10.26434/chemrxiv.15011015/v1
UR - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15011015/v1
AB - Lentiviral vectors (LV) are biotechnology tools descending from human immunodeficiency virus (HIV) that may be used in cell and gene therapies. HIV transduction efficiency has been shown to be negatively affected when the lipid composition of its envelope is altered, suggesting that targeted lipid envelope composition engineering may serve as a tool to modulate transduction. To investigate this possibility, an untargeted lipidomics experiment was conducted on HEK 293T packaging cells producing LV at three time points: 24, 48 and 72 h. 401 lipids from 23 classes were identified and significant changes to the HEK 293T lipidome from LV production were recorded at each time point. Triacylglycerol (TG) and phosphatidylcholine (PC) species were the most affected lipid classes throughout the time course. Large increases in the abundance of TG species were observed with significant depletion of many PC species, suggesting lipid droplet formation occurs during LV production. Analysis of the fatty acyl chain profiles revealed that LV production significantly increased the average chain length after 24 and 48 h while the double bond index did not change in a statistically significant manner across all time points compared to control cells, possibly serving as a means to maintain homeostasis while large amounts of shorter acyl chains were used for LV envelope formation. 9 of 12 acyl chains had significant trends with increased LV production time, serving as targets for media supplementation or for metabolic pathway manipulation to increase LV titer. Future directions will investigate the effects of media supplementation with phospholipid mixtures on the dynamics of TG species over time with LV production.
ER -