Expression profiling in progressive stages of fumarate-hydratase deficiency

The contribution of metabolic changes to tumorigenesis

Houman Ashrafian, Linda O'Flaherty, Julie Adam, Violetta Steeples, Yuen Li Chung, Phil East, Sakari Vanharanta, Heli Lehtonen, Emma Nye, Emine Hatipoglu, Melroy Miranda, Kimberley Howarth, Deepa Shukla, Helen Troy, John Griffiths, Bradley Spencer-Dene, Mohammed Yusuf, Emanuela Volpi, Patrick H. Maxwell, Gordon Stamp & 11 others Richard Poulsom, Christopher W. Pugh, Barbara Costa, Chiara Bardella, Maria Flavia Di Renzo, Michael I. Kotlikoff, Virpi Launonen, Lauri Aaltonen, Mona El-Bahrawy, Ian Tomlinson, Patrick J. Pollard

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is caused by mutations in the Krebs cycle enzyme fumarate hydratase (FH). It has been proposed that "pseudohypoxic" stabilization of hypoxia-inducible factor-α (HIF-α) by fumarate accumulation contributes to tumorigenesis in HLRCC. We hypothesized that an additional direct consequence of FH deficiency is the establishment of a biosynthetic milieu. To investigate this hypothesis, we isolated primary mouse embryonic fibroblast (MEF) lines from Fh1-deficient mice. As predicted, these MEFs upregulated Hif-1α and HIF target genes directly as a result of FH deficiency. In addition, detailed metabolic assessment of these MEFs confirmed their dependence on glycolysis, and an elevated rate of lactate efflux, associated with the upregulation of glycolytic enzymes known to be associated with tumorigenesis. Correspondingly, Fh1-deficient benign murine renal cysts and an advanced human HLRCC-related renal cell carcinoma manifested a prominent and progressive increase in the expression of HIF-á target genes and in genes known to be relevant to tumorigenesis and metastasis. In accord with our hypothesis, in a variety of different FH-deficient tissues, including a novel murine model of Fh1-deficient smooth muscle, we show a striking and progressive upregulation of a tumorigenic metabolic profile, as manifested by increased PKM2 and LDHA protein. Based on the models assessed herein, we infer that that FH deficiency compels cells to adopt an early, reversible, and progressive protumorigenic metabolic milieu that is reminiscent of that driving the Warburg effect. Targets identified in these novel and diverse FH-deficient models represent excellent potential candidates for further mechanistic investigation and therapeutic metabolic manipulation in tumors.

Original languageEnglish
Pages (from-to)9153-9165
Number of pages13
JournalCancer Research
Volume70
Issue number22
DOIs
Publication statusPublished - Nov 15 2010

Fingerprint

Fumarate Hydratase
Leiomyomatosis
Renal Cell Carcinoma
Carcinogenesis
Up-Regulation
Genes
Fumarates
Citric Acid Cycle
Metabolome
Glycolysis
Enzymes
Smooth Muscle
Cysts
Lactic Acid
Fibroblasts
Neoplasm Metastasis
Kidney
Mutation
Fumaric aciduria
Neoplasms

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Ashrafian, H., O'Flaherty, L., Adam, J., Steeples, V., Chung, Y. L., East, P., ... Pollard, P. J. (2010). Expression profiling in progressive stages of fumarate-hydratase deficiency: The contribution of metabolic changes to tumorigenesis. Cancer Research, 70(22), 9153-9165. https://doi.org/10.1158/0008-5472.CAN-10-1949

Expression profiling in progressive stages of fumarate-hydratase deficiency : The contribution of metabolic changes to tumorigenesis. / Ashrafian, Houman; O'Flaherty, Linda; Adam, Julie; Steeples, Violetta; Chung, Yuen Li; East, Phil; Vanharanta, Sakari; Lehtonen, Heli; Nye, Emma; Hatipoglu, Emine; Miranda, Melroy; Howarth, Kimberley; Shukla, Deepa; Troy, Helen; Griffiths, John; Spencer-Dene, Bradley; Yusuf, Mohammed; Volpi, Emanuela; Maxwell, Patrick H.; Stamp, Gordon; Poulsom, Richard; Pugh, Christopher W.; Costa, Barbara; Bardella, Chiara; Di Renzo, Maria Flavia; Kotlikoff, Michael I.; Launonen, Virpi; Aaltonen, Lauri; El-Bahrawy, Mona; Tomlinson, Ian; Pollard, Patrick J.

In: Cancer Research, Vol. 70, No. 22, 15.11.2010, p. 9153-9165.

Research output: Contribution to journalArticle

Ashrafian, H, O'Flaherty, L, Adam, J, Steeples, V, Chung, YL, East, P, Vanharanta, S, Lehtonen, H, Nye, E, Hatipoglu, E, Miranda, M, Howarth, K, Shukla, D, Troy, H, Griffiths, J, Spencer-Dene, B, Yusuf, M, Volpi, E, Maxwell, PH, Stamp, G, Poulsom, R, Pugh, CW, Costa, B, Bardella, C, Di Renzo, MF, Kotlikoff, MI, Launonen, V, Aaltonen, L, El-Bahrawy, M, Tomlinson, I & Pollard, PJ 2010, 'Expression profiling in progressive stages of fumarate-hydratase deficiency: The contribution of metabolic changes to tumorigenesis', Cancer Research, vol. 70, no. 22, pp. 9153-9165. https://doi.org/10.1158/0008-5472.CAN-10-1949
Ashrafian, Houman ; O'Flaherty, Linda ; Adam, Julie ; Steeples, Violetta ; Chung, Yuen Li ; East, Phil ; Vanharanta, Sakari ; Lehtonen, Heli ; Nye, Emma ; Hatipoglu, Emine ; Miranda, Melroy ; Howarth, Kimberley ; Shukla, Deepa ; Troy, Helen ; Griffiths, John ; Spencer-Dene, Bradley ; Yusuf, Mohammed ; Volpi, Emanuela ; Maxwell, Patrick H. ; Stamp, Gordon ; Poulsom, Richard ; Pugh, Christopher W. ; Costa, Barbara ; Bardella, Chiara ; Di Renzo, Maria Flavia ; Kotlikoff, Michael I. ; Launonen, Virpi ; Aaltonen, Lauri ; El-Bahrawy, Mona ; Tomlinson, Ian ; Pollard, Patrick J. / Expression profiling in progressive stages of fumarate-hydratase deficiency : The contribution of metabolic changes to tumorigenesis. In: Cancer Research. 2010 ; Vol. 70, No. 22. pp. 9153-9165.
@article{cd60dd19855541588b951f6473b48b16,
title = "Expression profiling in progressive stages of fumarate-hydratase deficiency: The contribution of metabolic changes to tumorigenesis",
abstract = "Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is caused by mutations in the Krebs cycle enzyme fumarate hydratase (FH). It has been proposed that {"}pseudohypoxic{"} stabilization of hypoxia-inducible factor-α (HIF-α) by fumarate accumulation contributes to tumorigenesis in HLRCC. We hypothesized that an additional direct consequence of FH deficiency is the establishment of a biosynthetic milieu. To investigate this hypothesis, we isolated primary mouse embryonic fibroblast (MEF) lines from Fh1-deficient mice. As predicted, these MEFs upregulated Hif-1α and HIF target genes directly as a result of FH deficiency. In addition, detailed metabolic assessment of these MEFs confirmed their dependence on glycolysis, and an elevated rate of lactate efflux, associated with the upregulation of glycolytic enzymes known to be associated with tumorigenesis. Correspondingly, Fh1-deficient benign murine renal cysts and an advanced human HLRCC-related renal cell carcinoma manifested a prominent and progressive increase in the expression of HIF-{\'a} target genes and in genes known to be relevant to tumorigenesis and metastasis. In accord with our hypothesis, in a variety of different FH-deficient tissues, including a novel murine model of Fh1-deficient smooth muscle, we show a striking and progressive upregulation of a tumorigenic metabolic profile, as manifested by increased PKM2 and LDHA protein. Based on the models assessed herein, we infer that that FH deficiency compels cells to adopt an early, reversible, and progressive protumorigenic metabolic milieu that is reminiscent of that driving the Warburg effect. Targets identified in these novel and diverse FH-deficient models represent excellent potential candidates for further mechanistic investigation and therapeutic metabolic manipulation in tumors.",
author = "Houman Ashrafian and Linda O'Flaherty and Julie Adam and Violetta Steeples and Chung, {Yuen Li} and Phil East and Sakari Vanharanta and Heli Lehtonen and Emma Nye and Emine Hatipoglu and Melroy Miranda and Kimberley Howarth and Deepa Shukla and Helen Troy and John Griffiths and Bradley Spencer-Dene and Mohammed Yusuf and Emanuela Volpi and Maxwell, {Patrick H.} and Gordon Stamp and Richard Poulsom and Pugh, {Christopher W.} and Barbara Costa and Chiara Bardella and {Di Renzo}, {Maria Flavia} and Kotlikoff, {Michael I.} and Virpi Launonen and Lauri Aaltonen and Mona El-Bahrawy and Ian Tomlinson and Pollard, {Patrick J.}",
year = "2010",
month = "11",
day = "15",
doi = "10.1158/0008-5472.CAN-10-1949",
language = "English",
volume = "70",
pages = "9153--9165",
journal = "Journal of Cancer Research",
issn = "0008-5472",
publisher = "American Association for Cancer Research Inc.",
number = "22",

}

TY - JOUR

T1 - Expression profiling in progressive stages of fumarate-hydratase deficiency

T2 - The contribution of metabolic changes to tumorigenesis

AU - Ashrafian, Houman

AU - O'Flaherty, Linda

AU - Adam, Julie

AU - Steeples, Violetta

AU - Chung, Yuen Li

AU - East, Phil

AU - Vanharanta, Sakari

AU - Lehtonen, Heli

AU - Nye, Emma

AU - Hatipoglu, Emine

AU - Miranda, Melroy

AU - Howarth, Kimberley

AU - Shukla, Deepa

AU - Troy, Helen

AU - Griffiths, John

AU - Spencer-Dene, Bradley

AU - Yusuf, Mohammed

AU - Volpi, Emanuela

AU - Maxwell, Patrick H.

AU - Stamp, Gordon

AU - Poulsom, Richard

AU - Pugh, Christopher W.

AU - Costa, Barbara

AU - Bardella, Chiara

AU - Di Renzo, Maria Flavia

AU - Kotlikoff, Michael I.

AU - Launonen, Virpi

AU - Aaltonen, Lauri

AU - El-Bahrawy, Mona

AU - Tomlinson, Ian

AU - Pollard, Patrick J.

PY - 2010/11/15

Y1 - 2010/11/15

N2 - Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is caused by mutations in the Krebs cycle enzyme fumarate hydratase (FH). It has been proposed that "pseudohypoxic" stabilization of hypoxia-inducible factor-α (HIF-α) by fumarate accumulation contributes to tumorigenesis in HLRCC. We hypothesized that an additional direct consequence of FH deficiency is the establishment of a biosynthetic milieu. To investigate this hypothesis, we isolated primary mouse embryonic fibroblast (MEF) lines from Fh1-deficient mice. As predicted, these MEFs upregulated Hif-1α and HIF target genes directly as a result of FH deficiency. In addition, detailed metabolic assessment of these MEFs confirmed their dependence on glycolysis, and an elevated rate of lactate efflux, associated with the upregulation of glycolytic enzymes known to be associated with tumorigenesis. Correspondingly, Fh1-deficient benign murine renal cysts and an advanced human HLRCC-related renal cell carcinoma manifested a prominent and progressive increase in the expression of HIF-á target genes and in genes known to be relevant to tumorigenesis and metastasis. In accord with our hypothesis, in a variety of different FH-deficient tissues, including a novel murine model of Fh1-deficient smooth muscle, we show a striking and progressive upregulation of a tumorigenic metabolic profile, as manifested by increased PKM2 and LDHA protein. Based on the models assessed herein, we infer that that FH deficiency compels cells to adopt an early, reversible, and progressive protumorigenic metabolic milieu that is reminiscent of that driving the Warburg effect. Targets identified in these novel and diverse FH-deficient models represent excellent potential candidates for further mechanistic investigation and therapeutic metabolic manipulation in tumors.

AB - Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is caused by mutations in the Krebs cycle enzyme fumarate hydratase (FH). It has been proposed that "pseudohypoxic" stabilization of hypoxia-inducible factor-α (HIF-α) by fumarate accumulation contributes to tumorigenesis in HLRCC. We hypothesized that an additional direct consequence of FH deficiency is the establishment of a biosynthetic milieu. To investigate this hypothesis, we isolated primary mouse embryonic fibroblast (MEF) lines from Fh1-deficient mice. As predicted, these MEFs upregulated Hif-1α and HIF target genes directly as a result of FH deficiency. In addition, detailed metabolic assessment of these MEFs confirmed their dependence on glycolysis, and an elevated rate of lactate efflux, associated with the upregulation of glycolytic enzymes known to be associated with tumorigenesis. Correspondingly, Fh1-deficient benign murine renal cysts and an advanced human HLRCC-related renal cell carcinoma manifested a prominent and progressive increase in the expression of HIF-á target genes and in genes known to be relevant to tumorigenesis and metastasis. In accord with our hypothesis, in a variety of different FH-deficient tissues, including a novel murine model of Fh1-deficient smooth muscle, we show a striking and progressive upregulation of a tumorigenic metabolic profile, as manifested by increased PKM2 and LDHA protein. Based on the models assessed herein, we infer that that FH deficiency compels cells to adopt an early, reversible, and progressive protumorigenic metabolic milieu that is reminiscent of that driving the Warburg effect. Targets identified in these novel and diverse FH-deficient models represent excellent potential candidates for further mechanistic investigation and therapeutic metabolic manipulation in tumors.

UR - http://www.scopus.com/inward/record.url?scp=78549275858&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78549275858&partnerID=8YFLogxK

U2 - 10.1158/0008-5472.CAN-10-1949

DO - 10.1158/0008-5472.CAN-10-1949

M3 - Article

VL - 70

SP - 9153

EP - 9165

JO - Journal of Cancer Research

JF - Journal of Cancer Research

SN - 0008-5472

IS - 22

ER -