Proline synthesis through PYCR1 is required to support cancer cell proliferation and survival in oxygen-limiting conditions

Rebecca Westbrook; Esther Bridges; Jennie Roberts; Cristina Escribano Gonzalez; Katherine Eales; Lisa Vettore; Paul Walker; Elias Vera-Siguenza; Himani Rana; Federica Cuozzo; Kattri-Liis Eskla; Hans Vellama; Abeer Shaaban; Colin Nixon; Hendrik Luuk; Gareth Lavery; David Hodson; Adrian L Harris; Daniel Tennant

doi:10.1016/j.celrep.2022.110320

Proline synthesis through PYCR1 is required to support cancer cell proliferation and survival in oxygen-limiting conditions

Rebecca Westbrook, Esther Bridges, Jennie Roberts, Cristina Escribano Gonzalez, Katherine Eales, Lisa Vettore, Paul Walker, Elias Vera-Siguenza, Himani Rana, Federica Cuozzo, Kattri-Liis Eskla, Hans Vellama, Abeer Shaaban, Colin Nixon, Hendrik Luuk, Gareth Lavery, David Hodson, Adrian L Harris, Daniel Tennant

Metabolism and Systems Research

Research output: Contribution to journal › Article › peer-review

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Abstract

The demands of cancer cell proliferation alongside an inadequate angiogenic response lead to insufficient oxygen availability in the tumor microenvironment. Within the mitochondria, oxygen is the major electron acceptor for NADH, with the result that the reducing potential produced through tricarboxylic acid (TCA) cycle activity and mitochondrial respiration are functionally linked. As the oxidizing activity of the TCA cycle is required for efficient synthesis of anabolic precursors, tumoral hypoxia could lead to a cessation of proliferation without another means of correcting the redox imbalance. We show that in hypoxic conditions, mitochondrial pyrroline 5-carboxylate reductase 1 (PYCR1) activity is increased, oxidizing NADH with the synthesis of proline as a by-product. We further show that PYCR1 activity is required for the successful maintenance of hypoxic regions by permitting continued TCA cycle activity, and that its loss leads to significantly increased hypoxia in vivo and in 3D culture, resulting in widespread cell death.

Original language	English
Article number	110320
Number of pages	15
Journal	Cell Reports
Volume	38
Issue number	5
DOIs	https://doi.org/10.1016/j.celrep.2022.110320
Publication status	Published - 1 Feb 2022

Bibliographical note

Funding Information:
The authors would like to acknowledge funding from Cancer Research UK to R.L.W. C.E.G. K.L.E. E.V.-S. L.A.V. and P.D.W. (C42109/A26982 and C42109/A24747). We would also like to acknowledge Professor Reuven Agami, who kindly gifted the PYCR1?/? line and control. The graphical abstract was created with BioRender.com. Finally, we would like to acknowledge the support and resources of the Birmingham Metabolic Tracer Analysis Core. R.L.W. E.B. A.S. H.L. A.L.H. and D.A.T. conceptualized the study and designed the methodology. R.L.W. E.B. C.E.G. J.R. H.R. K.L.E. K.-L.E. H.V. E.V.-S. L.A.V. P.D.W. F.C. and C.N. performed investigations. E.V.-S. E.B. R.L.W. K.-L.E. H.V. C.N. and D.A.T. performed formal analysis and data visualization. G.G.L. D.J.H. A.S. H.L. C.N. A.L.H. and D.A.T. provided resources. R.L.W. E.B. J.B. C.N. and D.A.T. wrote the original manuscript draft, while all authors contributed to reviewing and editing of the manuscript. The authors declare no competing interests.

Funding Information:
The authors would like to acknowledge funding from Cancer Research UK to R.L.W., C.E.G., K.L.E., E.V.-S., L.A.V., and P.D.W. (C42109/ A26982 and C42109/ A24747 ). We would also like to acknowledge Professor Reuven Agami, who kindly gifted the PYCR1 −/− line and control. The graphical abstract was created with BioRender.com. Finally, we would like to acknowledge the support and resources of the Birmingham Metabolic Tracer Analysis Core.

Publisher Copyright:
© 2022 The Authors

Keywords

hypoxia
proline
PYCR1
redox
cancer
mitochondria
NADH

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.celrep.2022.110320Licence: Creative Commons: Attribution (CC BY)

WestbrookR2022ProlineFinal published version, 5.11 MBLicence: Creative Commons: Attribution (CC BY)

https://www.cell.com/cell-reports/fulltext/S2211-1247(22)00031-6Licence: Creative Commons: Attribution (CC BY)

Pseudohypoxia-led identification of metabolic vulnerabilities in hypoxic tumours to improve patient outcomes
Tennant, D. & Ludwig, C.
Cancer Research Uk
1/10/18 → 30/09/22
Project: Research
Pseudohypoxia-led identification of metabolic vulnerabilities in hypoxic tumours to improve patient outcomes
Tennant, D., Ludwig, C. & Spill, F.
Cancer Research Uk
1/02/18 → 31/01/24
Project: Research

Cite this

Westbrook, R., Bridges, E., Roberts, J., Escribano Gonzalez, C., Eales, K., Vettore, L., Walker, P., Vera-Siguenza, E., Rana, H., Cuozzo, F., Eskla, K.-L., Vellama, H., Shaaban, A., Nixon, C., Luuk, H., Lavery, G., Hodson, D., Harris, A. L., & Tennant, D. (2022). Proline synthesis through PYCR1 is required to support cancer cell proliferation and survival in oxygen-limiting conditions. Cell Reports, 38(5), Article 110320. https://doi.org/10.1016/j.celrep.2022.110320

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title = "Proline synthesis through PYCR1 is required to support cancer cell proliferation and survival in oxygen-limiting conditions",

abstract = "The demands of cancer cell proliferation alongside an inadequate angiogenic response lead to insufficient oxygen availability in the tumor microenvironment. Within the mitochondria, oxygen is the major electron acceptor for NADH, with the result that the reducing potential produced through tricarboxylic acid (TCA) cycle activity and mitochondrial respiration are functionally linked. As the oxidizing activity of the TCA cycle is required for efficient synthesis of anabolic precursors, tumoral hypoxia could lead to a cessation of proliferation without another means of correcting the redox imbalance. We show that in hypoxic conditions, mitochondrial pyrroline 5-carboxylate reductase 1 (PYCR1) activity is increased, oxidizing NADH with the synthesis of proline as a by-product. We further show that PYCR1 activity is required for the successful maintenance of hypoxic regions by permitting continued TCA cycle activity, and that its loss leads to significantly increased hypoxia in vivo and in 3D culture, resulting in widespread cell death.",

keywords = "hypoxia, proline, PYCR1, redox, cancer, mitochondria, NADH",

author = "Rebecca Westbrook and Esther Bridges and Jennie Roberts and {Escribano Gonzalez}, Cristina and Katherine Eales and Lisa Vettore and Paul Walker and Elias Vera-Siguenza and Himani Rana and Federica Cuozzo and Kattri-Liis Eskla and Hans Vellama and Abeer Shaaban and Colin Nixon and Hendrik Luuk and Gareth Lavery and David Hodson and Harris, {Adrian L} and Daniel Tennant",

note = "Funding Information: The authors would like to acknowledge funding from Cancer Research UK to R.L.W. C.E.G. K.L.E. E.V.-S. L.A.V. and P.D.W. (C42109/A26982 and C42109/A24747). We would also like to acknowledge Professor Reuven Agami, who kindly gifted the PYCR1?/? line and control. The graphical abstract was created with BioRender.com. Finally, we would like to acknowledge the support and resources of the Birmingham Metabolic Tracer Analysis Core. R.L.W. E.B. A.S. H.L. A.L.H. and D.A.T. conceptualized the study and designed the methodology. R.L.W. E.B. C.E.G. J.R. H.R. K.L.E. K.-L.E. H.V. E.V.-S. L.A.V. P.D.W. F.C. and C.N. performed investigations. E.V.-S. E.B. R.L.W. K.-L.E. H.V. C.N. and D.A.T. performed formal analysis and data visualization. G.G.L. D.J.H. A.S. H.L. C.N. A.L.H. and D.A.T. provided resources. R.L.W. E.B. J.B. C.N. and D.A.T. wrote the original manuscript draft, while all authors contributed to reviewing and editing of the manuscript. The authors declare no competing interests. Funding Information: The authors would like to acknowledge funding from Cancer Research UK to R.L.W., C.E.G., K.L.E., E.V.-S., L.A.V., and P.D.W. (C42109/ A26982 and C42109/ A24747 ). We would also like to acknowledge Professor Reuven Agami, who kindly gifted the PYCR1 −/− line and control. The graphical abstract was created with BioRender.com. Finally, we would like to acknowledge the support and resources of the Birmingham Metabolic Tracer Analysis Core. Publisher Copyright: {\textcopyright} 2022 The Authors",

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Westbrook, R, Bridges, E, Roberts, J, Escribano Gonzalez, C , Eales, K , Vettore, L, Walker, P, Vera-Siguenza, E, Rana, H, Cuozzo, F, Eskla, K-L, Vellama, H, Shaaban, A, Nixon, C, Luuk, H, Lavery, G, Hodson, D, Harris, AL & Tennant, D 2022, 'Proline synthesis through PYCR1 is required to support cancer cell proliferation and survival in oxygen-limiting conditions', Cell Reports, vol. 38, no. 5, 110320. https://doi.org/10.1016/j.celrep.2022.110320

TY - JOUR

T1 - Proline synthesis through PYCR1 is required to support cancer cell proliferation and survival in oxygen-limiting conditions

AU - Westbrook, Rebecca

AU - Bridges, Esther

AU - Roberts, Jennie

AU - Escribano Gonzalez, Cristina

AU - Eales, Katherine

AU - Vettore, Lisa

AU - Walker, Paul

AU - Vera-Siguenza, Elias

AU - Rana, Himani

AU - Cuozzo, Federica

AU - Eskla, Kattri-Liis

AU - Vellama, Hans

AU - Shaaban, Abeer

AU - Nixon, Colin

AU - Luuk, Hendrik

AU - Lavery, Gareth

AU - Hodson, David

AU - Harris, Adrian L

AU - Tennant, Daniel

N1 - Funding Information: The authors would like to acknowledge funding from Cancer Research UK to R.L.W. C.E.G. K.L.E. E.V.-S. L.A.V. and P.D.W. (C42109/A26982 and C42109/A24747). We would also like to acknowledge Professor Reuven Agami, who kindly gifted the PYCR1?/? line and control. The graphical abstract was created with BioRender.com. Finally, we would like to acknowledge the support and resources of the Birmingham Metabolic Tracer Analysis Core. R.L.W. E.B. A.S. H.L. A.L.H. and D.A.T. conceptualized the study and designed the methodology. R.L.W. E.B. C.E.G. J.R. H.R. K.L.E. K.-L.E. H.V. E.V.-S. L.A.V. P.D.W. F.C. and C.N. performed investigations. E.V.-S. E.B. R.L.W. K.-L.E. H.V. C.N. and D.A.T. performed formal analysis and data visualization. G.G.L. D.J.H. A.S. H.L. C.N. A.L.H. and D.A.T. provided resources. R.L.W. E.B. J.B. C.N. and D.A.T. wrote the original manuscript draft, while all authors contributed to reviewing and editing of the manuscript. The authors declare no competing interests. Funding Information: The authors would like to acknowledge funding from Cancer Research UK to R.L.W., C.E.G., K.L.E., E.V.-S., L.A.V., and P.D.W. (C42109/ A26982 and C42109/ A24747 ). We would also like to acknowledge Professor Reuven Agami, who kindly gifted the PYCR1 −/− line and control. The graphical abstract was created with BioRender.com. Finally, we would like to acknowledge the support and resources of the Birmingham Metabolic Tracer Analysis Core. Publisher Copyright: © 2022 The Authors

PY - 2022/2/1

Y1 - 2022/2/1

N2 - The demands of cancer cell proliferation alongside an inadequate angiogenic response lead to insufficient oxygen availability in the tumor microenvironment. Within the mitochondria, oxygen is the major electron acceptor for NADH, with the result that the reducing potential produced through tricarboxylic acid (TCA) cycle activity and mitochondrial respiration are functionally linked. As the oxidizing activity of the TCA cycle is required for efficient synthesis of anabolic precursors, tumoral hypoxia could lead to a cessation of proliferation without another means of correcting the redox imbalance. We show that in hypoxic conditions, mitochondrial pyrroline 5-carboxylate reductase 1 (PYCR1) activity is increased, oxidizing NADH with the synthesis of proline as a by-product. We further show that PYCR1 activity is required for the successful maintenance of hypoxic regions by permitting continued TCA cycle activity, and that its loss leads to significantly increased hypoxia in vivo and in 3D culture, resulting in widespread cell death.

AB - The demands of cancer cell proliferation alongside an inadequate angiogenic response lead to insufficient oxygen availability in the tumor microenvironment. Within the mitochondria, oxygen is the major electron acceptor for NADH, with the result that the reducing potential produced through tricarboxylic acid (TCA) cycle activity and mitochondrial respiration are functionally linked. As the oxidizing activity of the TCA cycle is required for efficient synthesis of anabolic precursors, tumoral hypoxia could lead to a cessation of proliferation without another means of correcting the redox imbalance. We show that in hypoxic conditions, mitochondrial pyrroline 5-carboxylate reductase 1 (PYCR1) activity is increased, oxidizing NADH with the synthesis of proline as a by-product. We further show that PYCR1 activity is required for the successful maintenance of hypoxic regions by permitting continued TCA cycle activity, and that its loss leads to significantly increased hypoxia in vivo and in 3D culture, resulting in widespread cell death.

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KW - proline

KW - PYCR1

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KW - cancer

KW - mitochondria

KW - NADH

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DO - 10.1016/j.celrep.2022.110320

M3 - Article

C2 - 35108535

SN - 2211-1247

VL - 38

JO - Cell Reports

JF - Cell Reports

IS - 5

M1 - 110320

ER -

Proline synthesis through PYCR1 is required to support cancer cell proliferation and survival in oxygen-limiting conditions

Abstract

Bibliographical note

Keywords

UN SDGs

Access to Document

Fingerprint

Projects

Pseudohypoxia-led identification of metabolic vulnerabilities in hypoxic tumours to improve patient outcomes

Pseudohypoxia-led identification of metabolic vulnerabilities in hypoxic tumours to improve patient outcomes

Cite this