Deciphering the mechanism of PSORI-CM02 in suppressing keratinocyte proliferation through the mTOR/HK2/glycolysis axis

Hyperplasia of epidermal keratinocytes that depend on glycolysis is a new hallmark of psoriasis pathogenesis. Our previous studies demonstrated that PSORI-CM02 could halt the pathological progression of psoriasis by targeting inflammatory response and angiogenesis, but its effect(s) and mechanism(s) on proliferating keratinocytes remained unclear. In this study, we aim to identify components of PSORI-CM02 that are absorbed into the blood and to determine the effect(s) of PSORI-CM02 on keratinocyte proliferation and its molecular mechanism(s). We used the immortalized human epidermal keratinocyte cell line, HaCaT, as an in vitro model of proliferating keratinocytes and the imiquimod-induced psoriasis mouse (IMQ) as an in vivo model. Metabolite profiles of vehicle pharmaceutic serum (VPS), PSORI-CM02 pharmaceutic serum (PPS), and water extraction (PWE) were compared, and 23 components of PSORI-CM02 were identified that were absorbed into the blood of mice. Both PPS and PWE inhibited the proliferation of HaCaT cells and consequently reduced the expression of the proliferation marker ki67. Additionally, PPS and PWE reduced phosphorylation levels of mTOR pathway kinases. Seahorse experiments demonstrated that PPS significantly inhibited glycolysis, glycolytic capacity, and mitochondrial respiration, thus reducing ATP production in HaCaT cells. Upon treatments of PPS or PWE, hexokinase 2 (HK2) expression was significantly decreased, as observed from the set of glycolytic genes we screened. Finally, in the IMQ model, we observed that treatment with PSORI-CM02 or BPTES, an inhibitor of mTOR signaling, reduced hyperproliferation of epidermal keratinocytes, inhibited the expression of p-S6 and reduced the number of proliferating cell nuclear antigen (PCNA)-positive cells in lesioned skin. Taken together, we demonstrate that PSORI-CM02 has an anti-proliferative effect on psoriatic keratinocytes, at least in part, by inhibiting the mTOR/HK2/glycolysis axis.

[1]  F. Baltazar,et al.  Targeting Monocarboxylate Transporters (MCTs) in cancer: How close are we to the clinics? , 2023, Seminars in cancer biology.

[2]  H. Jun,et al.  LPA/LPAR1 signaling induces PGAM1 expression via AKT/mTOR/HIF-1α pathway and increases aerobic glycolysis, contributing to keratinocyte proliferation. , 2022, Life sciences.

[3]  Hu Zhou,et al.  18F-FDG PET as an imaging biomarker for the response to FGFR-targeted therapy of cancer cells via FGFR-initiated mTOR/HK2 axis , 2022, Theranostics.

[4]  Haiyan Chen,et al.  Paeoniflorin Inhibits the Proliferation and Metastasis of Ulcerative Colitis-Associated Colon Cancer by Targeting EGFL7 , 2022, Journal of oncology.

[5]  R. Xu,et al.  Paeoniflorin Inhibits EMT and Angiogenesis in Human Glioblastoma via K63-Linked C-Met Polyubiquitination-Dependent Autophagic Degradation , 2022, Frontiers in Oncology.

[6]  S. Burley,et al.  mTOR regulates aerobic glycolysis through NEAT1 and nuclear paraspeckle-mediated mechanism in hepatocellular carcinoma , 2022, Theranostics.

[7]  Lian Cui,et al.  Advances in the pathogenesis of psoriasis: from keratinocyte perspective , 2022, Cell death & disease.

[8]  X. Dai,et al.  Pyruvate Kinase M2 Mediates Glycolysis Contributes to Psoriasis by Promoting Keratinocyte Proliferation , 2021, Frontiers in Pharmacology.

[9]  Yang Sun,et al.  Deciphering the mechanism of Fang-Ji-Di-Huang-Decoction in ameliorating psoriasis-like skin inflammation via the inhibition of IL-23/Th17 cell axis. , 2021, Journal of ethnopharmacology.

[10]  Weigang Zhang,et al.  Cyclin-Dependent Kinase 7 Promotes Th17/Th1 Cell Differentiation in Psoriasis by Modulating Glycolytic Metabolism. , 2021, The Journal of investigative dermatology.

[11]  Jing-jing Wu,et al.  Anti-Angiogenic Efficacy of PSORI-CM02 and the Associated Mechanism in Psoriasis In Vitro and In Vivo , 2021, Frontiers in Immunology.

[12]  Sung-Hoon Kim,et al.  Apoptotic effect of compound K in hepatocellular carcinoma cells via inhibition of glycolysis and Akt/mTOR/c‐Myc signaling , 2021, Phytotherapy research : PTR.

[13]  Ran Guo,et al.  Paeoniflorin Attenuates Dexamethasone-Induced Apoptosis of Osteoblast Cells and Promotes Bone Formation via Regulating AKT/mTOR/Autophagy Signaling Pathway , 2021, Evidence-based complementary and alternative medicine : eCAM.

[14]  J. Saraiva,et al.  Emergent Technologies for the Extraction of Antioxidants from Prickly Pear Peel and Their Antimicrobial Activity , 2021, Foods.

[15]  Ruifeng Liu,et al.  Dysregulated Dermal Mesenchymal Stem Cell Proliferation and Differentiation Interfered by Glucose Metabolism in Psoriasis , 2021, International journal of stem cells.

[16]  E. Jacinto,et al.  Regulation and metabolic functions of mTORC1 and mTORC2. , 2021, Physiological reviews.

[17]  L. Kuai,et al.  Uncovering the Mechanism of Jueyin Granules in the Treatment of Psoriasis Using Network Pharmacology. , 2020, Journal of ethnopharmacology.

[18]  E. Kontopantelis,et al.  National, regional, and worldwide epidemiology of psoriasis: systematic analysis and modelling study , 2020, BMJ.

[19]  A. Armstrong,et al.  Pathophysiology, Clinical Presentation, and Treatment of Psoriasis: A Review. , 2020, JAMA.

[20]  Zhirong Sun,et al.  Fasting inhibits aerobic glycolysis and proliferation in colorectal cancer via the Fdft1-mediated AKT/mTOR/HIF1α pathway suppression , 2020, Nature Communications.

[21]  Wei Sun,et al.  β-Asarone Increases Chemosensitivity by Inhibiting Tumor Glycolysis in Gastric Cancer , 2020, Evidence-based complementary and alternative medicine : eCAM.

[22]  Y. Lai,et al.  Keratinocyte: A trigger or an executor of psoriasis? , 2020, Journal of leukocyte biology.

[23]  Yanmei Zhang,et al.  Isorhamnetin inhibited the proliferation and metastasis of androgen-independent prostate cancer cells by targeting the mitochondrion-dependent intrinsic apoptotic and PI3K/Akt/mTOR pathway , 2020, Bioscience reports.

[24]  Hong-Yu Zhang,et al.  PSORI-CM02 formula alleviates imiquimod-induced psoriasis via affecting macrophage infiltration and polarization. , 2019, Life sciences.

[25]  S. Duan,et al.  mTOR‐mediated metabolic reprogramming shapes distinct microglia functions in response to lipopolysaccharide and ATP , 2019, Glia.

[26]  Han Ling,et al.  PSORI-CM02 ameliorates psoriasis in vivo and in vitro by inducing autophagy via inhibition of the PI3K/Akt/mTOR pathway. , 2019, Phytomedicine : international journal of phytotherapy and phytopharmacology.

[27]  A. Bode,et al.  Ethyl gallate as a novel ERK1/2 inhibitor suppresses patient‐derived esophageal tumor growth , 2019, Molecular carcinogenesis.

[28]  Wei Zhu,et al.  PSORI-CM02 alleviates IMQ-induced mouse dermatitis via differentially regulating pro- and anti-inflammatory cytokines targeting of Th2 specific transcript factor GATA3. , 2019, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[29]  R. Deberardinis,et al.  Differential Glucose Requirement in Skin Homeostasis and Injury Identifies a Therapeutic Target for Psoriasis , 2018, Nature Medicine.

[30]  Yongqi Fang,et al.  β‐asarone inhibited cell growth and promoted autophagy via P53/Bcl‐2/Bclin‐1 and P53/AMPK/mTOR pathways in Human Glioma U251 cells , 2018, Journal of cellular physiology.

[31]  Chuanjian Lu,et al.  PSORI-CM02 Formula Increases CD4+ Foxp3+ Regulatory T Cell Frequency and Ameliorates Imiquimod-Induced Psoriasis in Mice , 2018, Front. Immunol..

[32]  Serdal Kirmizialtin,et al.  The catalytic inactivation of the N-half of human hexokinase 2 and structural and biochemical characterization of its mitochondrial conformation , 2018, Bioscience reports.

[33]  W. Boehncke,et al.  Unmet Needs in the Field of Psoriasis: Pathogenesis and Treatment , 2018, Clinical Reviews in Allergy & Immunology.

[34]  P. Imbimbo,et al.  Protective effect of Opuntia ficus-indica L. cladodes against UVA-induced oxidative stress in normal human keratinocytes. , 2017, Bioorganic & medicinal chemistry letters.

[35]  L. Alvarez,et al.  Phenolic Compounds Isolated from Caesalpinia coriaria Induce S and G2/M Phase Cell Cycle Arrest Differentially and Trigger Cell Death by Interfering with Microtubule Dynamics in Cancer Cell Lines , 2017, Molecules.

[36]  S. Ki,et al.  Isorhamnetin Inhibits Reactive Oxygen Species-Dependent Hypoxia Inducible Factor (HIF)-1α Accumulation. , 2016, Biological & pharmaceutical bulletin.

[37]  S. Horvath,et al.  MCT1 Modulates Cancer Cell Pyruvate Export and Growth of Tumors that Co-express MCT1 and MCT4. , 2016, Cell reports.

[38]  Wei Zhang,et al.  Isorhamnetin inhibits cell proliferation and induces apoptosis in breast cancer via Akt and mitogen-activated protein kinase kinase signaling pathways , 2015, Molecular medicine reports.

[39]  M. Wang,et al.  Ethyl gallate suppresses proliferation and invasion in human breast cancer cells via Akt-NF-κB signaling. , 2015, Oncology reports.

[40]  Junbo Hu,et al.  Isorhamnetin suppresses colon cancer cell growth through the PI3K‑Akt‑mTOR pathway. , 2014, Molecular medicine reports.

[41]  Wei Wei,et al.  BAFF/BAFF-R involved in antibodies production of rats with collagen-induced arthritis via PI3K-Akt-mTOR signaling and the regulation of paeoniflorin. , 2012, Journal of ethnopharmacology.

[42]  E. Tisdale.  Glyceraldehyde-3-phosphate Dehydrogenase Is Phosphorylated by Protein Kinase Cι/λ and Plays a Role in Microtubule Dynamics in the Early Secretory Pathway* , 2002, The Journal of Biological Chemistry.

[43]  J. Miyazaki,et al.  Overexpression of hexokinase I but not GLUT1 glucose transporter alters concentration dependence of glucose-stimulated insulin secretion in pancreatic beta-cell line MIN6. , 1994, The Journal of biological chemistry.