Cell Culture Medium Formulation and Its Implications in Cancer Metabolism

Ackermann, Tobias; Tardito, Saverio

doi:10.1016/j.trecan.2019.05.004

Cited by 113 publications

(91 citation statements)

References 16 publications

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“…Interestingly, we also found that TLR3 but not TLR4 associated mitochondrial reprogramming was dependent on glucose availability in the microenvironment. Supra-physiological levels of glucose have been shown to decrease a cell's dependence on mitochondria for energy production 52,64 . Similarly, our results suggest high glucose conditions may also alter mitochondr ia l reprogramming associated with TLR3 engagement.…”

mentioning

confidence: 99%

Differential remodeling of the electron transport chain is required to support TLR3 and TLR4 signaling and cytokine production in macrophages

Ahmed

Roy

Jaworski

et al. 2019

Preprint

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Increasing evidence suggests that mitochondria play a critical role in driving innate immune responses against bacteria and viruses. However, it is unclear if differential reprogramming of mitochondrial function contributes to the fine tuning of pathogen specific immune responses. Here, we found that TLR3 and TLR4 engagement on murine bone marrow derived macrophages was associated with differential remodeling of electron transport chain complex expression. This remodeling was associated with differential accumulation of mitochondrial and cytosolic ROS, which were required to support ligand specific inflammatory and antiviral cytokine production.We also found that the magnitude of TLR3, but not TLR4, responses were modulated by glucose availability. Under conditions of low glucose conditions, TLR3 engagement was associated with increased ETC complex III expression, increased mitochondrial and cytosolic ROS and increased inflammatory and antiviral cytokine production. This amplification was selectively reversed by targeting superoxide production from the outer Q-binding site of the ETC complex III. These results suggest that ligand specific modulation of the ETC may act as a rheostat that fine-tunes innate immune responses via mitochondrial ROS production. Modulation of these processes may represent a novel mechanism to modulate the nature as well as the magnitude of antiviral versus inflammatory immune responses.3 Introduction:

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mentioning

confidence: 99%

Differential remodeling of the electron transport chain is required to support TLR3 and TLR4 signaling and cytokine production in macrophages

Ahmed

Roy

Jaworski

et al. 2019

Preprint

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“…A combination of the above explanations most likely contributes to the cultured cells versus primary tumor discrepancy. It is already well-established that cell culture conditions do not always accurately reflect physiological conditions (Ackermann and Tardito, 2019;Sanderson et al, 2019;Vande Voorde et al, 2019). Prior to in vitro metabolomics profiling, we cultured our cells using heat-inactivated fetal bovine serum which, thereby nullifying any MTAP-activity that would otherwise be present in the serum.…”

Section: Discussionmentioning

confidence: 99%

Methylthioadenosine is Not Dramatically Elevated inMTAP-Homozygous Deleted Primary Glioblastomas

Barekatain

Yan

Ackroyd

et al. 2019

Preprint

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Highlights• Methylthioadenosine (MTA) is elevated in MTAP-deleted cancer cells in vitro, which provides a selective vulnerability to PRMT5 and MAT2A inhibitors • Accumulation of MTA in MTAP-deleted cancer cells is predominately extracellular, suggesting active secretion of MTA.• MTAP-deleted primary human glioblastoma tumors show minimal intratumoral elevations of MTA, which is likely explained by secretion and metabolism by MTAP-competent stromal cells. SUMMARYHomozygous deletion of the CDK2NA locus frequently results in co-deletion of methylthioadenosine phosphorylase (MTAP) in many fatal cancers such as glioblastoma multiforme (GBM), resulting in elevations of the substrate metabolite, methylthioadenosine (MTA). To capitalize on such accumulations, therapeutic targeting of protein arginine methyltransferase 5 (PRMT5) and methionine adenosyl transferase (MAT2A) are ongoing. While extensively corroborated in vitro, the clinical efficacy of these strategies ultimately relies on equally significant accumulations of MTA in human tumors.Here, we show that in vitro accumulation of MTA is a predominately extracellular phenomenon, indicating secretion of MTA from MTAP-deleted cells. In primary human GBMs, we find that MTA levels are not significantly higher in MTAP-deleted compared to MTAP-intact tumors or normal brain tissue. Together, these findings highlight the metabolic discrepancies between in vitro models and primary human tumors and should thus be carefully considered in the development of the precision therapies targeting MTAP-homozygous deleted GBM.

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“…Moreover, in vitro screens of human cells have been performed in traditional media that poorly recapitulate metabolite availability in human blood. (Ackermann and Tardito, 2019;Cantor, 2019).…”

Section: Introductionmentioning

confidence: 99%

CRISPR screens in physiologic medium reveal conditionally essential genes in human cells

Rossiter

Huggler

Adelmann

et al. 2020

Preprint

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SUMMARYForward genetic screens across hundreds of diverse cancer cell lines have started to define the genetic dependencies of proliferating human cells and how these vary by genotype and lineage. Most screens, however, have been carried out in culture media that poorly resemble metabolite availability in human blood. To explore how medium composition influences gene essentiality, we performed CRISPR-based screens of human cancer cell lines cultured in traditional versus human plasma-like medium (HPLM). Sets of medium-dependent fitness genes span several cellular processes and can vary with both natural cell-intrinsic diversity and the specific combination of basal and serum components that comprise typical culture media. Notably, we traced the causes for each of three conditional growth phenotypes to the availability of metabolites uniquely defined in HPLM versus traditional media. Our findings reveal the profound impact of medium composition on gene essentiality in human cells, and also suggest general strategies for using genetic screens in HPLM to uncover new cancer vulnerabilities and gene-nutrient interactions.

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Cell Culture Medium Formulation and Its Implications in Cancer Metabolism

Cited by 113 publications

References 16 publications

Differential remodeling of the electron transport chain is required to support TLR3 and TLR4 signaling and cytokine production in macrophages

Differential remodeling of the electron transport chain is required to support TLR3 and TLR4 signaling and cytokine production in macrophages

Methylthioadenosine is Not Dramatically Elevated inMTAP-Homozygous Deleted Primary Glioblastomas

CRISPR screens in physiologic medium reveal conditionally essential genes in human cells

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