Localisation of PGK1 determines metabolic phenotype to balance metastasis and proliferation in patients with SMAD4-negative pancreatic cancer

Abstract

ObjectivePancreatic ductal adenocarcinoma (PDAC) is the most aggressive type of GI tumour, and it possesses deregulated cellular energetics. Although recent advances in PDAC biology have led to the discovery of recurrent genetic mutations in Kras, TP53 and SMAD4, which are related to this disease, clinical application of the molecular phenotype of PDAC remains challenging.DesignWe combined molecular imaging technology (positron emission tomography/CT) and immunohistochemistry to evaluate the correlation between the maximum standardised uptake value and SMAD4 expression and examined the effect of SMAD4 on glycolysis through in vitro and in vivo experiments. Furthermore, we identified the effect of SMAD4 on metabolic reprogramming by metabolomics and glucose metabolism gene expression analyses. Dual luciferase reporter assays and chromatin immunoprecipitation were performed to identify whether SMAD4 functioned as a transcription factor for phosphoglycerate kinase 1 (PGK1) in PDAC cells. Proliferative and metastatic assays were performed to examine the effect of PGK1 on the malignant behaviour of PDAC.ResultsWe provide compelling evidence that the glycolytic enzyme PGK1 is repressed by transforming growth factor-β/SMAD4. Loss of SMAD4 induces PGK1 upregulation in PDAC, which enhances glycolysis and aggressive tumour behaviour. Notably, in SMAD4-negative PDAC, nuclear PGK1 preferentially drives cell metastasis via mitochondrial oxidative phosphorylation induction, whereas cytoplasmic PGK1 preferentially supports proliferation by functioning as a glycolytic enzyme. The PDAC progression pattern and distinct PGK1 localisation combine to predict overall survival and disease-free survival.ConclusionPGK1 is a decisive oncogene in patients with SMAD4-negative PDAC and can be a target for the development of a therapeutic strategy for SMAD4-negative PDAC.

Precise and efficient silencing of mutant KrasG12D by CRISPR-CasRx controls pancreatic cancer progression

“…The AAV packaging was conducted as previously described 33 . Briefly, recombinant AAV (rAAV) vectors were generated by the transient transfection of 293T cells using three AAV plasmids (pAAV8-rep/cap-Y-F mutant, pAAV2-NLS-CasRx-NLS-HA-gRNA/pAAV2-GFP, and pHelper).…”

“…PDAC PDX mouse models were established by using freshly isolated pancreatectomy samples as previously described 33 . Briefly, every PDAC sample was isolated in two parts.…”

Molecular alterations and targeted therapy in pancreatic ductal adenocarcinoma

“…DSBs are repaired by BRCA through the gene conversion (GC) pathway in normal cells. However, in BRCA-loss cancer cells, DSBs cannot be repaired and will lead to fatal genomic instability metastatic patterns of PDAC cells and providing guidance for precise therapy [130].…”

Regulation of metabolic reprogramming by tumor suppressor genes in pancreatic cancer

“…SMAD4 can also function as a negative regulatory element in the glucose transport system, targeted by specific miRNAs, such as miR-29a and miR-23a [45]. Liang et al found loss of SMAD4 can enhance glycolysis and aggressive tumor behavior by upregulating phosphoglycerate kinase 1 (PGK1) in PDAC [19]. The relationship between SMAD4 and glutamine metabolism in tumors remains to be further studied.…”

“…All clinical trials failed so far either due to its negative safety profiles or the slight effect in clinical trials which is significantly attenuated compared with the effects in preclinical trials [15][16][17]. While on the other hand, more and more research indicated that a series of tumor suppressor genes may provide new ideas in effective pancreatic cancer treatments [18][19][20][21]. Therefore, in this paper, we reviewed recent research about metabolic reprogramming by tumor suppressor genes in pancreatic cancer, and hope to identify novel molecular targets for the development of chemotherapeutic approaches in PDAC (pancreatic ductal adenocarcinoma).…”