Ignite Creation Date:
2024-10-26 @ 3:43 PM
Last Modification Date:
2024-10-26 @ 3:43 PM
Study NCT ID:
NCT06655831
Status:
NOT_YET_RECRUITING
Last Update Posted:
None
First Post:
2024-10-21
Brief Title:
Key Mechanisms of Abnormal T Cell Activation and Differentiation in IgG4-Related Ophthalmic Disease
Sponsor:
None
Organization:
None
Study Overview
Official Title:
Study on the Key Mechanisms of Abnormal T Cell Activation and Differentiation in IgG4-Related Ophthalmic Disease
Status:
NOT_YET_RECRUITING
Status Verified Date:
2024-10
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
No
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
IgG4-related disease IgG4-RD is a newly recognized chronic inflammatory condition caused by immune system dysfunction It is characterized by the infiltration of IgG4 plasma cells dense fibrosis and inflammation involving veins and eosinophils Common symptoms include elevated serum IgG4 levels and the formation of tumor-like growths that can affect almost any organ leading to pressure on tissues irreversible damage and even organ failure While estimates suggest a prevalence of 028 to 108 per 100000 people this might be an underestimation due to limited awareness the new definition of the disease and its subtle onset
IgG4-related ophthalmic disease IgG4-ROD is a subtype of IgG4-RD that affects the eye area particularly the lacrimal glands extraocular muscles and surrounding nerves It often presents as painless swelling of the lacrimal glands in one or both eyes sometimes with discomfort or a sensation of a foreign body It can also cause thickening of eye muscles leading to symptoms like bulging eyes blurred vision or double vision In some cases mass lesions in the orbit may press on the optic nerve potentially leading to permanent vision loss While there is currently no cure for IgG4-ROD steroids are used as the main treatment to control inflammation and fibrosis However the disease often recurs with recurrence rates for IgG4-RD reported between 24 and 63 in various studies Understanding the causes of IgG4-ROD could help develop better treatments and reduce the chances of relapse
Studies suggest that T cells play a key role in the development of IgG4-RD including IgG4-ROD CD4 T cells are the main immune cells found in affected tissues They can help B cells multiply and produce IgG4 antibodies and contribute to tissue fibrosis by releasing certain signaling molecules Despite treatment with rituximab a drug that targets B cells many IgG4-RD patients experience relapses indicating that T cells remain important in driving the disease Among the T cell subtypes T follicular helper cells Tfh and CD4 cytotoxic T cells CD4 CTLs are particularly relevant Tfh cells support B cells in producing IgG4 antibodies while CD4 CTLs can contribute to tissue fibrosis by releasing factors like TGF-β IL-1β and IFN-γ However the detailed mechanisms of how T cells become abnormally activated and differentiated in IgG4-ROD remain unclear
This study will use samples from the lacrimal glands blood and tears of IgG4-ROD patients to investigate how T cells become abnormally active and differentiate in this condition The findings could identify new targets for therapy helping to reduce the recurrence of IgG4-ROD and provide insights into treating other forms of IgG4-RD
IgG4-related ophthalmic disease IgG4-ROD is a subtype of IgG4-RD that affects the eye area particularly the lacrimal glands extraocular muscles and surrounding nerves It often presents as painless swelling of the lacrimal glands in one or both eyes sometimes with discomfort or a sensation of a foreign body It can also cause thickening of eye muscles leading to symptoms like bulging eyes blurred vision or double vision In some cases mass lesions in the orbit may press on the optic nerve potentially leading to permanent vision loss While there is currently no cure for IgG4-ROD steroids are used as the main treatment to control inflammation and fibrosis However the disease often recurs with recurrence rates for IgG4-RD reported between 24 and 63 in various studies Understanding the causes of IgG4-ROD could help develop better treatments and reduce the chances of relapse
Studies suggest that T cells play a key role in the development of IgG4-RD including IgG4-ROD CD4 T cells are the main immune cells found in affected tissues They can help B cells multiply and produce IgG4 antibodies and contribute to tissue fibrosis by releasing certain signaling molecules Despite treatment with rituximab a drug that targets B cells many IgG4-RD patients experience relapses indicating that T cells remain important in driving the disease Among the T cell subtypes T follicular helper cells Tfh and CD4 cytotoxic T cells CD4 CTLs are particularly relevant Tfh cells support B cells in producing IgG4 antibodies while CD4 CTLs can contribute to tissue fibrosis by releasing factors like TGF-β IL-1β and IFN-γ However the detailed mechanisms of how T cells become abnormally activated and differentiated in IgG4-ROD remain unclear
This study will use samples from the lacrimal glands blood and tears of IgG4-ROD patients to investigate how T cells become abnormally active and differentiate in this condition The findings could identify new targets for therapy helping to reduce the recurrence of IgG4-ROD and provide insights into treating other forms of IgG4-RD
Detailed Description:
Immunoglobulin-G4 related disease IgG4-RD is a newly defined immune-mediated chronic inflammatory disease with fibrosis characterized by lymphoplasmacytic infiltration predominantly consisting of IgG4 plasma cells along with storiform fibrosis obliterative phlebitis and eosinophil infiltration The most common clinical manifestations of IgG4-RD include significantly elevated serum IgG4 levels and mass-like lesions which can affect nearly any part of the body leading to compression and irreversible damage to affected organs and surrounding tissues potentially resulting in organ failure Despite reports estimating the prevalence of IgG4-RD to be between 028108100000 the actual prevalence might be higher due to its recent definition limited awareness among clinicians and patients and its insidious onset
Immunoglobulin-G4 related ophthalmic disease IgG4-ROD refers to IgG4-RD mass-like lesions involving the lacrimal gland extraocular muscles supraorbital nerve infraorbital nerve and other orbital structures leading to orbital lesions Among these lacrimal gland involvement is the most common manifesting as painless unilateral or bilateral swelling sometimes accompanied by foreign body sensation or eye discomfort Extraocular muscle involvement can present as muscle enlargement potentially causing proptosis blurred vision and diplopia in severe cases Orbital mass lesions may also compress the optic nerve leading to irreversible vision loss While there is currently no cure for IgG4-ROD glucocorticoids are used as the first-line treatment to control inflammation and fibrosis associated with the mass-like lesions IgG4-ROD is prone to relapse and though there are no studies specifically on its recurrence rate different studies have reported relapse rates of IgG4-RD patients during follow-up ranging from 24 to 63 The exact etiology and pathogenesis of IgG4-ROD remain unclear and further elucidation of these aspects would facilitate the identification of new therapeutic targets reduce relapse rates and potentially achieve a cure
Multiple studies on various forms of IgG4-RD suggest a critical role of T cells in the pathogenesis of IgG4-RD including IgG4-ROD First CD4 T cells constitute the majority of infiltrating lymphocytes in the lesions of IgG4-RD These CD4 T cells induce B cell proliferation and class-switching through cytokine secretion and contribute to tissue fibrosis Additionally IgG4-RD patients treated with rituximab exhibit high relapse rates with post-relapse B cell clones expanding differently compared to pre-treatment This evidence strongly supports the central role of T cells in IgG4-RD Among the T cell subpopulations implicated in IgG4-RD pathogenesis T follicular helper cells Tfh and CD4 cytotoxic T lymphocytes CD4 CTLs are particularly notable Tfh cells promote ectopic germinal center formation and B cell class-switching to IgG4 plasma cells through IL-21 secretion while CD4 CTLs contribute to tissue fibrosis by secreting TGF-β IL-1β and IFN-γ However the mechanisms underlying abnormal T cell activation and differentiation in both IgG4-ROD and other IgG4-RD remain poorly understood
This study aims to investigate key pathways involved in abnormal T cell activation and differentiation in IgG4-ROD by conducting a series of in vitro experiments using surgical tissue samples peripheral blood and tear fluid from patients with IgG4-ROD These experiments include RNA sequencing RNA-Seq enzyme-linked immunosorbent assays ELISA and in vitro cell co-culture The goal is to identify critical pathways in the pathogenesis of IgG4-ROD and to clarify their roles in the disease process providing new therapeutic targets to reduce the recurrence rate of IgG4-ROD This research may also offer insights into the mechanisms and treatment of other IgG4-related diseases
Study Population
This study involves patients from the Department of Ophthalmology at Peking University Third Hospital including lacrimal gland tissue samples from IgG4-ROD patients experimental group and patients with lacrimal gland ptosis undergoing surgical repositioning control group The inclusion and exclusion criteria are as follows
Inclusion Criteria
Experimental Group IgG4-ROD patients with lacrimal gland involvement diagnosed based on 2014 and 2020 Japanese criteria for IgG4-ROD and IgG4-RD
Clinical and imaging features characteristic diffuse or localized swelling or nodular formations of the lacrimal glands
Serological diagnosis Elevated serum IgG4 concentration 135 mgdL
Histopathological diagnosis meeting 2 of the 3 criteria below
Dense lymphoplasmacytic infiltration with fibrosis IgG4 plasma cellIgG plasma cell ratio 40 and 10 IgG4 plasma cells per high-power field
Characteristic fibrosis especially storiform fibrosis or obliterative phlebitis
Control Group Patients with lacrimal gland ptosis confirmed by pathology and clinicalimaging features requiring surgical repositioning
Exclusion Criteria
Presence of other autoimmune diseases like Sjögrens syndrome systemic lupus erythematosus or rheumatoid arthritis
Lacrimal gland pathology indicating other orbital diseases such as MALT lymphoma or inflammatory pseudotumor
Other exclusion criteria include mental disorders malignancies or any condition deemed unsuitable for study participation
Study Methods
Collection of Clinical Information Patient demographics medical history and necessary diagnostic test results eg imaging serum IgG levels are recorded Ocular examinations include visual acuity intraocular pressure Ocular Surface Disease Index OSDI corneal fluorescein staining CFS tear film break-up time FBUT and Schirmers test among others
Collection of Samples Tear fluid about 15 µL per eye is collected using a capillary tube and stored at -80C A portion 50-100 mg of the resected lacrimal gland tissue is reserved for this study while the rest is used for pathological analysis Blood samples approximately 10 mL are drawn for preparing peripheral blood mononuclear cells PBMCs
Bulk RNA-Seq Analysis of Lacrimal Gland Tissue Total RNA is extracted from lacrimal gland tissue using the Trizol method followed by RNA quality assessment with an Agilent bioanalyzer A cDNA library is constructed and sequenced using Illumina technology Data processing includes quality control with fastp alignment using HISAT2 and gene expression quantification with featureCounts Differential expression analysis is performed using DESeq2 and Gene Set Enrichment Analysis GSEA is conducted Immune cell composition in the tissue is analyzed using xCell and CIBERSORTx
Enzyme-linked Immunosorbent Assay ELISA ELISA is used to analyze key pathways identified in the RNA-Seq results using specific kits according to the manufacturers instructions
In Vitro Co-culture Experiments An in vitro co-culture system of IgG4-ROD patient lacrimal gland cells and PBMCs is established to simulate the immune microenvironment of the lacrimal gland Based on RNA-Seq and ELISA results targeted interventions are applied to this system IgG4IgG1 ratios in the supernatant serve as an indicator with higher values suggesting stronger pathogenic effects
Immunoglobulin-G4 related ophthalmic disease IgG4-ROD refers to IgG4-RD mass-like lesions involving the lacrimal gland extraocular muscles supraorbital nerve infraorbital nerve and other orbital structures leading to orbital lesions Among these lacrimal gland involvement is the most common manifesting as painless unilateral or bilateral swelling sometimes accompanied by foreign body sensation or eye discomfort Extraocular muscle involvement can present as muscle enlargement potentially causing proptosis blurred vision and diplopia in severe cases Orbital mass lesions may also compress the optic nerve leading to irreversible vision loss While there is currently no cure for IgG4-ROD glucocorticoids are used as the first-line treatment to control inflammation and fibrosis associated with the mass-like lesions IgG4-ROD is prone to relapse and though there are no studies specifically on its recurrence rate different studies have reported relapse rates of IgG4-RD patients during follow-up ranging from 24 to 63 The exact etiology and pathogenesis of IgG4-ROD remain unclear and further elucidation of these aspects would facilitate the identification of new therapeutic targets reduce relapse rates and potentially achieve a cure
Multiple studies on various forms of IgG4-RD suggest a critical role of T cells in the pathogenesis of IgG4-RD including IgG4-ROD First CD4 T cells constitute the majority of infiltrating lymphocytes in the lesions of IgG4-RD These CD4 T cells induce B cell proliferation and class-switching through cytokine secretion and contribute to tissue fibrosis Additionally IgG4-RD patients treated with rituximab exhibit high relapse rates with post-relapse B cell clones expanding differently compared to pre-treatment This evidence strongly supports the central role of T cells in IgG4-RD Among the T cell subpopulations implicated in IgG4-RD pathogenesis T follicular helper cells Tfh and CD4 cytotoxic T lymphocytes CD4 CTLs are particularly notable Tfh cells promote ectopic germinal center formation and B cell class-switching to IgG4 plasma cells through IL-21 secretion while CD4 CTLs contribute to tissue fibrosis by secreting TGF-β IL-1β and IFN-γ However the mechanisms underlying abnormal T cell activation and differentiation in both IgG4-ROD and other IgG4-RD remain poorly understood
This study aims to investigate key pathways involved in abnormal T cell activation and differentiation in IgG4-ROD by conducting a series of in vitro experiments using surgical tissue samples peripheral blood and tear fluid from patients with IgG4-ROD These experiments include RNA sequencing RNA-Seq enzyme-linked immunosorbent assays ELISA and in vitro cell co-culture The goal is to identify critical pathways in the pathogenesis of IgG4-ROD and to clarify their roles in the disease process providing new therapeutic targets to reduce the recurrence rate of IgG4-ROD This research may also offer insights into the mechanisms and treatment of other IgG4-related diseases
Study Population
This study involves patients from the Department of Ophthalmology at Peking University Third Hospital including lacrimal gland tissue samples from IgG4-ROD patients experimental group and patients with lacrimal gland ptosis undergoing surgical repositioning control group The inclusion and exclusion criteria are as follows
Inclusion Criteria
Experimental Group IgG4-ROD patients with lacrimal gland involvement diagnosed based on 2014 and 2020 Japanese criteria for IgG4-ROD and IgG4-RD
Clinical and imaging features characteristic diffuse or localized swelling or nodular formations of the lacrimal glands
Serological diagnosis Elevated serum IgG4 concentration 135 mgdL
Histopathological diagnosis meeting 2 of the 3 criteria below
Dense lymphoplasmacytic infiltration with fibrosis IgG4 plasma cellIgG plasma cell ratio 40 and 10 IgG4 plasma cells per high-power field
Characteristic fibrosis especially storiform fibrosis or obliterative phlebitis
Control Group Patients with lacrimal gland ptosis confirmed by pathology and clinicalimaging features requiring surgical repositioning
Exclusion Criteria
Presence of other autoimmune diseases like Sjögrens syndrome systemic lupus erythematosus or rheumatoid arthritis
Lacrimal gland pathology indicating other orbital diseases such as MALT lymphoma or inflammatory pseudotumor
Other exclusion criteria include mental disorders malignancies or any condition deemed unsuitable for study participation
Study Methods
Collection of Clinical Information Patient demographics medical history and necessary diagnostic test results eg imaging serum IgG levels are recorded Ocular examinations include visual acuity intraocular pressure Ocular Surface Disease Index OSDI corneal fluorescein staining CFS tear film break-up time FBUT and Schirmers test among others
Collection of Samples Tear fluid about 15 µL per eye is collected using a capillary tube and stored at -80C A portion 50-100 mg of the resected lacrimal gland tissue is reserved for this study while the rest is used for pathological analysis Blood samples approximately 10 mL are drawn for preparing peripheral blood mononuclear cells PBMCs
Bulk RNA-Seq Analysis of Lacrimal Gland Tissue Total RNA is extracted from lacrimal gland tissue using the Trizol method followed by RNA quality assessment with an Agilent bioanalyzer A cDNA library is constructed and sequenced using Illumina technology Data processing includes quality control with fastp alignment using HISAT2 and gene expression quantification with featureCounts Differential expression analysis is performed using DESeq2 and Gene Set Enrichment Analysis GSEA is conducted Immune cell composition in the tissue is analyzed using xCell and CIBERSORTx
Enzyme-linked Immunosorbent Assay ELISA ELISA is used to analyze key pathways identified in the RNA-Seq results using specific kits according to the manufacturers instructions
In Vitro Co-culture Experiments An in vitro co-culture system of IgG4-ROD patient lacrimal gland cells and PBMCs is established to simulate the immune microenvironment of the lacrimal gland Based on RNA-Seq and ELISA results targeted interventions are applied to this system IgG4IgG1 ratios in the supernatant serve as an indicator with higher values suggesting stronger pathogenic effects
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
None
Is a FDA Regulated Device?:
None
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None