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Cited by 3Cancer, Hypoxia, and MetabolismCancer, Lipids, and MetabolismMetabolism, Diabetes, and Cancer

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A Citrate Synthase Splice Variant Rewires the TCA Cycle to Promote Colorectal Cancer Progression

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Cheung, Justin Chak Ting, Ng, Lok Wan, Zhu, Zhongxu, Chen, Bonan, Li, Stephen, Xu, Mingjing, Ding, Xiaofan, Pu, Dandan, Hu, Yi, Ren, Yuqing, Kang, Wei, Li, Ming, Wong, Jason Wing Hon, Wang, Xin, Cheng, Yuen Kit, Aik, Wei Shen, Wong, Ka Leung, Ng, Simon Siu Man, Wong, Nathalie, Dong, Yujuan·2025·Cancer Research
DOI 10.1158/0008-5472.can-24-2355PMC PMC12616247

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Abstract

Metabolic reprogramming, notably alterations in the tricarboxylic acid (TCA) cycle, has emerged as a hallmark of cancer that supports tumor growth and metastasis. Despite the TCA cycle being a classical central metabolic pathway, further exploration is needed to fully elucidate the intricate manifestations and contributory mechanisms of TCA cycle rewiring in colorectal carcinogenesis. In this study, we identified a splicing isoform of citrate synthase (CS), CS-ΔEx4, and unveiled its role in TCA cycle dysregulation in colorectal cancer. CS-ΔEx4 was distinctly upregulated in colorectal cancer tumors compared with the canonical CS full-length (CS-FL) isoform. Clinical analyses established a strong correlation between elevated CS-ΔEx4 expression and cancer recurrence as well as inferior survival outcomes in patients with colorectal cancer. Functional experiments revealed the active contribution of CS-ΔEx4 to the aggressive phenotype of colorectal cancer cells both in vitro and in vivo. Mechanistically, CS-ΔEx4 formed a heterocomplex with CS-FL within the mitochondria that influenced the enzymatic function of canonical CS and accelerated TCA cycle flux, thereby promoting the accumulation of the oncometabolite 2-hydroxyglutarate. The CS-ΔEx4–mediated metabolic alterations engendered epigenomic modulations that drove the upregulation of oncogenic gene signatures. In silico screening identified a small molecule with potent antiproliferative effects in colorectal cancer cell line and organoid models that selectively antagonized the CS-ΔEx4 and CS-FL heterocomplex activity while sparing the CS-FL homodimers. Together, this study discovered the presence of a spliced CS isoform that promotes colorectal cancer progression and identified a molecule that holds potential for targeting the CS-ΔEx4 and CS-FL heterocomplex. Significance: The citrate synthase variant CS-ΔEx4 augments TCA metabolic flux to facilitate epigenetic reprogramming and colorectal carcinogenesis and can be targeted with a small molecule, providing a promising treatment strategy for colorectal cancer. See related commentary by Schatton and Frezza, p. 4293

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Metadata

Authors
Cheung, J. C. T., Ng, L. W., Zhu, Z., Chen, B., Li, S., Xu, M., Ding, X., Pu, D., Hu, Y., Ren, Y., Kang, W., Li, M., Wong, J. W. H., Wang, X., Cheng, Y. K., Aik, W. S., Wong, K. L., Ng, S. S. M., Wong, N., & Dong, Y.
Affiliations
  • Chinese University of Hong Kong (HK)
  • Hong Kong Baptist University (HK)
  • Chinese Academy of Sciences (CN)
  • University of Hong Kong (HK)
  • University of Macau (MO)
  • Nanchang University (CN)
  • Hong Kong Polytechnic University (HK)
Corresponding author
  • Wong, Nathalie
  • Dong, Yujuan
ORCID
Venue
Cancer Research
Publisher
American Association for Cancer Research (AACR)
Published
2025-09-03
Volume
85
Issue
22
Pages
4450-4468
ISSN
  • 0008-5472 (ISSN-L)
  • 1538-7445
DOI
10.1158/0008-5472.can-24-2355
PMC
PMC12616247
OpenAlex
W4413952524
Cited by
3
References
10
Citations by year
2026: 3
FWCI
1.35
Type
journal article
Language
en
Open access
Licence
Locations
  • Cancer Research (publishedVersion, cc-by-nc-nd)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • Research Grants Council, University Grants Committee (AoE/M-, 401/20, 14115123, C4024-22GF)
  • Innovation and Technology Fund (RAI/24/1/077A)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Lipids, and Metabolism
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolism, Diabetes, and Cancer
Keywords
citrate synthase, splice variants, colorectal cancer, 2-hydroxyglutarate, epigenetic reprogramming, tumor metabolic reprogramming, small molecule inhibitors, colorectal cancer progression, tumor recurrence
MeSH terms
Animals, Citrate (si)-Synthase, Citric Acid Cycle, Female, Humans, Mitochondria, Colorectal Neoplasms, Gene Expression Regulation, Neoplastic, Alternative Splicing, Disease Progression, Cell Line, Tumor, Cell Proliferation, Mice

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