PFKFB4 is overexpressed in clear-cell renal cell carcinoma promoting pentose phosphate pathway that mediates Sunitinib resistance

Abstract

Background Kinases play critical role in clear-cell renal cell carcinoma (ccRCC). We aim to exploit novel kinase that is both protumorigenic and drugable in ccRCC. Methods Reproduction of public datasets with validation using microarray was performed to identify candidate gene. Functionality was studied using multi-omics with validation in vitro and in vivo. Results 6-Phosphofructo-2-Kinase/Fructose-2,6-Biphosphatase 4 (PFKFB4) was differentially expressed showing significantly higher expression in tumor than in normal kidney. PFKFB4 overexpression was associated with advanced tumor grade, stage and worsened prognosis. PFKFB4-knockdown significantly impaired fitness in cell proliferation, migration and wound healing. Despite being recurrently deleted on 3p, PFKFN4 mRNA remained actively transcribed by HIF1α. Metabolomics showed overexpressed PFKFB4 showed enriched metabolites in pentose phosphate pathway (PPP). Phosphoproteomics and immunoprecipitation showed PFKFB4 also phosphorylated NCOA3 which interacted with FBP1 to counteract overactive PPP flux, forming a regulatory loop. PFKFB4-knockdown overcame resistance to Sunitinib in vitro and in vivo both in xenograft and tail-vein injection murine models. Conclusion We concluded PFKFB4 was associated with PPP activity and the fine-tuning of which was mediated by its phosphorylation of NCOA3. Targeting PFKFB4 held promise to combat resistance to Sunitinib.

Current Status of the Use of Multifunctional Enzymes as Anti-Cancer Drug Targets

“…Although the great majority of the studies were focused on the inhibition of PFKFB3, there is record of at least one promising PFKFB4 inhibitor, 5-(n-(8-methoxy-4-quinolyl)amino)pentyl nitrate (5MPN) (Figure 4), that demonstrated an anti-proliferative effect of cancer cells both in vitro as in vivo [51]; 5MPN is a PFKFB4 specific inhibitor that binds competitively to its F6P binding site, suppressing kinase activity and consequently reducing the intracellular concentration of F-2,6-BP [51]. Recently, 5MPN was tested in combination therapy with Sunitinib, a receptor tyrosine kinase inhibitor used as a chemotherapeutic agent, showing promising results [52].…”

Construction and Validation of a Necroptosis-Related Signature Associated With the Immune Microenvironment in Liver Hepatocellular Carcinoma

“…Eight genes (PFKFB4, MSC-AS1, SPESP1, CTSV, CHST1, TMCO3, BMP6, and PAGE2) were incorporated into the final prognostic signature. It is reported that 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4 (PFKFB4) is a metabolic enzyme that mediates the proliferation, progression, and drug resistance of a variety of tumors through glucose metabolism ( Dasgupta et al, 2018 ; Gao et al, 2018 ; Feng et al, 2021 ; Shen et al, 2021 ). Musculin antisense RNA 1 (MSC-AS1) is a lncRNA, which interacts largely with miRNAs and acts as an oncogene in several cancers ( Li et al, 2021 ; Liu et al, 2021 ; Ma et al, 2021 ; Zhao et al, 2021 ).…”

Effect of PFKFB4 on the Prognosis and Immune Regulation of NSCLC and Its Mechanism

“…Warburg pathway enzyme fructose-6-phosphate 2-kinase/fructose-2,6-bisphosphatase 4 (PFKFB4) is a newly identified key factor in the regulation of transcriptional reprogramming. 7 PFKFB4 has been reported to be overexpressed in gastric, breast, thyroid, bladder, and colon cancer tissues; however, relatively little is known about the function and biological role of PFKFB4 in NSCLC.…”