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Cellular and Molecular Biology

Dysregulation at multiple points of the kynurenine pathway is a ubiquitous feature of renal cancer: implications for tumour immune evasion

Abstract

Background

Indoleamine 2,3-dioxygenase (IDO), the first step in the kynurenine pathway (KP), is upregulated in some cancers and represents an attractive therapeutic target given its role in tumour immune evasion. However, the recent failure of an IDO inhibitor in a late phase trial raises questions about this strategy.

Methods

Matched renal cell carcinoma (RCC) and normal kidney tissues were subject to proteomic profiling. Tissue immunohistochemistry and gene expression data were used to validate findings. Phenotypic effects of loss/gain of expression were examined in vitro.

Results

Quinolate phosphoribosyltransferase (QPRT), the final and rate-limiting enzyme in the KP, was identified as being downregulated in RCC. Loss of QPRT expression led to increased potential for anchorage-independent growth. Gene expression, mass spectrometry (clear cell and chromophobe RCC) and tissue immunohistochemistry (clear cell, papillary and chromophobe), confirmed loss or decreased expression of QPRT and showed downregulation of other KP enzymes, including kynurenine 3-monoxygenase (KMO) and 3-hydroxyanthranilate-3,4-dioxygenase (HAAO), with a concomitant maintenance or upregulation of nicotinamide phosphoribosyltransferase (NAMPT), the key enzyme in the NAD+ salvage pathway.

Conclusions

Widespread dysregulation of the KP is common in RCC and is likely to contribute to tumour immune evasion, carrying implications for effective therapeutic targeting of this critical pathway

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Fig. 1: QPRT expression is lost in clear cell RCC (ccRCC).
Fig. 2: Loss of QPRT expression increases anchorage-independent growth.
Fig. 3: Multiple proteins in the kynurenine pathway are dysregulated in RCC.
Fig. 4: Gene expression of QPRT, KMO, HAAO and NAMPT is similarly dysregulated in clear cell RCC.
Fig. 5: Alterations to the kynurenine pathway in RCC.
Fig. 6: Immunohistochemical analysis of kynurenine pathway enzymes.

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Acknowledgements

We are grateful to the patients for donating samples and the staff of the Leeds Biobanking and Sample Processing Lab and Leeds Multidisciplinary RTB for samples and Anke Bruning-Richardson for technical support with the TMA. We thank the staff of the oncology and urology departments of Leeds Teaching Hospitals NHS Trust who assisted in patient recruitment, Professor W Kaelin for the 786-VHL cell lines, Robert Schwarz for providing antibody to QPRT, Darren Tomlinson for providing the parental QPRT vector, Anthea Stanley for laboratory support and Nick Totty and Sarah Hanrahan for the sequencing of the 2D-PAGE proteins.

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Contributions

R.E.B., P.S.J. and N.S.V. supervised the study with A.H. and E.R.M. also contributing to study design; R.A.C. and R.E.B. conducted the initial 2D-PAGE work; J. Brown facilitated access to tissue specimens and clinical data; K.R.D., S.T., R.S. and A.Z. conducted the mass spectrometric studies; K.R.D., M.K. and J. Burns conducted and oversaw phenotypic studies; T.F. performed the assay for tissue QUIN concentrations; M.S. and H.S. performed the IHC; S.B. and M.M. provided expert pathological review; L.F., A.B., Y.R., M.L. and G.S. provided transcriptomic data sets; N.H. analysed the data and, together with R.A.C., N.S.V. and R.E.B., drafted the manuscript; all authors provided comments on the manuscript and had final approval of the submitted version.

Corresponding author

Correspondence to Naveen S. Vasudev.

Ethics declarations

Ethics approval and consent to participate

The study was approved by the Leeds East Research Ethics Committee (ethical approval 15/YH/0080) and performed in accordance with the Declaration of Helsinki. All samples were obtained with the patients’ informed consent.

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No individually identifiable data are presented.

Data availability

RNA-seq data, generated through our ICGC CAGEKID study, has been deposited in a public repository as described elsewhere: Scelo et al.30 Proteomic data sets are available on request.

Competing interests

A.H. is Editor-in-Chief of British Journal of Cancer. R.S.’s current role is Publishing Editor for British Journal of Cancer.

Funding information

This work was supported by funding from the University of Leeds and Cancer Research UK. E.R.M. is funded by the European Research Council (Advanced Researcher Award), NIHR (Senior Investigator Award and Cambridge NIHR Biomedical Research Centre) and Cancer Research UK Cambridge Cancer Centre. The views expressed are those of the authors and not necessarily those of the NHS or Department of Health. The University of Cambridge has received salary support in respect of E.R.M. from the NHS in the East of England through the Clinical Academic Reserve.

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Hornigold, N., Dunn, K.R., Craven, R.A. et al. Dysregulation at multiple points of the kynurenine pathway is a ubiquitous feature of renal cancer: implications for tumour immune evasion. Br J Cancer 123, 137–147 (2020). https://doi.org/10.1038/s41416-020-0874-y

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