Collagen Prolyl Hydroxylation–Dependent Metabolic Perturbation Governs Epigenetic Remodeling and Mesenchymal Transition in Pluripotent and Cancer Cells

Abstract

Collagen prolyl hydroxylation (CPH), which is catalyzed by prolyl 4-hydroxylase (P4H), is the most prevalent posttranslational modification in humans and requires vitamin C (VitC). Here, we demonstrate that CPH acts as an epigenetic modulator of cell plasticity. Increased CPH induced global DNA/histone methylation in pluripotent stem and tumor cells and promoted cell state transition (CST). Interfering with CPH by either genetic ablation of P4H subunit alpha-2 (P4HA2) or pharmacologic treatment reverted epigenetic changes and antagonized CST. Mechanistically, we suggest that CPH modifies the epigenetic landscape by reducing VitC for DNA and histone demethylases. Repurposed drugs targeting CPH-mediated metabolic perturbation, such as the antiasthmatic budesonide, blocked metastatic dissemination of breast cancer cells in vivo by preventing mesenchymal transition. Our study provides mechanistic insights into how metabolic cues and epigenetic factors integrate to control CST and paves the way for the development of novel antimetastatic strategies. Significance: A phenotype-based high-throughput screening reveals unforeseen metabolic control of cell plasticity and identifies budesonide as a drug candidate for metastatic cancer.

The role of collagen in cancer: from bench to bedside

“…Interfering collagen biosynthesis enzymes Collagen genes MiR-129-5p, MiR-29b, MiR-384 [275][276][277] Prolyl 4-hydroxylase Budesonide, catechol, N-oxalylglycine, coumalic acid, ethyl dihydroxybenzoic acid [278][279][280] Heat shock protein 90 1G6-D7, dipalmitoyl-radicicol, 17-DMAG, ganetespib [281][282][283][284] Heat shock protein 47 MiR-29, 1,3-dimethylol-5-FU, AK778, pirfenidone, terutroban [285,286] Matrix metalloproteinases Gallium complex GS2, isoflavonoids, bisphosphonates [287,288] Lysyl oxidases Beta-aminopropionitrile [289] Disturbing cancer cell signaling pathways Snail transcription factors Toosendanin, ponicidin, ferulic acid [290] Hypoxia-inducible factor Tamoxifen, 28-O-propynoylbetulin [291,292] STAT3 signaling pathway VS-4718, stattic, ruxolitinib, S3I-201 [293,294] TGF-β signaling pathway LY2157299 monohydrate, trabedersen, fresolimumab, galunisertib [295,296] NF-κB signaling pathway Honokiol, aspirin, ormeloxifene [297][298][299] AKT signaling pathway Quetiapine, pirfenidone [300] Notch signaling pathway Rovalpituzumab tesirine, taladegib, crenigacestat, MiR-148a [301] Hedgehog signaling pathway Itraconazole, sonidegib, vismodegib [302] RAS signaling pathway Perindopril, losartan [100,303] Tyrosine kinase receptor Bevacizumab, imatinib, ponatinib, dasatinib [304,305] Discoidin domain receptor WRG-28, 7rh, AZD0156 [306][307][308] G protein family receptor AT13148, KD025, Azaindole 1, chelerythrine…”

Proline Metabolism in Tumor Growth and Metastatic Progression

“…However, besides reduced proliferation/cell number and/or increased apoptosis, lower tumor volume can be the consequence of reduced stroma/ECM. Indeed, lower levels of Proline affect collagen/ECM accumulation, which eventually results in smaller/more compact tumors that have less capacity to invade and generate metastasis (57).…”

“…P4HA2 has been recently reported to play a role in the progression of breast ductal carcinoma in situ (DCIS) (67). Furthermore, high levels of P4HA2 has been associated with decreased survival in a breast cancer dataset with almost 2000 patients, and is an independent predictor of disease outcome with respect to standard clinopathological parameters (57). Accordingly, silencing of P4HA2 converted Triple Negative Breast Cancer (TNBC) cells to a more epithelial phenotype, and reduces invasiveness in a 3D organotypic culture (57).…”

“…Genetic mutations do not explain the heterogeneity/plasticity of cancer cells, which can be better explained by metabolic, epigenetic mechanisms. In this respect, we have recently proposed a novel mechanism underlying cancer cells plasticity by which collagen maturation may act as an epigenetic signal (57). Data provided in recently published papers underscore the existence of a functional link between Proline metabolism and epigenetic remodeling (84,96,97) and demonstrate that Proline availability influences mouse embryonic stem cell (mESC) identity and behavior through modulating AAR-ATF4 pathway (see above paragraph Proline availability controls cancer cell behavior).…”

“…Furthermore, esMT is accompanied by epigenetic remodeling, which results in a genome-wide increase of DNA and histone methylation (84,98). While several metabolites act as cofactors or substrates of epigenetic enzymes and may thus influence the epigenetic landscape (99), a different mechanism has been recently proposed underlying Proline's epigenetic activity, which relies on collagen synthesis/maturation that requires Vitamin C (VitC) (Figure 2) (57). Following this model, consequently to a rapid increase of Proline-dependent collagen synthesis, the activity of Prolyl-hydroxylase (P4h) enzymes for collagen maturation, consumes VitC in the ER.…”