Klotho expression in peripheral blood circulating cells is associated with vascular and systemic inflammation in atherosclerotic vascular disease

Abstract

Cardiovascular disease is the leading cause of death worldwide. New therapeutic strategies are aimed to modulate the athero-inflammatory process that partially orchestrates underlying vascular damage. Peripheral blood circulating cells include different immune cells with a central role in the development of the atherogenic inflammatory response. The anti-aging protein α-Klotho has been related to protective effects against CVD. KL is expressed in monocytes, macrophages, and lymphocytes where it exerts anti-inflammatory effects. In this work, we analyse the relationships of the levels of inflammatory markers with the expression of the KL gene in PBCCs and with the serum levels of soluble KL in atherosclerotic vascular disease. For this, we conducted a cross-sectional single-center case–control study including a study group of 76 CVD patients and a control group of 16 cadaveric organ donors without medical antecedent or study indicating CVD. Vascular artery fragments and whole blood and serum samples were obtained during elective or organ retrieval surgery. Serum levels of sKL, TNFα and IL10, and gene expression levels of KL, TNF, IL10, NFKB1, DNMT1, and DNMT3A in PBCCs were measured. In these cells, we also determined KL promoter methylation percentage. Histological and immunohistochemical analyses were employed to visualize atherosclerotic lesions and to measure IL10 and TNFα levels in vascular fragments. Patients with CVD presented higher values of proinflammatory markers both at systemic and in the vasculature and in the PBCCs, compared to the control group. In PBCCs, CVD patients also presented lower gene expression levels of KL gene (56.4% difference, P < 0.001), higher gene expression levels of DNMT1 and DNMT3A (P < 0.0001, for both) and a higher methylation status of in the promoter region of KL (34.1 ± 4.1% vs. 14.6 ± 3.4%, P < 0.01). In PBCCs and vasculature, KL gene expression correlated inversely with pro-inflammatory markers and directly with anti-inflammatory markers. sKL serum levels presented similar associations with the expression levels of pro- and anti-inflammatory markers in PBCCs. The differences in KL expression levels in PBCCs and in serum sKL levels with respect to control group was even greater in those CVD patients with macroscopically observable atheromatous plaques. We conclude that promoter methylation-mediated downregulation of KL gene expression in PBCCs is associated with the pro-inflammatory status in atherosclerotic vascular disease.

Screening and identification of key biomarkers associated with endometriosis using bioinformatics and next generation sequencing data analysis

“…ROBO2 [760], VCAM1 [761], GRP (gastrin releasing peptide) [762], FABP4 [763], ANO1 [764], SOX6 [765], TFAP2C [766], RAMP3 [767], PLA2G7 [768], MMP12 [769], FAIM2 [770], APOD (apolipoprotein D) [771], LAG3 [772], SOX18 [773], F2RL2 [774], CCR1 [775], FLT1 [776], FABP5 [629], TRPC3 [777]. THSD7A [778], DKK2 [779], PRKCB (protein kinase C beta) [780], DHCR24 [781], PDE3B [782], BMP4 [783], IL1RL1 [784], MYPN (myopalladin) [785], PLCG2 [786], PRL (prolactin) [787], WNT5A [788], MEOX1 [789], TIMP3 [790], FRZB (frizzled related protein) [791], CPE (carboxypeptidase E) [792], ADAMTS9 [793], NDNF (neuron derived neurotrophic factor) [794], PDGFB (platelet derived growth factor subunit B) [795], PIK3CG [796], LDLR (low density lipoprotein receptor) [797], CD4 [798], TRPA1 [799], F2RL3 [800], C1QL1 [801], ADAMTS5 [802], PDE4B [803], NES (nestin) [804], TH (tyrosine hydroxylase) [805], MMP8 [806], KDR (kinase insert domain receptor) [807], ADRB2 [808], ACKR3 [809], PTPRC (protein tyrosine phosphatase receptor type C) [810], KL (klotho) [811], KL (klotho) [812], PLAU (plasminogen activator, urokinase) [813], CCND2 [814], PTGS1 [815], INSIG1 [816], IRX2 [817], SIGLEC1 [818], UCN2 [819], CYP2J2 [820], CYP1A1 [821], ASTN2 [822], NTN1 [823], PDGFD (platelet derived growth factor D) [824], MSTN (myostatin) [663], LEPR (leptin receptor) [664], IL15 [825], CACNA1H [826], BMP2 [827], SYT7 [<...…”

Pathobiology of the Klotho Antiaging Protein and Therapeutic Considerations

“…Klotho is primarily produced in the kidney (renal tubules), but is also found in the brain (choroid plexus, CSF and neurons), pancreatic β cells, blood vessels and skin (Lim K, et al, 2015). Recent work also documents expression in peripheral blood circulating cells ( Martin-Nunez et al, 2022 ). It is also excreted into the urine.…”

“…Kidneys: Cheng et al, 2010 ; Kuro-o, 2010 , 2019 , 2021 ; Maquigussa et al, 2018 ; Oishi et al, 2021 ; Prud’homme et al, 2017a , 2020 ; Saar-Kovrov et al, 2021 ; Takenaka et al, 2017 ; Xing et al, 2021 ; Zou et al, 2018 . Cardiovascular: Chen K, et al, 2021 ; Freunlich et al, 2021 ; Manrique et al, 2016 ; Myung et al, 2022 ; Martin-Nunez et al, 2022 ; Olejnik et al, 2018 , 2020 ; Tyrenkov et al, 2021 ; Wang F and Zheng, 2022 ; Yang et al, 2015 . Brain: Dias et al, 2021 ; Driscoll et al, 2021 ; Cararo-Lopes et al, 2017 ; Fung et al, 2022 ; Hanson et al, 2021 ; Kundu et al, 2022 ; Laszczyk et al, 2017 ; Mytych, 2022 ; Nietzel et al, 2021; Sanz et al, 2021 .…”

Functional and Extracellular Production and Antitumor Activity of Mouse Alpha Klotho in Model Microalga Chlamydomonas reinhardtii

“…[18] Furthermore, sKL has been shown to have immunomodulatory effects, which means that it can help regulate the immune system and improve immune function. [19,20] Microalgae stands as a promising biological platform for the cost-effective and sustainable production of recombinant proteins and bioproducts using only water, CO 2, and sunlight. The microalgae C. reinhardtii is one of the most studied algae species and has been developed as a potent and photosynthetic expression system for the production of recombinant proteins, antigens, fatty acids, and chemicals with wide industrial applications.…”

“…In addition, the restoration of low sKL levels in cancer patients is eligible to alleviate cancer development in those immunocompromised patients with weakened immune systems [18] . Furthermore, sKL has been shown to have immunomodulatory effects, which means that it can help regulate the immune system and improve immune function [19,20] …”