PFKFB3 overexpression in monocytes of patients with colon but not rectal cancer programs pro-tumor macrophages and is indicative for higher risk of tumor relapse

Abstract

IntroductionCirculating monocytes are main source for tumor-associated macrophages (TAMs) that control tumor growth, angiogenesis, metastasis and therapy resistance. We raised the questions how monocyte programming is affected by growing tumors localized in colon and rectal sections, and how treatment onsets affect monocyte programming in the circulation.MethodsPatients with rectal cancer and colon cancer were enrolled in the study. Peripheral blood monocytes were characterized by phenotypic analysis using flow cytometry, by transcriptomic analysis using RNA sequencing and by gene expression analysis using real-time RT-PCR. Phenotypic analysis was performed with IF/confocal microscopy. Spatial transcriptomic analysis was applied using GeoMX DSP-NGS.ResultsIn patients with rectal cancer, increased amount of CCR2+ monocytes was indicative for the absence of both lymphatic and hematogenous metastasis. In contrast, in patients with colon cancer CD163+ monocytes were indicative for LN metastasis. NGS analysis identified tumor-specific transcriptional programming of monocytes in all CRC patients compared to healthy individuals. The key transcriptional difference between monocytes of patients with colon and rectal cancer was increased expression of PFKFB3, activator of glycolysis that is currently considered as therapy target for major solid cancers. PFKFB3-expressing monocyte-derived macrophages massively infiltrated tumor in colon. Nanostring technology identified correlation of PFKFB3 with amount and tumor-promoting properties of TAMs in colon but not in rectal cancer. PFKFB3 was indicative for tumor relapse specifically in colon cancer.DiscussionOur findings provide essential argument towards CRC definition to cover two clinically distinct cancers – colon cancer and rectal cancer, that differentially interact with innate immunity.

Specific changes in amino acid profiles in monocytes of patients with breast, lung, colorectal and ovarian cancers

“…Two millions of monocytes were isolated from 10 ml of blood of a patient. The isolation of monocytes from peripheral blood was performed using density gradients followed by positive magnetic selection with CD14+ MACS beads (#130-050-201, Miltenyi Biotech, Germany) as previously described ( 30 ). The isolation quality of monocytes was determined by flow cytometry.…”

“…Intervening in the pathways and mechanisms involved in monocyte and TAMs reprogramming holds promising potential as a novel therapeutic strategy for personalized cancer immunotherapy ( 14 ). We and others have previously identified that monocytes undergo cancer-specific programming that can be detected on the level of phenotype (surface markers) or on the level transcriptome ( 3 , 30 , 31 ). In breast cancer, CD14 low CD16+HLA-DR+ monocytes were indicative for the good response to neoadjuvant chemotherapy ( 3 ).…”

“…In breast cancer, CD14 low CD16+HLA-DR+ monocytes were indicative for the good response to neoadjuvant chemotherapy ( 3 ). In colon, but not in rectal cancer, overexpression of activator of glycolysis PFKFB3 in monocytes, was correlated with tumor elimination after therapy ( 30 ). Metabolism of immune cells is critical for the ability to stimulate or suppress tumor growth.…”

The macrophage-associated prognostic gene ANXA5 promotes immunotherapy resistance in gastric cancer through angiogenesis

“…These immune constituents play pivotal roles in tumor invasion, metastasis, and clinical outcomes [ 6 , 7 ]. Specifically, tumor-associated macrophages (TAMs) serve as central innate immune components, facilitating both tumor growth and dissemination [ 8 ]. TAMs exhibit dual functions, promoting tumor growth through the M2 phenotype and suppressing it via the M1 phenotype [ 9 ].…”

Commentary: PFKFB3 overexpression in monocytes of patients with colon but not rectal cancer programs pro-tumor macrophages and is indicative for higher risk of tumor relapse

“…Larionova et al. have convincingly illustrated that monocytes, isolated from the peripheral blood of patients with colon cancer, demonstrate pronounced upregulation of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3), a key enzyme in the glycolytic cascade ( 1 ). In our assessment, this seminal work paves the way for extensive future research into immunogenesis and inflammation in colon cancer.…”