Ignite Creation Date:
2024-05-06 @ 7:59 PM
Last Modification Date:
2024-10-26 @ 3:18 PM
Study NCT ID:
NCT06213688
Status:
NOT_YET_RECRUITING
Last Update Posted:
2024-01-19
First Post:
2024-01-10
Brief Title:
Involvement of Pollutants in Atopic Dermatitis and Psoriasis
Sponsor:
Assistance Publique Hopitaux De Marseille
Organization:
Assistance Publique Hopitaux De Marseille
Study Overview
Official Title:
The Impact of Pollutants on Patients With Psoriasis or Atopic Dermatitis by Investigating the Activation of the Aromatic Hydrocarbon Receptor
Status:
NOT_YET_RECRUITING
Status Verified Date:
2024-01
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
DIAhR
Brief Summary:
Introduction Pollution is a significant public health issue Research has shown a positive correlation between air pollution and chronic inflammatory dermatoses including psoriasis and eczema The incidence of these diseases has been steadily increasing since the beginning of industrialization The mechanism behind this association involves the activation of the aromatic hydrocarbon receptor AhR The aryl hydrocarbon receptor AhR plays a role in regulating the balance between T helper 17 TH17 and regulatory T cells TREG as well as in generating oxidative stress and producing pro-inflammatory cytokines Studies in cultured keratinocytes have shown that a non-competitive antagonist that modulates AhR activity can reduce cutaneous inflammatory processes induced by polycyclic aromatic hydrocarbons PAHs
Objectives
It has been suggested that activation of the AhR by PAHs and dioxins may be related to the pathogenesis of atopic dermatitis and psoriasis The main objective is to compare the levels of AhR pathway activation markers between cases and controls Secondary objectives include correlating environmental exposure to AhR ligands with disease severity in patients Finally we will compare the expression of inflammatory and AhR activation markers in cultured peripheral blood mononuclear cells PBMCs after in vitro stimulation with benzoapyrene
Material and methods
The study will measure exposure to pollutants by determining blood dioxins and urinary PAH metabolites Pro-inflammatory cytokines IL1β TNFα IL23 IL17 and IFNγ and Malondialdehyde MDA serum concentrations will be measured by ELISA The TREG and TH17 lymphocyte population ratio will be evaluated by flow cytometry on isolated PBMCs Additionally the level of expression of CYP 1A1 and 1B1 pollutant-metabolizing enzymes induced by AhR will be assessed on isolated PBMCs The expression levels of the AhR and NfkB active fractions will be determined by immunofluorescence Subsequently levels of AhR activation markers will be compared after stimulation of PBMCs with benzoapyrene
Objectives
It has been suggested that activation of the AhR by PAHs and dioxins may be related to the pathogenesis of atopic dermatitis and psoriasis The main objective is to compare the levels of AhR pathway activation markers between cases and controls Secondary objectives include correlating environmental exposure to AhR ligands with disease severity in patients Finally we will compare the expression of inflammatory and AhR activation markers in cultured peripheral blood mononuclear cells PBMCs after in vitro stimulation with benzoapyrene
Material and methods
The study will measure exposure to pollutants by determining blood dioxins and urinary PAH metabolites Pro-inflammatory cytokines IL1β TNFα IL23 IL17 and IFNγ and Malondialdehyde MDA serum concentrations will be measured by ELISA The TREG and TH17 lymphocyte population ratio will be evaluated by flow cytometry on isolated PBMCs Additionally the level of expression of CYP 1A1 and 1B1 pollutant-metabolizing enzymes induced by AhR will be assessed on isolated PBMCs The expression levels of the AhR and NfkB active fractions will be determined by immunofluorescence Subsequently levels of AhR activation markers will be compared after stimulation of PBMCs with benzoapyrene
Detailed Description:
I Introduction Pollution is a significant component of the exposome with established links to human health In 2019 pollution was estimated to be responsible for 9 million premature deaths making it the fourth leading risk factor for mortality worldwide Over the last 20 years pollution-related deaths have increased by 66 This is due to industrialization uncontrolled urbanization population growth burning fossil fuels and a lack of restrictive policies in some regions
The cutaneous layer is composed of a complex network of cells that form a mechanical and biological barrier essential for maintaining the bodys integrity and homeostasis Pollution can directly impair this function by crossing the barrier or entering the systemic circulation through inhalation or ingestion
Atopic dermatitis AD and psoriasis are chronic inflammatory skin conditions with multifactorial origins including genetic predisposition immune dysregulation and environmental factors Epidemiological studies have shown a positive correlation between air pollution and the development of these diseases
Given their high prevalence and the impact of symptoms on patients quality of life a better understanding of the exposome in these diseases represents a major challenge
Experimental studies have demonstrated that the aromatic hydrocarbon receptor AhR pathway plays a significant role in the impact of pollutants on psoriasis and AD AhR is a transcription factor involved in the response to environmental pollutants which remains inactive in a cytoplasmic complex with chaperone proteins Upon binding with its ligand AhR translocates to the nucleus and dimerizes with its nuclear partner Arnt The activation of the complex leads to the expression of numerous genes that contain a consensus sequence known as the xenobiotic response element XRE in their promoters
The activator ligands of the AhR comprise xenobiotics found in the environment primarily polycyclic aromatic hydrocarbons PAHs dioxins and so-called dioxin-like compounds 12 This signalling pathway is considered an adaptive response to detoxify the body from various exogenous compounds However prolonged activation of the AhR by such agents may lead to deregulation of the inflammatory process contributing to the development of psoriasis and AD
The aryl hydrocarbon receptor AhR regulates the transcription of cytochromes 1A1 CYP1A1 and 1B1 CYP1B1 which are members of a multigene family of enzymes involved in xenobiotic metabolism These enzymes catalyse the bioactivation of HAPs into electrophilic metabolites leading to activation of NFκB a key player in the inflammatory response During their catalytic cycle these CYP1 enzymes produce reactive oxygen species ROS which generate oxidative stress In addition the AhR is responsible for regulating the expression of pro-inflammatory cytokines including IL1β TNFα IL23 IL17 and IFNγ It can directly affect the balance between Th17 lymphocytes and regulatory T cells Tregs potentially contributing to cutaneous inflammatory processes
However further research is needed to better define the role of AhR in the pathogenesis of chronic inflammatory dermatoses This will help in the management and prevention of these diseases especially when environmental risk factors are associated with predisposition
II Objective
It is hypothesized that the activation of AhR with exogenous ligands may play a role in the development of atopic dermatitis and psoriasis
The main objective is to compare the levels of AhR pathway activation markers between cases and controls
Secondary objectives include correlating environmental exposure to AhR ligands with disease severity in patients
Finally we will compare the expression of inflammatory and AhR activation markers in cultured peripheral blood mononuclear cells PBMCs after in vitro stimulation with benzoapyrene
III Material and methods
We will be carrying out a case-control study
Case Patients with chronic inflammatory dermatosis corresponding to psoriasis or atopic dermatitis
Controls People consulting the dermatology department who do not have these pathologies
All participants including patients and controls will undergo consultationhospitalisation and biological testing as part of their usual medical care Procedures performed as part of the protocol will not interfere with their care
A sample of 10 ml of blood will be taken for toxicological analysis 20 ml of blood will be taken for evaluation of inflammatory and AhR activation markers
A Pollutant measurement The blood sample will be centrifuged the serum will then be collected and stored at -20 C until the blood dioxins are analysed 2378-tétrachlorodibenzo-p-dioxin TCDD and polychlorobiphényle 118 PCB118 will be measured in blood using gas chromatography coupled with high-resolution mass spectrometry GC-HR-MS
For biological monitoring of exposure to PAHs1-Hydroxypyrene 1-OHP will be measured in urine using high-performance liquid chromatography-mass spectrometry LC-MS-MS
B Evaluation of inflammatory and AhR activation markers
PBMCs will be isolated from a 20mL blood sample using a standard Ficoll gradient protocol The isolated cells and sera will be aliquoted and frozen for subsequent experiments on the AhR signalling pathway
Enzyme-Linked Immunosorbent Assay ELISA will be used to measure pro-inflammatory cytokines IL1β TNFα IL23 IL17 and IFNγ
Malondialdehyde MDA concentration will be determined using the ELISA method as a biomarker for lipid peroxidation which indicates the level of oxidative stress caused by reactive oxygen species ROS
Fluorochrome-conjugated monoclonal antibodies CD4 CD25 CD127 FOXP3 Il17 will be used in fluorescence activated cell sorter FACS to assess the numbers and ratios of different lymphocyte populations in PBMCs The investigation will focus on Th17 and Treg levels
FACS will be used to assess the expression of CYP1A1 and CYP1B1 which are prototypes of AhR target genes in PBMCs after specific targeting
The levels of expression of the active fractions of AhR and NfkB will be determined using direct immunofluorescence technique
PBMCs will be cultured with anti-CD3-BaP 50 nM in a cell incubator 37C 5 CO2 controlled humidity for 48-72 hours The parameters of interest including oxidative stress pro-inflammatory cytokines and assessment of the AhR pathway will be reassessed after lymphocyte reactivation by anti-CD3 in the presence or absence of BaP
The cutaneous layer is composed of a complex network of cells that form a mechanical and biological barrier essential for maintaining the bodys integrity and homeostasis Pollution can directly impair this function by crossing the barrier or entering the systemic circulation through inhalation or ingestion
Atopic dermatitis AD and psoriasis are chronic inflammatory skin conditions with multifactorial origins including genetic predisposition immune dysregulation and environmental factors Epidemiological studies have shown a positive correlation between air pollution and the development of these diseases
Given their high prevalence and the impact of symptoms on patients quality of life a better understanding of the exposome in these diseases represents a major challenge
Experimental studies have demonstrated that the aromatic hydrocarbon receptor AhR pathway plays a significant role in the impact of pollutants on psoriasis and AD AhR is a transcription factor involved in the response to environmental pollutants which remains inactive in a cytoplasmic complex with chaperone proteins Upon binding with its ligand AhR translocates to the nucleus and dimerizes with its nuclear partner Arnt The activation of the complex leads to the expression of numerous genes that contain a consensus sequence known as the xenobiotic response element XRE in their promoters
The activator ligands of the AhR comprise xenobiotics found in the environment primarily polycyclic aromatic hydrocarbons PAHs dioxins and so-called dioxin-like compounds 12 This signalling pathway is considered an adaptive response to detoxify the body from various exogenous compounds However prolonged activation of the AhR by such agents may lead to deregulation of the inflammatory process contributing to the development of psoriasis and AD
The aryl hydrocarbon receptor AhR regulates the transcription of cytochromes 1A1 CYP1A1 and 1B1 CYP1B1 which are members of a multigene family of enzymes involved in xenobiotic metabolism These enzymes catalyse the bioactivation of HAPs into electrophilic metabolites leading to activation of NFκB a key player in the inflammatory response During their catalytic cycle these CYP1 enzymes produce reactive oxygen species ROS which generate oxidative stress In addition the AhR is responsible for regulating the expression of pro-inflammatory cytokines including IL1β TNFα IL23 IL17 and IFNγ It can directly affect the balance between Th17 lymphocytes and regulatory T cells Tregs potentially contributing to cutaneous inflammatory processes
However further research is needed to better define the role of AhR in the pathogenesis of chronic inflammatory dermatoses This will help in the management and prevention of these diseases especially when environmental risk factors are associated with predisposition
II Objective
It is hypothesized that the activation of AhR with exogenous ligands may play a role in the development of atopic dermatitis and psoriasis
The main objective is to compare the levels of AhR pathway activation markers between cases and controls
Secondary objectives include correlating environmental exposure to AhR ligands with disease severity in patients
Finally we will compare the expression of inflammatory and AhR activation markers in cultured peripheral blood mononuclear cells PBMCs after in vitro stimulation with benzoapyrene
III Material and methods
We will be carrying out a case-control study
Case Patients with chronic inflammatory dermatosis corresponding to psoriasis or atopic dermatitis
Controls People consulting the dermatology department who do not have these pathologies
All participants including patients and controls will undergo consultationhospitalisation and biological testing as part of their usual medical care Procedures performed as part of the protocol will not interfere with their care
A sample of 10 ml of blood will be taken for toxicological analysis 20 ml of blood will be taken for evaluation of inflammatory and AhR activation markers
A Pollutant measurement The blood sample will be centrifuged the serum will then be collected and stored at -20 C until the blood dioxins are analysed 2378-tétrachlorodibenzo-p-dioxin TCDD and polychlorobiphényle 118 PCB118 will be measured in blood using gas chromatography coupled with high-resolution mass spectrometry GC-HR-MS
For biological monitoring of exposure to PAHs1-Hydroxypyrene 1-OHP will be measured in urine using high-performance liquid chromatography-mass spectrometry LC-MS-MS
B Evaluation of inflammatory and AhR activation markers
PBMCs will be isolated from a 20mL blood sample using a standard Ficoll gradient protocol The isolated cells and sera will be aliquoted and frozen for subsequent experiments on the AhR signalling pathway
Enzyme-Linked Immunosorbent Assay ELISA will be used to measure pro-inflammatory cytokines IL1β TNFα IL23 IL17 and IFNγ
Malondialdehyde MDA concentration will be determined using the ELISA method as a biomarker for lipid peroxidation which indicates the level of oxidative stress caused by reactive oxygen species ROS
Fluorochrome-conjugated monoclonal antibodies CD4 CD25 CD127 FOXP3 Il17 will be used in fluorescence activated cell sorter FACS to assess the numbers and ratios of different lymphocyte populations in PBMCs The investigation will focus on Th17 and Treg levels
FACS will be used to assess the expression of CYP1A1 and CYP1B1 which are prototypes of AhR target genes in PBMCs after specific targeting
The levels of expression of the active fractions of AhR and NfkB will be determined using direct immunofluorescence technique
PBMCs will be cultured with anti-CD3-BaP 50 nM in a cell incubator 37C 5 CO2 controlled humidity for 48-72 hours The parameters of interest including oxidative stress pro-inflammatory cytokines and assessment of the AhR pathway will be reassessed after lymphocyte reactivation by anti-CD3 in the presence or absence of BaP
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None