Detection of glucose-derived d- and l-lactate in cancer cells by the use of a chiral NMR shift reagent

Abstract Background Excessive lactate production, a hallmark of cancer, is largely formed by the reduction of pyruvate via lactate dehydrogenase (LDH) to l-lactate. Although d-lactate can also be produced from glucose via the methylglyoxal pathway in small amounts, less is known about the amount of...

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Autores principales:	Eul Hyun Suh, Carlos F. G. C. Geraldes, Sara Chirayil, Brandon Faubert, Raul Ayala, Ralph J. DeBerardinis, A. Dean Sherry
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	d- and l-lactate Shift reagent-aided NMR Cancer cells Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282
Acceso en línea:	https://doaj.org/article/aa4e4f7bf1184b3ea910f6a8758f378c
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spelling	oai:doaj.org-article:aa4e4f7bf1184b3ea910f6a8758f378c2021-11-08T11:13:00ZDetection of glucose-derived d- and l-lactate in cancer cells by the use of a chiral NMR shift reagent10.1186/s40170-021-00267-42049-3002https://doaj.org/article/aa4e4f7bf1184b3ea910f6a8758f378c2021-11-01T00:00:00Zhttps://doi.org/10.1186/s40170-021-00267-4https://doaj.org/toc/2049-3002Abstract Background Excessive lactate production, a hallmark of cancer, is largely formed by the reduction of pyruvate via lactate dehydrogenase (LDH) to l-lactate. Although d-lactate can also be produced from glucose via the methylglyoxal pathway in small amounts, less is known about the amount of d-lactate produced in cancer cells. Since the stereoisomers of lactate cannot be distinguished by conventional 1H NMR spectroscopy, a chiral NMR shift reagent was used to fully resolve the 1H NMR resonances of d- and l-lactate. Methods The production of l-lactate from glucose and d-lactate from methylglyoxal was first demonstrated in freshly isolated red blood cells using the chiral NMR shift reagent, YbDO3A-trisamide. Then, two different cell lines with high GLO1 expression (H1648 and H 1395) were selected from a panel of over 80 well-characterized human NSCLC cell lines, grown to confluence in standard tissue culture media, washed with phosphate-buffered saline, and exposed to glucose in a buffer for 4 h. After 4 h, a small volume of extracellular fluid was collected and mixed with YbDO3A-trisamide for analysis by 1H NMR spectroscopy. Results A suspension of freshly isolated red blood cells exposed to 5mM glucose produced l-lactate as expected but very little d-lactate. To evaluate the utility of the chiral NMR shift reagent, methylglyoxal was then added to red cells along with glucose to stimulate the production of d-lactate via the glyoxalate pathway. In this case, both d-lactate and l-lactate were produced and their NMR chemical shifts assigned. NSCLC cell lines with different expression levels of GLO1 produced both l- and d-lactate after incubation with glucose and glutamine alone. A GLO1-deleted parental cell line (3553T3) showed no production of d-lactate from glucose while re-expression of GLO1 resulted in higher production of d-lactate. Conclusions The shift-reagent-aided NMR technique demonstrates that d-lactate is produced from glucose in NSCLC cells via the methylglyoxal pathway. The biological role of d-lactate is uncertain but a convenient method for monitoring d-lactate production could provide new insights into the biological roles of d- versus l-lactate in cancer metabolism.Eul Hyun SuhCarlos F. G. C. GeraldesSara ChirayilBrandon FaubertRaul AyalaRalph J. DeBerardinisA. Dean SherryBMCarticled- and l-lactateShift reagent-aided NMRCancer cellsNeoplasms. Tumors. Oncology. Including cancer and carcinogensRC254-282ENCancer & Metabolism, Vol 9, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	d- and l-lactate Shift reagent-aided NMR Cancer cells Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282
spellingShingle	d- and l-lactate Shift reagent-aided NMR Cancer cells Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282 Eul Hyun Suh Carlos F. G. C. Geraldes Sara Chirayil Brandon Faubert Raul Ayala Ralph J. DeBerardinis A. Dean Sherry Detection of glucose-derived d- and l-lactate in cancer cells by the use of a chiral NMR shift reagent
description	Abstract Background Excessive lactate production, a hallmark of cancer, is largely formed by the reduction of pyruvate via lactate dehydrogenase (LDH) to l-lactate. Although d-lactate can also be produced from glucose via the methylglyoxal pathway in small amounts, less is known about the amount of d-lactate produced in cancer cells. Since the stereoisomers of lactate cannot be distinguished by conventional 1H NMR spectroscopy, a chiral NMR shift reagent was used to fully resolve the 1H NMR resonances of d- and l-lactate. Methods The production of l-lactate from glucose and d-lactate from methylglyoxal was first demonstrated in freshly isolated red blood cells using the chiral NMR shift reagent, YbDO3A-trisamide. Then, two different cell lines with high GLO1 expression (H1648 and H 1395) were selected from a panel of over 80 well-characterized human NSCLC cell lines, grown to confluence in standard tissue culture media, washed with phosphate-buffered saline, and exposed to glucose in a buffer for 4 h. After 4 h, a small volume of extracellular fluid was collected and mixed with YbDO3A-trisamide for analysis by 1H NMR spectroscopy. Results A suspension of freshly isolated red blood cells exposed to 5mM glucose produced l-lactate as expected but very little d-lactate. To evaluate the utility of the chiral NMR shift reagent, methylglyoxal was then added to red cells along with glucose to stimulate the production of d-lactate via the glyoxalate pathway. In this case, both d-lactate and l-lactate were produced and their NMR chemical shifts assigned. NSCLC cell lines with different expression levels of GLO1 produced both l- and d-lactate after incubation with glucose and glutamine alone. A GLO1-deleted parental cell line (3553T3) showed no production of d-lactate from glucose while re-expression of GLO1 resulted in higher production of d-lactate. Conclusions The shift-reagent-aided NMR technique demonstrates that d-lactate is produced from glucose in NSCLC cells via the methylglyoxal pathway. The biological role of d-lactate is uncertain but a convenient method for monitoring d-lactate production could provide new insights into the biological roles of d- versus l-lactate in cancer metabolism.
format	article
author	Eul Hyun Suh Carlos F. G. C. Geraldes Sara Chirayil Brandon Faubert Raul Ayala Ralph J. DeBerardinis A. Dean Sherry
author_facet	Eul Hyun Suh Carlos F. G. C. Geraldes Sara Chirayil Brandon Faubert Raul Ayala Ralph J. DeBerardinis A. Dean Sherry
author_sort	Eul Hyun Suh
title	Detection of glucose-derived d- and l-lactate in cancer cells by the use of a chiral NMR shift reagent
title_short	Detection of glucose-derived d- and l-lactate in cancer cells by the use of a chiral NMR shift reagent
title_full	Detection of glucose-derived d- and l-lactate in cancer cells by the use of a chiral NMR shift reagent
title_fullStr	Detection of glucose-derived d- and l-lactate in cancer cells by the use of a chiral NMR shift reagent
title_full_unstemmed	Detection of glucose-derived d- and l-lactate in cancer cells by the use of a chiral NMR shift reagent
title_sort	detection of glucose-derived d- and l-lactate in cancer cells by the use of a chiral nmr shift reagent
publisher	BMC
publishDate	2021
url	https://doaj.org/article/aa4e4f7bf1184b3ea910f6a8758f378c
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Detection of glucose-derived d- and l-lactate in cancer cells by the use of a chiral NMR shift reagent

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