Trypanosoma cruzi Dysregulates piRNAs Computationally Predicted to Target IL-6 Signaling Molecules During Early Infection of Primary Human Cardiac Fibroblasts

다국어 초록 (Multilingual Abstract)

Trypanosoma cruzi, the etiological agent of Chagas disease, is an intracellular protozoan parasite, which is now present in most industrialized countries. About 40% of T. cruzi infected individuals will develop severe, incurable cardiovascular, gastrointestinal, or neurological disorders. The molecular mechanisms by which T. cruzi induces cardiopathogenesis remain to be determined. Previous studies showed that increased IL-6 expression in T. cruzi patients was associated with disease severity. IL-6 signaling was suggested to induce pro-inflammatory and pro-fibrotic responses, however, the role of this pathway during early infection remains to be elucidated. We reported that T. cruzi can dysregulate the expression of host PIWI-interacting RNAs (piRNAs) during early infection. Here, we aim to evaluate the dysregulation of IL-6 signaling and the piRNAs computationally predicted to target IL-6 molecules during early T. cruzi infection of primary human cardiac fibroblasts (PHCF). Using in silico analysis, we predict that piR_004506, piR_001356, and piR_017716 target IL6 and SOCS3 genes, respectively. We validated the piRNAs and target gene expression in T. cruzi challenged PHCF. Secreted IL-6, soluble gp-130, and sIL-6R in condition media were measured using a cytokine array and western blot analysis was used to measure pathway activation. We created a network of piRNAs, target genes, and genes within one degree of biological interaction. Our analysis revealed an inverse relationship between piRNA expression and the target transcripts during early infection, denoting the IL-6 pathway targeting piRNAs can be developed as potential therapeutics to mitigate T. cruzi cardiomyopathies.

참고문헌 (Reference)

1 Simonelig M, "piRNAs, master regulators of gene expression" 24 : 779-780, 2014

2 Rayford KJ, "piRNAs as modulators of disease pathogenesis" 22 : 2373-, 2021

3 Weick EM, "piRNAs : from biogenesis to function" 141 : 3458-3471, 2014

4 Laugier L, "miRNAs may play a major role in the control of gene expression in key pathobiological processes in Chagas disease cardiomyopathy" 14 : e0008889-, 2020

5 Cheng ZY, "ZBTB transcription factors : key regulators of the development, differentiation and effector function of T cells" 12 : 713294-, 2021

6 Martinez VD, "Unique somatic and malignant expression patterns implicate PIWI-interacting RNAs in cancer-type specific biology" 5 : 10423-, 2015

7 Ponce NE, "Trypanosoma cruzi, the causative agent of Chagas disease, modulates interleukin-6-induced STAT3 phosphorylation via gp130 cleavage in different host cells" 1832 : 485-494, 2013

8 Nisimura LM, "Trypanosoma cruzi promotes transcriptomic remodeling of the JAK/STAT signaling and cell cycle pathways in myoblasts" 10 : 255-, 2020

9 Stahl P, "Trypanosoma cruzi parasites fight for control of the JAK-STAT pathway by disarming their host" 3 : e1012964-, 2015

10 Rayford KJ, "Trypanosoma cruzi modulates PIWI-interacting RNA expression in primary human cardiac myocytes during the early phase of infection" 21 : E9439-, 2020

11 Tshori S, "Transcription factor MITF regulates cardiac growth and hypertrophy" 116 : 2673-2681, 2006

12 Ponce NE, "Toll-like receptor-2 and interleukin-6 mediate cardiomyocyte protection from apoptosis during Trypanosoma cruzi murine infection" 201 : 145-155, 2012

13 Arun A, "Thrombospondin-1 expression and modulation of Wnt and hippo signaling pathways during the early phase of Trypanosoma cruzi infection of heart endothelial cells" 16 : e0010074-, 2022

14 Müllberg J, "The soluble interleukin-6 receptor is generated by shedding" 23 : 473-480, 1993

15 Jones SA, "The role of soluble receptors in cytokine biology : the agonistic properties of the sIL-6R/IL-6 complex" 1592 : 251-263, 2002

16 Hinz B, "The role of myofibroblasts in wound healing" 64 : 171-177, 2016

17 Johnson BZ, "The role of IL-6 in skin fibrosis and cutaneous wound healing" 8 : 101-, 2020

18 Galun E, "The regenerative activity of interleukin-6" 982 : 59-77, 2013

19 Klingberg F, "The myofibroblast matrix : implications for tissue repair and fibrosis" 229 : 298-309, 2013

20 Warde-Farley D, "The GeneMANIA prediction server : biological network integration for gene prioritization and predicting gene function" 38 : W214-W220, 2010

21 McGeachy MJ, "TGF-beta and IL-6 drive the production of IL-17 and IL-10 by T cells and restrain T(H)-17 cell-mediated pathology" 8 : 1390-1397, 2007

22 Hoffman KA, "Signal transducer and activator of transcription-3 modulation of cardiac pathology in chronic Chagasic cardiomyopathy" 11 : 708325-, 2021

23 Kulkarni MM, "Signal transducer and activator of transcription 1(STAT-1)plays a critical role in control of Trypanosoma cruzi infection" 145 : 225-231, 2015

24 Schumacher N, "Shedding of endogenous interleukin-6 receptor(IL-6R)is governed by a disintegrin and metalloproteinase(ADAM)proteases while a full-length IL-6R isoform localizes to circulating microvesicles" 290 : 26059-26071, 2015

25 Volta BJ, "Serum cytokines as biomarkers of early Trypanosoma cruzi infection by congenital exposure" 196 : 4596-4602, 2016

26 Zhu BM, "SOCS3 negatively regulates the gp130-STAT3 pathway in mouse skin wound healing" 128 : 1821-1829, 2008

27 Waghabi MC, "SB-431542, a transforming growth factor beta inhibitor, impairs Trypanosoma cruzi infection in cardiomyocytes and parasite cycle completion" 51 : 2905-2910, 2007

28 Fredj S, "Role of interleukin-6 in cardiomyocyte/cardiac fibroblast interactions during myocyte hypertrophy and fibroblast proliferation" 204 : 428-436, 2005

29 Yamada D, "Role of crosstalk between interleukin-6 and transforming growth factor-beta 1 in epithelial-mesenchymal transition and chemoresistance in biliary tract cancer" 49 : 1725-1740, 2013

30 Reeh H, "Response to IL-6 trans-and IL-6 classic signalling is determined by the ratio of the IL-6 receptor α to gp130 expression : fusing experimental insights and dynamic modelling" 17 : 46-, 2019

31 Ferreira JC, "Regulation of cardiac excitability by protein kinase C isozymes" 4 : 532-546, 2012

32 Watanabe T, "Posttranscriptional regulation of gene expression by Piwi proteins and piRNAs" 56 : 18-27, 2014

33 Peng CY, "Positive feedback loop of Snail-IL-6 mediates myofibroblastic differentiation activity in precancerous oral submucous fibrosis" 12 : E1611-, 2020

34 Murakami M, "Pleiotropy and specificity : insights from the interleukin 6 family of cytokines" 50 : 812-831, 2019

35 Suman S, "Phospho-proteomic analysis of primary human colon epithelial cells during the early Trypanosoma cruzi infection phase" 12 : e0006792-, 2018

36 Antunes D, "Oral route driven acute Trypanosoma cruzi infection unravels an IL-6 dependent hemostatic derangement" 10 : 1073-, 2019

37 Braun MU, "Opposing effects of delta-and zeta-protein kinase C isozymes on cardiac fibroblast proliferation : use of isozyme-selective inhibitors" 35 : 895-903, 2003

38 Schneider CA, "NIH image to ImageJ : 25 years of image analysis" 9 : 671-675, 2012

39 Rodríguez-Angulo HO, "Molecular remodeling of cardiac sinus node associated with acute Chagas disease myocarditis" 9 : 2208-, 2021

40 Navarro IC, "MicroRNA transcriptome profiling in heart of Trypanosoma cruzi-infected mice : parasitological and cardiological outcomes" 9 : e0003828-, 2015

41 Ma F, "Macrophage-stimulated cardiac fibroblast production of IL-6 is essential for TGF β/Smad activation and cardiac fibrosis induced by angiotensin II" 7 : e35144-, 2012

42 Gebert D, "Large Drosophila germline piRNA clusters are evolutionarily labile and dispensable for transposon regulation" 81 : 3965-3978, 2021

43 Gotthardt D, "JAK/STAT cytokine signaling at the crossroad of NK cell development and maturation" 10 : 2590-, 2019

44 O’Reilly S, "Interleukin-6(IL-6)trans signaling drives a STAT3-dependent pathway that leads to hyperactive transforming growth factor-β(TGF-β)signaling promoting SMAD3 activation and fibrosis via Gremlin protein" 289 : 9952-9960, 2014

45 Truyens C, "Interleukin-6(IL-6)production in mice infected with Trypanosoma cruzi : effect of its paradoxical increase by anti-IL-6 monoclonal antibody treatment on infection and acute-phase and humoral immune responses" 62 : 692-696, 1994

46 Fielding CA, "Interleukin-6 signaling drives fibrosis in unresolved inflammation" 40 : 40-50, 2014

47 Zhang XL, "Interleukin-6 regulation of transforming growth factor(TGF)-beta receptor compartmentalization and turnover enhances TGF-beta1 signaling" 280 : 12239-12245, 2005

48 Chi L, "Interleukin-6 production by endothelial cells via stimulation of protease-activated receptors is amplified by endotoxin and tumor necrosis factor-alpha" 21 : 231-240, 2001

49 Gabay C, "Interleukin-6 and chronic inflammation" 8 (8): S3-, 2006

50 Ferreira LR, "Integration of miRNA and gene expression profiles suggest a role for miRNAs in the pathobiological processes of acute Trypanosoma cruzi infection" 7 : 17990-, 2017

51 Herrera RN, "Inflammatory and prothrombotic activation with conserved endothelial function in patients with chronic, asymptomatic Chagas disease" 17 : 502-507, 2011

52 Hinterseher I, "Immunohistochemical analysis of the natural killer cell cytotoxicity pathway in human abdominal aortic aneurysms" 16 : 11196-11212, 2015

53 Hirano T, "IL-6 in inflammation, autoimmunity and cancer" 33 : 127-148, 2021

54 Gubernatorova EO, "IL-6 : relevance for immunopathology of SARS-CoV-2" 53 : 13-24, 2020

55 Kishimoto T, "IL-6 : from its discovery to clinical applications" 22 : 347-352, 2010

56 Cevey AC, "IL-10/STAT3/SOCS3 axis is involved in the anti-inflammatory effect of benznidazole" 10 : 1267-, 2019

57 Kumar S, "HIMF(hypoxia-induced mitogenic factor)-IL(interleukin)-6 signaling mediates cardiomyocyte-fibroblast crosstalk to promote cardiac hypertrophy and fibrosis" 73 : 1058-1070, 2019

58 Hershey CL, "Genomic analysis of the Microphthalmia locus and identification of the MITF-J/Mitf-J isoform" 347 : 73-82, 2005

59 Oliveira AER, "Gene expression network analyses during infection with virulent and avirulent Trypanosoma cruzi strains unveil a role for fibroblasts in neutrophil recruitment and activation" 16 : e1008781-, 2020

60 Ghansah TJ, "Epidermal growth factor binds to a receptor on Trypanosoma cruzi amastigotes inducing signal transduction events and cell proliferation" 49 : 383-390, 2002

61 Olivera MJ, "Economic costs of Chagas disease in Colombia in 2017 : a social perspective" 91 : 196-201, 2020

62 Udoko AN, "Early regulation of profibrotic genes in primary human cardiac myocytes by Trypanosoma cruzi" 10 : e0003747-, 2016

63 Rodríguez-Angulo H, "Differential cytokine profiling in Chagasic patients according to their arrhythmogenic-status" 17 : 221-, 2017

64 Thiruvenkatarajan V, "Comment on Hamblin et al. Capillary ketone concentrations at the time of colonoscopy : a cross-sectional study with implications for SGLT2 inhibitor-treated type 2 diabetes. Diabetes Care 2021;44 : e124-e126" 45 : e15-e16, 2022

65 Abuhab A, "Chagas’ cardiomyopathy : the economic burden of an expensive and neglected disease" 168 : 2375-2380, 2013

66 Lidani KC, "Chagas disease : from discovery to a worldwide health problem" 7 : 166-, 2019

67 Nunes MCP, "Chagas cardiomyopathy : an update of current clinical knowledge and management : a scientific statement from the American Heart Association" 138 : e169-e209, 2018

68 Lai YH, "Cardiac remodeling, adaptations and associated myocardial mechanics in hypertensive heart diseases" 29 : 64-70, 2013

69 López L, "C-reactive protein and interleukin-6 serum levels increase as Chagas disease progresses towards cardiac failure" 59 : 50-56, 2006

70 Yue B, "Biology of the extracellular matrix : an overview" 23 : S20-S23, 2014

71 Nguyen HN, "Autocrine loop involving IL-6 family member LIF, LIF receptor, and STAT4 drives sustained fibroblast production of inflammatory mediators" 46 : 220-232, 2017

72 Waghabi MC, "Activation of transforming growth factor beta by Trypanosoma cruzi" 7 : 511-517, 2005

73 Niemand C, "Activation of STAT3 by IL-6 and IL-10 in primary human macrophages is differentially modulated by suppressor of cytokine signaling 3" 170 : 3263-3272, 2003

74 Yajima T, "Absence of SOCS3 in the cardiomyocyte increases mortality in a gp130-dependent manner accompanied by contractile dysfunction and ventricular arrhythmias" 124 : 2690-2701, 2011

75 Gomes Dos Santos A, "A specific IL6 polymorphic genotype modulates the risk of Trypanosoma cruzi parasitemia while IL18, IL17A, and IL1B variant profiles and HIV infection protect against cardiomyopathy in Chagas disease" 11 : 521409-, 2020

76 Jones BC, "A somatic piRNA pathway in the Drosophila fat body ensures metabolic homeostasis and normal lifespan" 7 : 13856-, 2016

77 Huang S, "A review of discovery profiling of PIWI-interacting RNAs and their diverse functions in metazoans" 22 : 11166-, 2021

78 Girard A, "A germline-specific class of small RNAs binds mammalian PIWI proteins" 442 : 199-202, 2006

79 Zhong F, "A SnoRNA-derived piRNA interacts with human interleukin-4 pre-mRNA and induces its decay in nuclear exosomes" 43 : 10474-10491, 2015